CHAPTER-2

LIPIDS

The lipids (in Greek lipos means-fat) are a heterogeneous group of organic compounds found in
cells. Most common and defining feature of lipids are their insolubility in water, but they
dissolve in polar solvents such as ether, alcohol, chloroform and benzene. They combine with
carbohydrates and proteins to form macronutrients. The lipids form about 3.5% of cell contents.

FATTY ACIDS:

These are carboxylic acids with a long hydrocarbon chain, ranging from 4-36 carbons long, the
chains are usually linear, saturated (When only single bonds exist and no double bonds are
present then it is said to be saturated) or unsaturated acids (With one or more double bonds
(C=C) the fatty acid is said to be unsaturated). Fatty acids are obtained from the hydrolysis of
fats.

Classification Of Fatty Acids:

They can be functionally classified based on the length of the chain as follows:
• Short-chain fatty acids: up to 6 carbon atoms
• Medium-chain fatty acids: 8 to 12 carbon atoms.
• Long-chain fatty acids: 14 to 18 carbon atoms
• Very long-chain fatty acids: 20 carbon atoms and up.

Fatty acids can be divided into two broad classes based on the presence or absence of double
bonds:
Saturated fatty acids: If there are no double bonds in the carbon chain or if there are only single
bonds between neighboring carbons in the hydrocarbon chain, it is saturated. Stearic acid and
palmitic acid, which are commonly found in meat, are examples of saturated fats.
Unsaturated fatty acids: If there are one or more double bonds in the carbon chain, it is
unsaturated. Oleic acid is an example of an unsaturated fatty acid.
• Each isomeric structure causes the unsaturated fats to exhibit different properties due to
the location of its bonds and, therefore, its energy configuration. The reason that trans
fatty acids are more often solid, while cis are often liquid, is due to their structures.
• Cis fatty acids have a geometric shape which causes the hydrogen atoms to be on the
same side of the molecule.
• Trans fatty acids have a geometric shape in which the hydrogen atoms are on opposite
sides of the molecule.

Trans fatty acids:

• Trans fats are a form of unsaturated fat and come in both natural and artificial forms. Or
we can say that two broad types of trans fats found in foods, namely naturally occurring
and artificial trans fats.
• Trans fats are the most harmful type of fats which can cause an adverse effect on a
human body than any other dietary constituent.
• Naturally-occurring trans fats are produced in the gut of some animals and foods made
from these animals example milk and meat products.
• They may contain small quantities of these fats. Artificial trans fats are generated through
an industrial process that adds hydrogen to liquid vegetable oils to form them more solid.
• That is hydrogen is made to react with the oil to produce fats that resemble pure ghee or
butter.
• Let us tell you that the primary source for trans fats in processed food is "partially
hydrogenated oils".
 • Trans Fat in Food Partially hydrogenated oil is the manufactured form of trans fat that
may be found in various variety of food products like; - Baked goods including cakes,
cookies, and pies. - Microwave popcorn - Shortening - Frozen Pizza - Refrigerated dough
like biscuits and rolls - Nondairy coffee creamer - Fried Foods like french fries,
doughnuts, and fried chicken - Stick margarine.
• Therefore, we can say that trans fatty acid contains oils that can be preserved longer, food
can be transformed into the desired shape and texture, and can easily substitute 'Pure
Ghee'.

The harmful effects of Trans fatty acids:

• TFA increases the risk of heart attacks, stroke, and type 2 diabetes.
• They also pose an unhealthy effect on cholesterol levels. That is they not only increase
total cholesterol levels but also reduce good cholesterol (HDL), which helps to protect
against heart disease.
• It also increases the risk of developing obesity, metabolic syndrome, insulin resistance,
infertility, certain types of cancers and can also lead to compromised fetal development
that can cause harm to the yet to be born baby.
• Consumption of trans-unsaturated fatty acids has shown to increase the risk of ovulatory
infertility in women.
• Trans fatty acids promote systemic inflammation by increasing the levels of c-reactive
protein(CRP) leading to thickening of arteries (atherosclerosis), diabetes, sudden death
due to heart failure.

MUFA:
• Monounsaturated fatty acids (MUFA) are chemically classified as fatty acids containing a
single double bond.
• They are synthesized endogenously in humans, though not fully compensating the body
requirements. So it is considered as partially essential fatty acids.
• These fats are usually liquid when at room temperature and turn solid when chilled.
• MUFA is considered as healthy fats since they can lower the blood cholesterol level and
reduce the risk of cardiovascular diseases and strokes.
• MUFA are rich in vitamin E, which is an antioxidant, which is necessary to keep our
body healthy by protecting cell damages.
• Although animal products such as eggs and meat contain MUFAs, the richest sources are
plant foods.
• Good sources of MUFAs include: olive oil, canola oil, peanut oil, sesame oil, sunflower
oil, avocados, peanut butter, nuts and seeds.
• The most common cis-configured MUFA in daily nutrition is oleic acid and palmitoleic
acid (16:1 n-7).
Clinical Significance:
• Cardiovascular health: MUFAs can help lower LDL (bad) cholesterol levels, which can
reduce the risk of heart disease and stroke.
• MUFA play a significant role in cholesterol absorption.
• MUFAs help prevent cancer and diabetes.
• MUFAs help prevent LDL oxidation and thus atherosclerosis.
• High MUFA content in diet acts as the key to beneficial effects on joints.

Polyunsaturated Fatty Acids (PUFA):

• Polyunsaturated fatty acids (PUFA) are healthy lipid molecules which have two or more
carbon-carbon double bonds.
• They are never conjugated PUFAs have two or more cis double bonds that are separated
from each other by a single methylene group.
• These double bonds generally occur after every three carbon atoms.
• Oils rich in PUFA are liquid in room temperature.
• The human body cannot synthesize PUFAs, so they must be obtained from food.
• Good sources of PUFA are walnut, sunflower seeds, flaxseeds and poppy seeds. PUFA
are of two types omega 3 fatty acids and omega 6 fatty acids.
• Omega 3 fatty acids are of 3 types linoleic acid, Eicosapentaenoic Acid (EPA) and
Docosahexaenoic Acid (DHA).
• It is abundant mostly in fish and some of vegetarian sources are chia seeds, hemp seeds
and flaxseeds.
• The omega-6 fatty acid, linoleic-acid, and arachidonic acid.

Biological significance:
• These are important for growth and normal health
• They lower blood cholesterol level
• They are important constituents of plasma membrane
• They are important for transport and metabolism of cholesterol
• When these are taken through diet, they reduce the risk for heart disease
• They are important for nerve function, blood clotting, brain health and muscle strength.
• Their deficiency leads to growth retardation, loss of hair, dermatitis and poor wound
healing.
DIGESTION OF LIPIDS:

Lipid digestion can be broken into three main steps:

• Lipid digestion in the mouth
• Lipid digestion in the stomach
• Lipid digestion in the small intestine

Both the mouth and stomach play a role in lipid digestion, but the majority of lipid digestion
occurs in the small intestine. Once lipids are broken down through these steps. Triglycerides are
large molecules, and they’re not water-soluble. Because of this, they like to cluster together in
large droplets when they’re in a watery environment like the digestive tract. The digestive
process has to break those large droplets of fat into smaller droplets and then enzymatically
digest lipid molecules using enzymes called lipases.

Chewing mechanically breaks food into smaller particles and mixes them with saliva. An
enzyme called lingual lipase is produced by cells on the tongue (“lingual” means relating to the
tongue) and begins some enzymatic digestion of triglycerides, cleaving individual fatty acids into
small droplets. In the stomach, mixing and churning helps to disperse food particles and fat
molecules. Cells in the stomach produce another lipase, called gastric lipase (“gastric” means
relating to the stomach) that also contributes to enzymatic digestion of triglycerides. Lingual
lipase swallowed with food and saliva also remains active in the stomach. As the stomach
contents enter the small intestine, most of the dietary lipids are undigested and clustered in large
droplets. Secretion of pancreatic juice and bile, which is made in the liver and stored in the
gallbladder, is released into the duodenum, (which is the first section of the small intestine) by
the contractions of smooth muscles of gall bladder. Bile contains no enzyme but water, mucin,
lecithin and bile salts. Bile salts have both a hydrophobic and a hydrophilic side, so they are
attracted to both fats and water. This makes them effective emulsifiers, meaning that they break
large fat globules into smaller droplets. Emulsification makes lipids more accessible to digestive
enzymes by increasing the surface area for them to act.
The pancreas secretes pancreatic lipase into the small intestine to enzymatically digest
triglycerides. Triglycerides are broken down to fatty acids, monoglycerides (glycerol backbone
with one fatty acid still attached), and some free glycerol. Cholesterol and fat-soluble vitamins
do not need to be enzymatically digested.

Triacylglycerol pancreatic lipase 3 fatty acids + Glycerol Triglyceride hydrolysis releases free
fatty acids (FFAs) and monoacylglycerols. Phospholipid hydrolysis yields FFAs and
lysophospholipids. Cholesterol esters are hydrolyzed to free cholesterol and FFAs. In the small
intestine besides pancreatic secretion, fatty food is also acted upon by intestinal lipolytic
enzymes intestinal lipase of intestinal juice. It hydrolyzes triglycerides, diglycerides and
monoglycerides to fatty acids and glycerol. Triglyceride intestinal lipase monoglyceride +fatty
acids

ABSORPTION OF FATS:

The products of fat digestion (fatty acids, monoglycerides, glycerol, cholesterol) need to enter
into the circulation so that they can be used by cells around the body. Again, bile helps with this
process. Bile salts cluster around the products of fat digestion to form structures called micelles,
which help the fats get close enough to the microvilli of intestinal cells so that they can be
absorbed. The products of fat digestion diffuse across the membrane of the intestinal cells, and
bile salts are recycled back to do more work emulsifying fat and forming micelles.
Emulsification is important for the digestion of lipids because lipases can only efficiently act on
the lipids when they are broken into small aggregates.

Once inside the intestinal cell, short- and medium-chain fatty acids and glycerol can be directly
absorbed into the bloodstream rapidly, through simple diffusion but larger lipids such as long-
chain fatty acids, are absorbed slowly. Over 98% of dietary lipids is normally absorbed.
Cholesterol is also absorbed in the free form.
COMPOUNDS FORMED FROM CHOLESTEROL:
There are two classes of important molecules are dericed from cholesterol, they are
1. Bile acids
2. Steroid hormones

Bile acids:
• Bile acids and bile salts are detergents that emulsify the fats in the gut during digestion.
These are synthesized from cholesterol in the liver. The bile acids has 4 rings (A,B,C,D) and
they undergo series of reactions they are
• Addition of hydroxyl group into ring B and ring C
• Shorten the acyl chain to 7 chain with last carbon changing to carboxyl group of ring D
• Taurine and glycine are linked to carboxyl group (it’s a side chain) resulting in bile salts
called cholic acid and chenodeoxycholic acid.
• Bile salts lack hydroxyl group on ring c.
• These bile acids are secreted from the liver into the gall bladder for storage, when there is
requirement it is released into the small intestine through bile duct.
• After the doing emulsification action in fat digestion, they are reabsorbed into the blood and
goes to liver and gets reused. This cycle is called enterohepatic circulation.
• This circulation handles around 20-30g of bile acids/day in human beings. If small fraction
escape this cycle they are lost in feces.

Steroid hormones:
They consume a very small fraction of total cholesterol available in the organism. There are five
principal classes, all derived from cholesterol
1. progestin’s (active during pregnancy)
2. glucocorticoids (promotes the synthesis of glucose and suppresses inflammatory
reactions)
3. mineralocorticoids(regulating ion balance)
4. androgens(promoting male sex characteristics)
5. estrogens(promoting female sex charcateristics)

Except for progesterone all the other hormones have shortened side chain in ring D.
In some cases they have oxidized OH group in ring A.
Each and every hormones are synthesized on demand by the pregnant women, adrenal cortex by
the gonads.
ATHEROSCLEROSIS:

Atherosclerosis is the thickening or hardening of the arteries due to the buildup of fats,
cholesterol, calcium, and fibrin and other substances in and on the artery walls. This buildup is
called plaque. The plaque can cause arteries to narrow, blocking blood flow. As it builds up in
the arteries, the artery walls become thickened and stiff. This reduces blood flow and oxygen
supply to the vital body organs and extremities the plaque can also burst, leading to a blood clot.
Although atherosclerosis is often considered a heart problem, it can affect arteries anywhere in
the body. Atherosclerosis can be treated. Healthy lifestyle habits can help prevent
atherosclerosis.

Risk factors for atherosclerosis, include:

• High cholesterol and triglyceride levels
• High blood pressure
• Smoking
• Type 1 diabetes
• Obesity
• Physical inactivity
• High saturated fat diet

Signs and symptoms:

Atherosclerosis may develop gradually, and may be few, as the plaque gradually builds up in the
artery. However, when a major artery is blocked, signs and symptoms may be severe, such as
those occurring with heart attack, stroke, or blood clot. Once symptoms develop, these may
include:
• chest pain(stable angina)
• pain in your leg, arm, and other regions supplied by the blocked artery
• cramping in the buttocks while walking
• heart-palpitations
• shortness of breath
• fatigue
 • mental confusion if the blockage affects
• cerebral-circulation
• weakness and loss of sensation on one side of the body if the blockage affects brain
circulation
• muscle weakness and cramps in your legs from lack of circulation
• Vision and speech problem.

Diagnostics:
Test that evaluate the atherosclerotic arteries are:
1. Angiography
2. Cardiac catheterization
3. Ultrasound
4. Electrocardiogram (ECG)

LIPOPROTEINS:

• Lipoproteins are complex particles that have a central hydrophobic core and a
hydrophilic membrane
• The core is comprised of non-polar lipids (like cholesterol esters and triglycerides)
• The membrane contains phospholipids, free cholesterol and apolipoproteins.
• All these components are insoluble in water (like cholesterol esters, cholesterol,
phospholipids, triglycerides), and hence they have to be transported from the tissue of
origin too tissues to which they are stored or consumed.
• These are carried in the blood called as plasma lipoproteins. The specific carriers of
lipoproteins are apolipoproteins.
• These apolipoproteins are divided into different classes from A to E.

Types of lipoproteins:
Chylomicrons:
these are the largest and least dense of the lipoproteins, with the highest triglyceride content.
They consist of a protein component synthesized in the liver, and transport the dietary
triglycerides and cholesterol to different tissues. The diameter of chylomicrons ranges from
about 100-500nm. The apolipoproteins present are A-1,A-2,A-4,A-5,B-48,C-2,C-3 and E. apo B-
48 is the core structural protein and each chylomicron will have one apo B-48. It travels to the
liver, where the cholesterol is metabolized. Thus chylomicrons deliver fats and cholesterol from
the intestines to the muscles, fat cells and the liver. whereas in the fasting state the chylomicron
particles are small carrying decreased quantities of triglyceride.

Chylomicron Remnants:
The removal of triglyceride from chylomicrons by peripheral tissues results in smaller particles
called chylomicron remnants. Compared to chylomicrons these particles are enriched in
cholesterol and are pro-atherogenic.

Very Low-Density Lipoproteins (VLDL):

Very low density lipoproteins are large particles of diameter in range, of 25-70 nm. These
particles are produced by the liver and are triglyceride rich. The core of VLDL consist of 50-
60% triacylglycerols and the surface layer contains cholesterol, cholesteryl esters, phospholipids
and apoproteins apo B-100, apo C-I, apo C-II, apo C-III and apo-E. Apo B-100 is the core
structural protein and each VLDL particle contains one Apo B-100 molecule. Similar to
chylomicrons, the size of the VLDL particles can vary depending on the quantity of triglyceride
carried in the particle. When triglyceride production in the liver is increased, the secreted VLDL
particles are large. However, VLDL particles are smaller than chylomicrons.

Intermediate-Density Lipoproteins (IDL; VLDL Remnants):

The removal of triglycerides from VLDL by muscle and adipose tissue results in the formation
of IDL particles which are enriched in cholesterol. These particles contain apolipoprotein B-100
and E. These IDL particles are pro-atherogenic.

Low-Density Lipoproteins (LDL):

Low density lipoproteins are small particles of diameter 20 to 25 nm. LDL is also called as bad
cholesterol. These particles are derived from VLDL and IDL particles and they are even further
enriched in cholesterol. LDL carries the majority of the cholesterol that is in the circulation. The
predominant apolipoprotein is B-100 and each LDL particle contains one Apo B-100 molecule
(see figure 2.34 C). An abundance of small dense LDL particles are seen in association with
hypertriglyceridemia, low HDL levels, obesity, type 2 diabetes and infectious and inflammatory
states. These small dense LDL particles are considered to be more pro-atherogenic than large
LDL particles. Small dense LDL particles have a decreased affinity for the LDL receptor
resulting in a prolonged retention time in the circulation. Additionally, they more easily enter the
arterial wall and bind more avidly to intra-arterial proteoglycans, which traps them in the arterial
wall.

High-Density Lipoproteins (HDL):

High Density Lipoproteins are small, protein rich particles of diameter in the range of 8 to 11
nm. These lipoproteins contain relatively little cholesterol and no cholesteryl esters. HDLs
contain apo A-I, apo C-I, apo C-Il and other apolipoproteins. Apo A-I is the core structural
protein and each HDL particle may contain multiple Apo A-I molecules. HDL particles are
enriched in cholesterol and phospholipids. These particles play an important role in reverse
cholesterol transport from peripheral tissues to the liver, which is one potential mechanism by
which HDL may be anti-atherogenic. In addition, HDL particles have anti-oxidant, anti-
inflammatory, anti-thrombotic, and anti-apoptotic properties, which may also contribute to their
ability to inhibit atherosclerosis.

Functions of Lipoproteins:
1. Lipoproteins play a key role in the absorption and transport of dietary lipids by the small
intestine, in the transport of lipids from the liver to peripheral tissues, and the transport of lipids
from peripheral tissues to the liver and intestine (reverse cholesterol transport).
2. High-density lipoprotein (HDL) is the "good cholesterol". It carries cholesterol back to your
liver to be flushed out of your body. High levels of HDL reduce your risk of cardiovascular
(heart) disease.
3. Low-density lipoprotein (LDL) is the "bad cholesterol". It increases your risk of coronary
artery disease, heart attacks, and strokes. LDL carries cholesterol that accumulates as plaque
inside blood vessels. Plaque buildup can make blood vessels too narrow for blood to flow freely.
This condition is atherosclerosis.
4. Very low-density lipoproteins (VLDL) are another type of "bad cholesterol". VLDLs carry
triglycerides and to a less degree, cholesterol, to your tissues.
5. Intermediate-density lipoproteins (IDL) are created as VLDLs give up their fatty acids. They
are then either removed by the liver or converted into LDL.
6. Chylomicrons are very large particles that also transport triglycerides.

KETONE BODIES AND ITS TYPES

Ketone bodies are three water-soluble compounds that are produced as by-products when fatty
acids are broken down for energy in the liver and kidney. The term 'ketone bodies' refers to three
molecules,
1. acetoacetate (AcAc),
2. 3-ß-hydroxybutyrate (3HB)
3. acetone.
Two of the three are used as a source of energy in the heart and brain while the third is a waste
product excreted from the body. In humans and most other mammals, acetyl CoA formed in the
liver during oxidation of fatty acids can either enter the citric acid cycle or may undergo
conversion to the ketone bodies. Acetoacetate accumulates during fatty acid metabolism under
low carbohydrate conditions. 3-ß-hydroxybutyrate is formed from the reduction of AcAc in the
mitochondria. These two predominant ketone bodies are energy-rich compounds that transport
energy from the liver to other tissues. Acetone is generated by spontaneous decarboxylation of
AcAc and is responsible for the sweet odor on the breath of individuals with ketoacidosis.
Acetone is produced in smaller quantities than other ketone bodies. During glucose deficiency,
ketone bodies play a key role in sparing glucose utilization and reducing proteolysis. Unlike
most other tissues, the brain cannot utilize fatty acids for energy when blood glucose levels
become compromised. In this case, ketone bodies provide the brain with an alternative source of
energy, amounting to nearly 2/3 of the brain's energy needs during periods of prolonged fasting
and starvation.

Clinical Significance of ketone bodies:

An overproduction of ketone bodies through increased ketogenesis can pose a problem due to
their acidic nature. The most prevalent subdivisions of ketoacidosis are diabetic ketoacidosis
(DKA) and alcoholic ketoacidosis. DKA most commonly occurs as a result of a lack of
appropriate response to insulin in the body. This lack of insulin response could be due to non-
compliance with treatment, undiagnosed type 1 DM, or sub- therapeutic administration of
insulin. A lack of insulin response results in an increase of glucagon and a decrease in the uptake
of glucose. Decreased insulin also facilitates the increased activity of hormone- sensitive lipase.
Hormone-sensitive lipase converts triglycerides to fatty acyl CoA molecules.
Fatty acyl CoA molecules overload the Krebs cycle and shunt over to ketone metabolism. An
abundance of ketone bodies causes an anion gap acidosis. Ultimately, the high glucose levels
lead to osmotic diuresis, involving greater osmole concentrations that cause an increased osmotic
pressure, which leads to reduced water reabsorption in the kidneys. Along with being
dehydrated, patients typically present with confusion, nausea, vomiting, and abdominal pain.
Because of the acidosis, patients often breathe very deeply and rapidly to eliminate carbon
dioxide and cause respiratory alkalosis. This process is known as Kussmaul's breathing, and,
over time, a patient experiences respiratory distress due to the prolonged exertion of respiratory
muscles. Cerebral edema occurs in severe cases of DKA. Because of the acetone produced by
ketogenesis, the breath of patients smells fruity or like nail polish remover. Ketoacidosis also can
occur with severe alcoholism and prolonged starvation.
RELATED DISORDERS OF LIPID METABOLISM

1. Steatorrhea:
Among the macronutrients, digestion and absorption of fat involve a complex mechanism. Fat
absorption requires bile acids, digestive enzymes, and a normally functioning small intestinal
mucosa. Dietary lipids, mostly as triacylglycerols, are initially emulsified by bile acids and then
hydrolyzed by the pancreatic lipases and colipases into free fatty acids and monoglycerides. In
the proximal small bowel, these hydrolyzed lipids form micelles by the action of bile acids. The
micelles are then absorbed across the intestinal villi and transported as chylomicrons via the
intestinal lymphatics. Due to the biliary obstruction or pancreatic disease, person fails to digest
or absorb fat that results its excretion in feces in large amounts and leads to a disease condition
called steatorrhea. Steatorrhea is one of the clinical features of fat malabsorption and noted in
many conditions such as exocrine pancreatic insufficiency (EPI), celiac disease, and tropical
sprue.
Symptoms associated with severe or chronic steatorrhea include chronic loose, heavy, foul-
smelling fat-filled stool, anemia, muscle weakness and pain, chronic exhaustion, weight loss,
fever, reduced growth rate in children, vision problems, skin conditions, neurological conditions
and osteoporosis.
2. Gall stones (Cholelithiasis):
Gallstone disease (GSD) is a common biliary disorder in which abnormal regulation of lipid
metabolism is observed through hypersecretion of hepatic cholesterol into bile with less frequent
hyposecretion of bile salts or phospholipids. Normally, bile contains enough chemicals to
dissolve the cholesterol excreted by liver. But if liver excretes more cholesterol than bile can
dissolve, the excess cholesterol may combine with bile salts to form into crystals which
eventually develop into stones. Complications of gall stones include: inflammation of the gall
bladder (cholecystitis) which can cause severe pain and fever, blockage of the common bile duct
and pancreatic duct that may even lead to pancreatitis.
3. Hypercholesterolemia:
In obstructive jaundice, it is caused by the combination of decreased plasma lecithin cholesterol
acyl transferase (LCAT) activity, leading to leakage of cholesterol and lecithin from ruptured
biliary ductules directly into the bloodstream, and increased hepatic cholesterol synthesis.
Hypercholesterolemia is a major risk factor for cardiovascular diseases.
4. Hypertriglyceridemia:
Refers to a fasting plasma triglyceride measurement that is increased, typically above the 95th
percentile. The situation can fluctuate between hypertriglyceridemia, mild or moderate
hypertriglyceridemia, progressing into severe hypertriglyceridemia. Elevated plasma triglyceride
concentrations contribute to increased risk of cardiovascular disease, both directly with
associated risk factors such as obesity, metabolic syndrome, pro-inflammatory and
prothrombotic biomarkers, and type 2 diabetes mellitus. The increased risk of acute pancreatitis
is an additional consideration when a patient's triglyceride level is very high (typically > 10
mmol/L).

LIPID PROFILE:

Serum lipid profile is measured for cardiovascular risk prediction and has now become almost a
routine test. The test includes four basic parameters: total cholesterol, HDL cholesterol, LDL
cholesterol and triglycerides.
It is usually done in fasting blood specimen. Fasting refers to 12-14 h overnight complete dietary
restriction with the exception of water and medication. Basically fasting state is essential for
triglycerides estimation because it remains high for several hours after meal and the Friedewald
equation is used for calculation of LDL cholesterol

LDL cholesterol = total cholesterol - HDL cholesterol -[triglycerides/5]

It is recommended that healthy adults with no other risk factors for heart disease be tested fasting
lipid profile once every five years. Individuals may also be screened using only a cholesterol test
and not a full lipid profile. For children and adolescents at low risk, lipid testing is usually not
ordered routinely. However, screening with a lipid profile is recommended for children and
youths who are at an increased risk of developing heart disease as adults. Some of the risk factors
are similar to those in adult and include a family history of heart disease or health problems such
as diabetes, high blood pressure (hypertension), or being overweight.

The lipid profile in various age groups is as follows:

LIPID METABOLISM:

Fats taken in the form of food reach the cells adipocytes for storage as triglycerides. The
triacylglycerols upon oxidation yield more than twice as much energy as that of oxidation of
carbohydrates, so they are especially suitable as storage fuels. For use in respiration, a molecule
of triacylglycerol is first hydrolysed by lipase to glycerol and three fatty acid molecules. The
fatty acids and glycerol are released from the cells of adipose tissue into bloodstream. From the
bloodstream, fatty acids are picked up by tissues such as liver, kidneys, lungs, muscles and
adipose tissue for use as fuel and glycerol is taken up by liver, kidneys and intestinal mucosa

FATE OF GLYCEROL
Glycerol, released by lipase action, is phosphorylated by glycerol kinase and the resulting
glycerol 3-phosphate is oxidized to dihydroxyacetone phosphate. The glycolytic enzyme
phosphotriose isomerase converts this compound to glyceraldehyde-3-phosphate which is further
oxidized via glycolysis (see figure 2.16).

Fate of fatty acids:

Fatty acids are oxidized to acetyl~CoA, so as to meet the energy requirement of the body. The
acetyl-CoA, so produced is oxidized to CO2 via citric acid cycle resulting in further energy
conservation. In some species and in some tissues, the acetyl~CoA has alternative fats.
In liver, acetyl~CoA may be converted to ketone bodies which is exported to the brain and other
tissues when glucose is not available.
BETA-OXIDATION:

• Beta-oxidation is the catabolic process by which fatty acid molecules are broken down in the
cytosol in prokaryotes and in the mitochondria in eukaryotes cell lacking mitochondria (e.g.
RBC) to generate acetyl-CoA.
• Acetyl-CoA enters the citric acid cycle while NADH and FADH2, which are co-enzymes, are
used in the electron transport chain. It is referred as “beta oxidation” because the beta carbon of
the fatty acid undergoes oxidation to a carbonyl group.
1. Activation of Fatty acid
2. Transport of fatty acyl coA into mitochondria
3. Beta- oxidation

The fatty acyl chain is shortened by two carbon atoms as a result of these reactions,  FADH2,
NADH, and acetyl Co A are generated. Because oxidation is on the β carbon and the chain is
broken between the α (2)- and β (3)-carbon atoms—hence the name – β oxidation
Fatty acids must first be converted to an active intermediate before they can be catabolized. This
is the only step in the complete degradation of a fatty acid that requires energy from ATP. The
activation of a fatty acid is accomplished in two steps

Activation of fatty acid:

Fatty acid is converted to fatty acyl CoA by thiokinase or fattyacyl CoA synthetase. This reaction
occurs in cytoplasm and activated by ATP and coenzyme A, and fatty acyl-CoA is formed.
Short-chain fatty acids are activated in mitochondria. The ATP is converted to AMP and
pyrophosphate (PPi), which is cleaved by pyrophosphatase to two inorganic phosphates (2 Pi).
Because two high-energy phosphate bonds are cleaved, the equivalent of two molecules of ATP
is used for fatty acid activation.

Transport of fatty acid into mitochondrial matrix:

Fatty acids are activated on the outer mitochondrial membrane, whereas they are oxidized in the
mitochondrial matrix. Activated long-chain fatty acids are transported across the membrane by
conjugating them to carnitine, Carnitine is widely distributed and is particularly abundant in
muscle. Carnitine is obtained from foods, particularly animal-based foods, and via endogenous
synthesis.

Role of carnitine:
1) The acyl group is attached to the hydroxyl group of carnitine to form acyl carnitine. This
reaction is catalyzed by carnitine acyl transferase I
2) Acyl carnitine is then shuttled across the inner mitochondrial membrane by a translocase.
3) The acyl group is transferred back to CoA on the mitochondrial matrix side of the membrane.
This reaction, is catalyzed by carnitine acyl transferase II. Finally, the translocase returns
carnitine to the cytosolic side in exchange for an incoming acyl carnitine. This movement is
through facilitated diffusion.

Steps of beta Oxidation:

Beta-oxidation of saturated FA (palmitic acid):

These steps are repeated until all the carbons of an even-chain fatty acyl-CoA are converted to
acetyl-CoA.
• An oxidation step that produce FADH2
• A hydration step
• A second oxidation step that produce NADH+ H+
• A thiolytic cleavage that release a molecule of acetyl coA.

Step-1 Dehydrogenation:
The first step is the removal of two hydro g e n atoms from the 2(α ) - and 3(β ) - c carbon atoms,
catalyzed by acyl - CoA de hydrogenase and requiring FAD. This results in the formation of
alpha-beta-unsaturated fatty acyl coA or also called as trans-Δ2- enoyl-Co A.

Step-2- Hydration :
Water is added to saturate the double bond and form 3 -hydroxyacyl-CoA, catalyzed by Δ 2-
enoyl-CoA hydratase.

Step-3- dehydrogenation:
The 3-hydroxy derivative undergoes further dehydrogenation on the 3-carbon catalyzed by 3-
hydroxyacylCoA dehydrogenase to form the corresponding 3-ketoacyl-CoA compound. In this
case, NAD+ is the coenzyme involved.
Step-4- Thiolysis :
3-ketoacyl-CoA is split at the 2,3- position by thiolase (3- ketoacyl-CoAthiolase), forming
acetyl-CoA and a new acyl-CoA two carbons shorter than the original acyl-CoA molecule.
The acyl-CoA formed in the cleavage reaction reenters the oxidative pathway at reaction 2. Since
acetylCoA can be oxidized to CO2 and water via the citric acid cycle the complete oxidation of
fatty acids is achieved.

BIOSYNTEHSIS OF FATTY ACID:

The ability to synthesize a variety of lipids is essential to all organisms and major portion of
saturated as well as monounsaturated fatty acids are synthesized within the body of an organism.
In animal cells, palmitic acid is synthesized first and then, undergoes certain modifications to
synthesize other long-chain fatty acids.
Biosynthesis of fatty acids such as palmitic acid requires acetyl~CoA and an input of chemical
energy in the form of ATP and NADPH. Thus, the process of biosynthesis of fatty acids from
acetyl CoA is called De novo synthesis. It occurs in the cytosol of hepatocytes, adipocytes and in
the mammary glands of lactating animals.
As it is discussed, that oxidation of fatty acids takes place in mitochondria, so for the
biosynthesis of fatty acids, acetyl~CoA is to be transported out of mitochondria, i.e. cytosol. But
mitochondrial inner membrane is impermeable to acetyl~CoA, so there is a shuttle for transfer of
acetyl groups from mitochondria to the cytosol.

Transport of Acetyl~CoA to Cytosol:

Intramitochondrial acetyl~CoA first reacts with oxaloacetate to form citrate in the presence of
citrate synthase. Citrate then passes through the inner membrane on the citrate transporter. As the
mitochondrial outer membrane is freely permeable, inside cytosol citrate cleaves in the presence
of citrate lyase to regenerate acetyl~CoA with the help of energy generated by ATP.

Conversion of Acetyl~CoA to Malonyl~CoA:

Biosynthesis requires the participation of three carbon intermediate, malonyl~CoA which is
formed from acetyl~CoA by the action of acetyl~CoA carboxylase.

The Acetyl~CoA carboxylase has three regions:

1. Biotin carrier protein
2. Biotin carboxylase
3. Transcarboxylase
These regions help in the conversion of acetyl~CoA to malonyl~CoA with the consumption of 1
ATP.

Fatty Acid Synthesis:

Fatty acid synthesis requires the presence of fatty acid synthase which is a multienzyme complex
composed of two identical subunits, each one of which contains 6 enzymes and an acyl carrier
protein. Acyl carrier
protein has its SH group for entry of malonyl group during synthesis of fatty acids whereas B-
ketoacyl~ACP synthase has its -SH group for the entry of acetyl group.

1.Fatty acid synthesis starts with transfer of acetyl group of acetyl~CoA to cysteinyl~SH group
of B-ketoacyl~ACP synthase to form Acetyl~S~ACP. This reaction is catalyzed by
acetyl~CoA~ACP transacetylase.

At the same time, malonyl group of malonyl~CoA is transferred to the -SH group of ACP in the
presence of malonyl~CoA~ACP transferase.

2. Formation of fatty acid chain begins with the condensation of acetyl and malonyl groups to form
acetoacetyl~S~ACP. This is catalyzed by the fatty acid synthase.
3. The acetoacetyl~S~ACP formed in the condensation step, now undergoes reduction of carbonyl group
to form ß-hydroxybutyryl~S~ACP. This reaction is catalyzed by fatty acid synthase.

4. ß-hydroxybutyryl-ACP is then dehydrated to yield a, ß-unsaturated acyl~S~ACP in the presence of

fatty acid synthase.

5. o., ß-unsaturated acyl~S~ACP is reduced in the presence of fatty acid synthase to form butyryl~S~ACP.

Butyryl group is now transferred from the -SH group of ACP to cys -SH group of B-ketoacyl-ACP synthas
Cycle again repeats as butyryl group undergoes condensation with malonyl CoA and such seven cycles o
condensation and reduction produces 16 - carbon saturated palmitoyl group bound to ACP. With this
sten palmitate is released from ACP by hydrolytic activity of enzyme. Overall reaction can be written as: