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Nurse’s Biochemistry Important Questions and

Answers

SHORT ANSWERS
Antioxidants?
Antioxidants are molecules that inhibit or quench free radical reactions and
delay or inhibit cellular damage. Though the antioxidant defenses are
different from species to species, the presence of the antioxidant defense is
universal. Antioxidants exists both in enzymatic and non-enzymatic forms in
the intracellular and extracellular environment.
Normal biochemical reactions, increased exposure to the environment, and
higher levels of dietary xenobiotics result in the generation of reactive
oxygen species (ROS) and reactive nitrogen species (RNS). ROS and
RNS are responsible for the oxidative stress in different pathophysiological
conditions.
Cellular constituents of our body are altered in oxidative stress conditions,
resulting in various disease states. The oxidative stress can be effectively
neutralized by enhancing cellular defenses in the form of antioxidants.
Certain compounds act as in vivo antioxidants by raising the levels of
endogenous antioxidant defenses. Expression of genes encoding the
enzymes such as superoxide dismutase (SOD), catalase (CAT),
glutathione peroxidase (GSHPx) increases the level of endogenous
antioxidants.
 Antioxidants can be categorized in multiple ways. Based on their activity,
they can be categorized as enzymatic and non-enzymatic antioxidants.
Enzymatic antioxidants work by breaking down and removing free radicals.
The antioxidant enzymes convert dangerous oxidative products to
hydrogen peroxide (H2O2) and then to water, in a multi-step process in
presence of cofactors such as copper, zinc, manganese, and iron. Non-
enzymatic antioxidants work by interrupting free radical chain reactions.
Few examples of the non-enzymatic antioxidants are vitamin C, vitamin E,
plant polyphenol, carotenoids, and glutathione.

The other way of categorizing the antioxidants is based on their solubility in

the water or lipids. The antioxidants can be categorized as water-soluble
and lipid-soluble antioxidants. The water-soluble antioxidants (e.g. vitamin
C) are present in the cellular fluids such as cytosol, or cytoplasmic matrix.
The lipid-soluble antioxidants (e.g. vitamin E, carotenoids, and lipoic acid)
are predominantly located in cell membranes.
The antioxidants can also be categorized according to their size, the small-
molecule antioxidants and large-molecule antioxidants. The small-molecule
antioxidants neutralize the ROS in a process called radical scavenging and
carry them away. The main antioxidants in this category are vitamin C,
vitamin E, carotenoids, and glutathione (GSH). The large-molecule
antioxidants are enzymes (SOD, CAT, and GSHPx) and sacrificial proteins
(albumin) that absorb ROS and prevent them from attacking other essential
proteins.

Biochemical functions of calcium?

1. Bone mineralization
2. Blood coagulation
3. Bone and teeth formation
 4. Muscle contraction
Calcium promotes blood coagulation. When cell injured, calcium ions
stimulate the release of certain factor that activates the enzyme
thrombokinase. Thrombokinase catalyzes the conversion of Prothrombin to
thrombin, which needed in blood clotting.
 Sending and receiving nerve signals
 Releasing hormones and other chemicals
 Keeping a normal heartbeat
In the teeth, the mineral occurs also in the form of hydroxyapatite in the
enamel and dentine but crystals formed are much larger which accounts for
the difficulty of withdrawing it during Deprivation.
When the calcium ions activate the myosin and actin, energy is released
from ATP which causes Contraction.
In the long run of the growth process, the bone shaft which supports the
weight of the bone is lengthened with the formation of new matrix.
The calcium ions are again bound back on the reticulum causing
Relaxation.

Competitive Inhibition of enzymes?

In competitive inhibition, the substrate and inhibitor cannot bind to the
enzyme at the same time. This usually results from the inhibitor having an
affinity for the active site of an enzyme where the substrate also binds; the
substrate and inhibitor compete for access to the enzyme's active site. This
type of inhibition can be overcome by sufficiently high concentrations of
substrate (Vmax remains constant), i.e., by out-competing the inhibitor.
However, the apparent Km will increase as it takes a higher concentration of
the substrate to reach the Km point, or half the Vmax. Competitive inhibitors
are often similar in structure to the real substrate
 Functions of Ascorbic acid?
Vitamin C, also known as ascorbic acid and L-ascorbic acid, is
a vitaminfound in various foods and sold as a dietary supplement. It is used
to prevent and treat scurvy. Vitamin C is an essential nutrient involved in
the repair of tissue and the enzymatic production of
certain neurotransmitters. It is required for the functioning of several
enzymes and is important for immune system function. It also functions as
an antioxidant.

Normal range - Random blood glucose, serum cholesterol, serum

bilirubin?

Random Blood glucose

The reference values for a "normal" random glucose test in an average

adult are 79–160mg/dl (4.4–8.9 mmol/l), between 160-200mg/dl (8.9–
11.1 mmol/l) is considered pre-diabetes, and > 200 mg/dl is considered
diabetes according to ADA guidelines.
Serum Cholesterol

 Desirable: Less than 200 mg/dL

 Borderline high: 200-239 mg/dL

 High: Over 240 mg/dL

Having a total cholesterol level over 240 mg/dL may double the risk of heart
disease.

Serum Bilirubin
A normal level is:

 Direct (also called conjugated) bilirubin: less than 0.3 mg/dL (less
than 5.1 µmol/L)
 Total bilirubin: 0.1 to 1.2 mg/dL (1.71 to 20.5 µmol/L)
Deamination?
Deamination is the removal of an amine group from
a molecule. Enzymes that catalyse this reaction are called deaminases.
In the human body, deamination takes place primarily in the liver
however glutamate is also deaminated in the kidneys. In situations of
excess protein intake, deamination is used to break down amino acids for
energy. The amine group is removed from the amino acid and converted
to ammonia. The rest of the amino acid is made up of
mostly carbon and hydrogen, and is recycled or oxidized for energy.
Ammonia is toxic to the human system, and enzymes convert it
to urea or uric acid by addition of carbon dioxide molecules (which is not
considered a deamination process) in the urea cycle, which also takes
place in the liver. Urea and uric acid can safely diffuse into the blood and
then be excreted in urine.

Essential Fatty Acids?

Essential fatty acids, or EFAs, are fatty acids that humans and other
animals must ingest because the body requires them for good health but
cannot synthesize them.
The term "essential fatty acid" refers to fatty acids required for biological
processes but does not include the fats that only act as fuel.
Essential fatty acid: An unsaturated fatty acid that is essential to human
health, but cannot be manufactured in the body. There are three types of
EFAs: arachnoidic acid, linoleic acid, and linolenic acid. When linoleic acid
is obtained in the diet, it can be converted to both arachnoidic and linolenic
acid. Linoleic acid is commonly found in cold-pressed oils, especially oils
extracted from cold-water fish and certain seeds. Supplementation with
EFAs appears to be useful as a treatment for certain neurological
disorders.

Function of Thiamine?
Thiamin is one of the B vitamins. The B vitamins are a group of water-
soluble vitamins that are part of many of the chemical reactions in the body.
Function
Thiamin (vitamin B1) helps the body's cells change carbohydrates into
energy. The main role of carbohydrates is to provide energy for the body,
especially the brain and nervous system.

Thiamin also plays a role in muscle contraction and conduction of nerve

signals.

Thiamin is essential for the metabolism of pyruvate.

Three enzymes of diagnostic importance?

1. Aspartate Transminase (AST)

2. Alanine Transaminase (ALT)

3. Gamma Glutamyl Transferase (GGT)

4. Alkaline Phosphatase (ALP)

5. Pancreatic amylase and lipase

6. Creatine Kinase (CK)

Enzymes: Chemical substances which act as biological catalysts and

augment the rate of specific metabolic and biochemical reaction within the
body.

Clinical Importance of Enzymes

Enzymes: Diagnostic markers of diseases. Principle:

Concentration/Localization of specific enzymes in blood may vary in
response to abnormal/diseased conditions.

Clinical Diagnosis of a disease using Enzyme Markers:

 1. Presence/absence of the marker enzymes.

2. Change in enzyme concentration – plasma/serum/cellular

concentration.

3. Change in enzyme localization – leakage from cell to blood.

4. Pattern of enzyme clearance – abnormal accumulation.

Normal serum level - sodium, potassium and calcium?

Serum Calcium - The total serum calcium concentration is normally 8.5-

10.2 mg/dL,

Serum Sodium - A normal blood sodium level is between 135 and 145
milliequivalents per liter (mEq/L).

Serum Potassium - The normal potassium level in the blood is 3.5-5.0

milliEquivalents per liter (mEq/L). Potassium levels between 5.1 mEq/L to
6.0 mEq/L are considered to be mild hyperkalemia.

Starch?

Starch or amylum is a polymeric carbohydrate consisting of a large

number of glucose units joined by glycosidic bonds. This polysaccharide is
produced by most green plants as energy storage. It is the most common
carbohydrate in human diets and is contained in large amounts in staple
foods like potatoes, wheat, maize (corn), rice, and cassava.
Pure starch is a white, tasteless and odorless powder that is insoluble in
cold water or alcohol. It consists of two types of molecules: the linear
and helical amylose and the branched amylopectin. Depending on the
plant, starch generally contains 20 to 25% amylose and 75 to 80%
amylopectin by weight. Glycogen, the glucose store of animals, is a more
highly branched version of amylopectin.
Heat Coagulation Test?

Uses: Screening test for proteinuria in pregnancy.

When a protein is heated,its physical,chemical and biological properties are

changed due to breaking up of certain bonds.the conformation of protein
will be changed. This process is known as denaturation.

Procedure:

 Fill two third of test tube with the given solution.

 Add 1 to 2 drops of chlorophenol red indicator drop by drop and mix.

 When purple red colour develops,add 1 % acetic acid drop by drop

untill the colour changes to faint pink.

 Hold the test tube near its bottom,incline it slightly and heat the upper
portion of the fluid.

Application:

This is the most widely used test for the detection of proteins in the urine.

Hyperglycemia?

Hyperglycemia – elevated blood glucose levels.

Hyperglycemia is a complex metabolic condition characterized by

abnormally high levels of blood sugar (blood glucose) in circulating blood,
usually as a result of diabetes mellitus (types 1 and 2).

Hyperglycemia, also known as diabetic ketoacidosis, is a condition that

develops over a period of a few days as the blood glucose levels of a type
1 or type 2 diabetic gradually rise. Ketoacidosis occurs when increasing
glucose levels are met by a lack of sufficient or effective insulin production,
starting a sequence of physiologic events as follows:
  The combination of excess glucose production and low glucose
utilization in the body raises levels of blood glucose, which leads to
increased urinary output (diuresis) followed quickly by a loss of fluid
and essential mineral salts (electrolytes) and, ultimately, dehydration.
The loss of fluid may finally result in dehydration. If the entire process
is severe enough over several hours (serum glucose levels over
800mg/dL), swelling can occur in the brain (cerebral edema), and
coma can eventually result.

 In a metabolic shift to a catabolic (breaking down) process, cells

throughout the body empty their electrolytes (sodium, potassium, and
phosphate) into the bloodstream. Electrolytes control the fluid
balance of the body and are important in muscle contraction, energy
generation, and almost all major biochemical reactions in the body.
As a result of electrolyte imbalance, many functions can become
impaired.

 Free fatty acids from lipid stores are increased, encouraging the
production of ketoacids in the liver, leading to an over-acidic condition
(metabolic acidosis) that causes even more disruption in body
processes.

Without effective treatment of the hyperglycemic episode, the child can

lapse into a diabetic coma, which sometimes leads to death.

Essential Aminoacids?

Protein is a essential agent of biological function.There are 22

standard amino acids, but only 21 are found in eukaryotes. Of the 22, 20
are directly encoded by the universal gentic code. Humans can synthesize
11 of these 20 from each other or from other molecules of intermediary
metabolism. The other 9 must be consumed in the diet, and so are called
essential amino acids.

Essential amino acids, which generally have a longer half-life than the
nonessential ones, are those that are required in the diet since the body
cannot synthesize them in adequate amounts to
maintain protein biosynthesis. If even one essential (or nones-
sential) amino acid is absent, the remaining 19 cannot be used, and they
become catabolized thus leading to a negative nitrogen balance. Essential
amino acids vary depending on species and age.

Essential aminoacids are histidine, isoleucine, leucine, lysine, methionine,

phenylalanine, threonine, tryptophan, and valine. The remaining
two, selenocysteine and pyrrolysine, are incorporated into proteins by
unique synthetic mechanisms.

Bile Salts and its Function?

Bile salts help in solubilize and digest lipids.

As amphipathic molecule with hydrophobic and hydrophilic regions,

conjugated bile salts sit at the lipid/water interface and, at the right
concentration, form micelles. The added solubility of conjugated bile salts
aids in their function by preventing passive re-absorption in the small
intestine.

As a result, the concentration of bile acids/salts in the small intestine is high

enough to form micelles and solubilize lipids. "Critical micellar
concentration" refers to both an intrinsic property of the bile acid itself and
amount of bile acid necessary to function in the spontaneous and dynamic
formation of micelles. Bile acid-containing micelles aid lipases to digest
lipids and bring them near the intestinal brush border membrane, which
results in fat absorption.

Bile acids have other functions, including eliminating cholesterol from the
body, driving the flow of bile to eliminate certain catabolites
(including bilirubin), emulsifying fat-soluble vitamins to enable their
absorption, and aiding in motility and the reduction of the bacteria flora
found in the small intestine and biliary tract.
Bile acids have metabolic actions in the body resembling those
of hormones, acting through two specific receptors, the farnesoid X
receptor and G protein-coupled bile acid receptor/TGR5. They bind less
specifically to some other receptors and have been reported to regulate the
activity of certain enzymes and ion channels and the synthesis of diverse
substances including endogenous fatty acid ethanolamides.
Hypercholesterolemia?

Hypercholesterolemia, also called high cholesterol, is the presence of high

levels of cholesterol in the blood. It is a form of hyperlipidemia, high blood
lipids, and hyperlipoproteinemia (elevated levels of lipoproteins in the
blood).
Elevated levels of non-HDL cholesterol and LDL in the blood may be a
consequence of diet, obesity, inherited (genetic) diseases (such as LDL
receptor mutations in familial hypercholesterolemia), or the presence of
other diseases such as type 2 diabetes and an underactive thyroid.

Hypercholesterolemia can be defined as the presence of

high plasma cholesterol levels, with normal plasma triglycerides, as a
consequence of the rise of cholesterol and apolipoprotein B (apoB)-
rich lipoproteins, called low-density lipoprotein (LDL).

Mitochondria?

The purpose of the mitochondria in the eukaryote is to provide cellular

respiration to the cell.
Mitochondria are commonly between 0.75 and 3 μm in diameter but vary
considerably in size and structure. Unless specifically stained, they are not
visible. In addition to supplying cellular energy, mitochondria are involved in
other tasks, such as signaling, cellular differentiation, and cell death, as
well as maintaining control of the cell cycle and cell growth. The number of
mitochondria in a cell can vary widely by organism, tissue, and cell type.
For instance, red blood cells have no mitochondria, whereas liver cells can
have more than 2000. The organelle is composed of compartments that
carry out specialized functions. These compartments or regions include the
outer membrane, the intermembrane space, the inner membrane, and
the cristae and matrix.
A mitochondrion contains outer and inner membranes composed
of phospholipid bilayers and proteins. The two membranes have different
properties. Because of this double-membraned organization, there are five
distinct parts to a mitochondrion. They are:

1. the outer mitochondrial membrane,

2. the intermembrane space (the space between the outer and inner
membranes),

3. the inner mitochondrial membrane,

4. the cristae space (formed by infoldings of the inner membrane), and

5. the matrix (space within the inner membrane).

Rothera's test?

A test for ketone bodies; 5 mL of fresh urine are saturated with solid ammo
nium sulfate andmixed with 10 drops of freshly prepared 2% sodium nitropr
usside solution, which is thenmixed with 10 drops of concentrated ammonia
water and allowed to stand for 15 minutes; the
presence of acetoacetic acid, or of larger concentrations of acetone, is indic
ated by thedevelopment of a blue-purple color.

Principle:
Acetoacetic acid and acetone react with alkaline solution of sodium
nitroprusside to form a purple colored complex. This method can detect
above 1-5 mg/dl of acetoacetic acid and 10-20 mg/dl of acetone. Beta-
hydroxybutyrate is not detected.

Requirements:

1. Urine specimen
2. Test tubes
3. Rothera’s powder - Sodium nitroprusside = 0.75 gm,
Ammonium sulphate = 20g. Mix and pulverize.
Liquor ammonia (Ammonium hydroxide

Procedure:
1. Transfer about 5 ml of urine to a test tube.
2. Add 1 gm of Rothera’s powder mixture and mix well.
3. Layer over the urine 1-2 ml of concentrated ammonium hydroxide.
4. Observe the pink-purple ring at the interface.

Observation and Results:

 Immediate formation of purple permanganate colored ring at the
interface: Ketone bodies present (Positive)
 No formation of purple permanganate colored ring at the interface:
ketone bodies absent (Negative)
Grade the result according to intensity of the formation of colored ring as
Trace, +, ++, +++ or ++++.

Ketone Bodies?

Ketone bodies are the water-

soluble molecules (acetoacetate, beta-hydroxybutyrate, and the
spontaneous breakdown product of acetoacetate, acetone)
containing the ketone group that are produced by the
liver from fatty acids during periods of low food intake
(fasting), carbohydrate restrictive diets, starvation, prolonged
intense exercise, alcoholism or in untreated (or inadequately
treated) type 1 diabetes mellitus.

Ketone bodies, or simply ketones are substances produced by

the liver during gluconeogenesis, a process which creates glucose
in times of fasting and starvation. There are three ketone bodies
produced by the liver. They are acetoacetate, beta-
hydroxybutyrate, and acetone. These compounds are used in
healthy individuals to provide energy to the cells of the body when
glucose is low or absent in the diet.

Metabolic Acidosis?

Metabolic acidosis is primary reduction in bicarbonate, typically with

compensatory reduction in carbon dioxide partial pressure; pH may be
markedly low or slightly subnormal.

Metabolic acidoses are categorized as high or normal anion gap based on

the presence or absence of unmeasured anions in serum. Causes include
accumulation of ketones and lactic acid, renal failure, and drug or toxin
ingestion (high anion gap) and GI or renal bicarbonate loss. Symptoms and
signs in severe cases include nausea and vomiting, lethargy and
hyperpnea.
HMP Shunt Pathway?

Glucose can alternatively also undergo a different pathway to produce

other products required by the cells. One of these alternate pathway is
the pentose phosphate pathway or also called as hexose
monophosphate pathway in which oxidation of glucose 6-phosphate
takes place to produce pentoses. The fate of glucose whether to undergo
glycolysis or the hexose monophosphate pathway is decided by the relative
concentrations of NADP+ and NADPH. Hexose monophosphate shunt is
useful in adipose tissue, liver, erythrocytes, testes, adrenal glands and
lactating mammary glands.

This metabolic pathway is fragmented into two phases taking place in the
cytosol as all the enzymes required are present there: the oxidative and
the non-oxidative phase.

The pentose phosphate pathway takes place in the cytosol of the cell, the
same location as glycolysis. The two most important products from this
process are the ribose-5-phosphate sugar used to make DNA and RNA,
and the NADPH molecules which help with building other molecules.

Immunoglobulin G?

IgG is the most common class of immunoglobulin. It is present in the

largest amounts in blood and tissue fluids. Each IgG molecule consists of
the basic four-chain immunoglobulin structure—two identical H chains and
two identical L chains (either kappa or lambda)—and thus carries two
identical antigen-binding sites. There are four subclasses of IgG, each with
minor differences in its H chains but with distinct biological properties. IgG
is the only class of immunoglobulin capable of crossing the placenta;
consequently, it provides some degree of immune protection to the
developing fetus. These molecules also are secreted into the mother’s milk
and, once they have been ingested by an infant, can be transported into
the blood, where they confer immunity.

Pellagra?

Pellagra is a nutritional disease due to deficiency of the vitamin niacin and

the essential amino acidtryptophan. The clinical
features of pellagra are dermatitis, diarrhea, and dementia; it is commonly
known as the ‘disease of the four Ds,’ since it is also fatal – the fourth ‘D’
is death.

Beri Beri?
Thiamine deficiency, or beriberi, refers to the lack of thiamine
pyrophosphate, the active form of the vitamin known as thiamine (also
spelled thiamin), or vitamin B-1. Thiamine pyrophosphate, the biologically
active form of thiamine, acts as a coenzyme in carbohydrate metabolism
through the decarboxylation of alpha ketoacids. It also takes part in the
formation of glucose by acting as a coenzyme for the transketolase in the
pentose monophosphate pathway.
Benedict’s Test?
Benedict’s Test is used to test for simple carbohydrates. The Benedict’s
test identifies reducing sugars (monosaccharide’s and some
disaccharides), which have free ketone or aldehyde functional
groups. Benedict’s solution can be used to test for the presence of glucose
in urine.
Some sugars such as glucose are called reducing sugars because they are
capable of transferring hydrogens (electrons) to other compounds, a
process called reduction. When reducing sugars are mixed with Benedicts
reagent and heated, a reduction reaction causes the Benedicts reagent to
change color. The color varies from green to dark red (brick) or rusty-
brown, depending on the amount of and type of sugar.
Normal level of blood urea and fasting blood sugar?

The normal ranges for these biomarkers were 70 – 110 mg/dl and 110 - 140
mg/dl for fasting and postprandial blood sugar respectively; 15-40mg/dl
for serum urea; and 0.6 - 1.2 mg/dl and 0.5 - 1.1 mg/dl for serum
creatinine for males /females respectively.

Transamination?
Transamination, a chemical reaction that transfers an amino group to
a ketoacid to form new amino acids. This pathway is responsible for the
deamination of most amino acids. This is one of the major degradation
pathways which convert essential amino acids to nonessential amino
acids (amino acids that can be synthesized de novo by the organism).
Transamination in biochemistry is accomplished by enzymes called
transaminases or aminotransferases. α-ketoglutarate acts as the
predominant amino-group acceptor and produces glutamate as the new
amino acid.
Aminoacid + α-ketoglutarate ↔ α-keto acid + Glutamate
Glutamate's amino group, in turn, is transferred to oxaloacetate in a
second transamination reaction yielding aspartate.
Glutamate + oxaloacetate ↔ α-ketoglutarate + aspartate

Metabolic Alkalosis?
Metabolic alkalosis is a primary pathophysiologic event characterized by
the gain of bicarbonate or the loss of nonvolatile acid from extracellular
fluid such that arterial pH increases (normal pH = 7.40). More simply put,
it is a primary increase in plasma bicarbonate concentration (normal
plasma HCO3 = 24mEq/L).

Iodine?
In vertebrate biology, iodine's primary function is as a constituent of the
thyroid hormones, thyroxine (T4) and triiodothyronine (T3). These
molecules are made from addition-condensation products of the amino acid
tyrosine, and are stored prior to release in an iodine-containing protein
called thyroglobulin.

Normal level - FBS, blood urea, Serum total protein and SGOT?

A fasting blood sugar level less than 100 mg/dL (5.6 mmol/L) is normal. A
fasting blood sugar level from 100 to 125 mg/dL (5.6 to 6.9 mmol/L) is
considered prediabetes. If it's 126 mg/dL (7 mmol/L) or higher on two
separate tests, you have diabetes.
Blood Urea - In general, around 7 to 20 mg/dL (2.5 to 7.1 mmol/L) is
considered normal.
Serum Total Protein - The normal serum protein level is 6 to 8 g/dl.
Albumin makes up 3.5 to 5.0 g/dl, and the remainder is the total globulins.
SGOT - The normal range of values for AST (SGOT) is about 5 to 40 units
per liter of serum (the liquid part of the blood). The normal
range of values for ALT (SGPT) is about 7 to 56 units per liter of serum.

Proteolytic enzymes examples?

Proteolytic enzyme, also called protease, proteinase, or peptidase, any of
a group of enzymes that break the long chainlike molecules of proteins into
shorter fragments (peptides) and eventually into their components, amino
acids.

Coenzyme form of thiamine and pyridoxine?

Thiamine - thiamine pyrophosphate.
Pyridoxine - pyridoxal phosphate
vitamin B 2 (riboflavin) - flavin mononucleotide or flavin adenine
dinucleotide
vitamin B 3 (niacin) - nicotinamide adenine dinucleotide or nicotinamide
adenine dinucleotide phosphate.

Lactose Intolerance?
Lactose intolerance is a digestive disorder caused by the inability to
digest lactose, the main carbohydrate in dairy products. It can cause
various symptoms, including bloating, diarrhea and abdominal cramps.
People with lactose intolerance don't make enough of the enzyme
lactase, which is needed to digest lactose.
Dietary Fibre?
Dietary fibre is a type of carbohydrate that cannot be digested by our
bodies' enzymes. It is found in edible plant foods such as cereals, fruits,
vegetables, dried peas, nuts, lentils and grains. Fibre is grouped by its
physical properties and is called soluble, insoluble or resistant starch.

Daily requirement of vit A and D?

Vitamin D intake is recommended at 400–800 IU/day, or 10–20

micrograms. However, some studies suggest that a higher daily intake of
1000–4000 IU (25–100 micrograms) is needed to maintain optimal blood
levels.
The Recommended Dietary Allowance (RDA) for men and women is 900
and 700 μg retinol activity equivalents (RAE)/day, respectively. The
Tolerable Upper IntakeLevel (UL) for adults is set at 3,000 μg/day of
preformed vitamin A.

Primary structure of proteins?

The primary structure of a protein refers to the sequence of amino acids

in the polypeptide chain. The primary structure is held together by peptide
bonds that are made during the process of protein biosynthesis.
Active Transport?
The movement of ions or molecules across a cell membrane into a region
of higher concentration, assisted by enzymes and requiring energy.

Glycosuria?
Glycosuria is the excretion of glucose into the urine. Ordinarily, urine
contains no glucose because the kidneys are able to reabsorb all of the
filtered glucose from the tubular fluid back into the bloodstream.

Rickets?
Rickets is the softening and weakening of bones in children, usually
because of an extreme and prolonged vitamin D deficiency. Rare inherited
problems also can cause rickets.

Because rickets softens the areas of growing tissue at the ends of a child's
bones (growth plates), it can cause skeletal deformities such as:

 Bowed legs or knock knees

 Thickened wrists and ankles

 Breastbone projection

Provitamin Example?
Vitamin A and Vitamin B6
A provitamin is a substance that may be converted within the body to
a vitamin. "Provitamin B5" is a name for panthenol, which may be
converted in the body to vitamin B5 (pantothenic acid).
"provitamin A" is a name for β-carotene, which has only about 1/6 the
biological activity of retinol (vitamin A); the body uses an enzyme to convert
β-carotene to retinol.

Marker Enzyme in Myocardial infarction?

Cardiac enzymes ― also known as cardiac biomarkers ― include
myoglobin, troponin and creatine kinase. Historically, lactate
dehydrogenase, or LDH, was also used but is non-specific.

Golgi Apparatus?
The Golgi apparatus, also known as the Golgi complex, Golgi body, or
simply the Golgi, is an organelle found in most eukaryotic cells. It was
identified in 1897 by the Italian scientist Camillo Golgi and named after him
in 1898.
Part of the endomembrane system in the cytoplasm, the Golgi
apparatus packages proteins into membrane-bound vesicles inside the cell
before the vesicles are sent to their destination. The Golgi apparatus
resides at the intersection of the secretory, lysosomal, and endocytic
pathways. It is of particular importance in processing proteins for secretion,
containing a set of glycosylation enzymes that attach various sugar
monomers to proteins as the proteins move through the apparatus.

Electrophoresis?
Electrophoresis is a separation technique that is normally performed in an
aqueous environment. This is due to the fact that the separation
mechanism is based on the difference in migration rate of
charged species in an electric field. Species (ions/molecules or particles)
with a difference in their charge over size ratio will exhibit a difference in
migration rate. Most charged species are fairly soluble in aqueous media
and thus water is the most obvious solvent for electrophoresis. However, in
a number of non aqueous solvent systems, it is possible to obtain
sufficient conductivity to perform electrophoresis. If such systems are
utilized with the technique of capillary electrophoresis, a number of
advantages compared to aqueous systems are obtained in the separation
of small molecules.

Sodium and Potassium Pump?

The process of moving sodium and potassium ions across the cell
membrane is an active transport process involving the hydrolysis of ATP to
provide the necessary energy. It involves an enzyme referred to as Na +/K+-
ATPase. This process is responsible for maintaining the large excess of
Na+ outside the cell and the large excess of K+ ions on the inside. A cycle of
the transport process is sketched below. It accomplishes the transport of
three Na+ to the outside of the cell and the transport of two K + ions to the
inside. This unbalanced charge transfer contributes to the separation of
charge across the membrane. The sodium-potassium pump is an important
contributer to action potential produced by nerve cells. This pump is called
a P-type ion pump because the ATP interactions phosphorylates the
transport protein and causes a change in its conformation

Brief Answers (5 Marks)

Urea Cycle?

THE UREA CYCLE

SYNTHESIS OF CARBAMOYL PHOSPHATE: This is how we "activate" a

free ammonia before we subsequently make urea.
  Enzyme = Carbamoyl Phosphate Synthetase.

 This reaction occurs in the mitochondrion.

 2 ATP are required. Basically these are used to "charge" or "activate"

ammonia with a high-energy phosphate bond, before we
subsequently start urea synthesis.

 N-Acetylglutamate is absolutely required as a cofactor. This

compound also serves a regulatory role in urea synthesis.

o The rate of carbamoyl phosphate synthesis is dependent on the

levels of N-Acetylglutamate in the mitochondria (I'm not sure
whether this is a linear relationship).

 Usually, the free ammonia is derived directly from Glutamate

Dehydrogenase, but it could come from anywhere.

THE UREA CYCLE:

 REQUIRED STARTING MATERIALS:

o Carbamoyl Phosphate: It donates a free NH3 to urea, per turn of

the cycle.

o Ornithine: It is regenerated each turn of the cycle

o Aspartic Acid: It donates an NH3 to urea from

aminotransferases, per turn of the cycle.

 Aspartic Acid has the same carbon skeleton as

Oxaloacetate:
 o 4 ATP are required per molecule of urea

 2ATP required to synthesize Carbamoyl Phosphate

 2ATP required to synthesize Arginosuccinate, via 1 ATP

and a pyrophosphatase

 Carbamoyl Phosphate + Ornithine ------> Citrulline

o This is THE COMMITTED STEP

o One of the two NH3's of urea comes to us from Carbamoyl

Phosphate, essentially the same as starting with a charged free
NH3.

o This step occurs in the mitochondria. Citrulline is then

transferred from mitochondria to the cytosol for the rest of the
steps.

o Enzyme: Ornithine Transcarbamoylase

 Citrulline + Aspartic Acid ------> Arginosuccinate

o ATP ------> AMP + PPi. Metabolic equivalent of 2ATP required,

with pyrophosphatase driving the reaction forward, making the
reaction irreversible.

o Aspartic Acid carries with it the other NH3 of urea. This

NH3 has come to us via Aspartic Acid, courtesy of Aspartate
Aminotransferase.

o This step, and all subsequent steps, occurs in the cytosol.

 Arginosuccinate ------> Arginine + Fumarate

o Catalyzed by Arginosuccinase
 o Fumarate then goes through the TCA Cycle, in order
to regenerate the carbon skeleton of Aspartic Acid:

 Fumarase: Fumarate ------> L-Malate

 L-Malate Dehydrogenase: L-Malate ------> Oxaloacetate

 Aspartate Aminotransferase: Oxaloacetate + NH3 ------

> Aspartic Acid

 Arginine ------> Urea + Ornithine

o Catalyzed by Arginase. Know this enzyme, because this is the

step that regenerates ornithine.

 END PRODUCTS:

o Fumarate, which is recycled, via the TCA cycle and

transamination, to Aspartate

o Urea

o Ornithine, which is recycled to the first step.

REGULATION OF THE UREA CYCLE:

 Short-Term Regulation (metabolic regulation):

o The primary regulated step is Carbamoyl Phosphate

Synthetase.

o Increased levels of Arginine promotes the formation of N-

Acetyl Glutamate, which is required for carbamoyl phosphate
synthesis.

 Glutamate + Acetyl-CoA ------> N-Acetylglutamate

o So when Arginine levels goes up, carbamoyl phosphate is

made and urea synthesis occurs.
  Long-Term Regulation (diet)

o In the long-term, a high-protein diet will increase overall levels

of urea synthesis.

o People who are protein derived would be ill-equipped to handle

a sudden massive increase in the amount of protein in their
diet, because enzyme levels would not be high enough. That's
why you change a diet gradually as a rule of thumb, especially
for the malnourished.

AMMONIA DETOXIFICATION: Ways of getting rid of excess ammonia in

the bloodstream.

 From Benzoic Acid: HIPPURATE

o Benzoic Acid + Coenzyme-A ------> Benzoyl-CoA

 2 ATP are required (coupled with pyrophosphatase), as

usual, to make a CoA derivative.

o Benzoyl-CoA + Glycine ------> Hippurate

 Amino groups are fed in at this step, in the form of

Glycine.

o Hippurate is innocuous and is readily excreted in the urine.

o This form of detoxification will only work if we can transaminate

an NH3 group onto glyoxalase (the keto-acid form of glycine). If
we can't get the NH3 into glycine, then we can't get rid of it by
this means!

 From Phenylacetate: PHENYLACETYLGLUTAMINE

o Phenylacetate + Glutamine ------> Phenylacetylglutamine

o Coenzyme-A is required as a cofactor, but there is no free

Coenzyme-A intermediate. In other words, it is a concerted
reaction.
 o 2 ATP are required (coupled with pyrophosphatase), as usual.

o Phenylacetylglutamine is readily excreted in the urine.

CLINICAL CASE-STUDY: HYPERAMMONEMIA

 The patient had a deficiency in Ornithine Transcarbamoylase.

o Blood-levels of NH3 were almost twice normal.

o The enzyme was not completely absent, however. If you gave

the patient protein, it would take him 120 hours to completely
metabolize the protein to urea, which is far longer than normal.

 CLINICAL PRESENTATION: Bizarre behavior: crying, agitation,

babbling, loss of sense of reality.

 Lab results suggest a deficiency in ornithine transcarbamoylase:

o High levels of Alanine, Glutamine, and Orotic Acid (a metabolite

of purine synthesis)

o NH3 of course was way high

o The Carbamoyl Phosphate builds up and must be shunted off to

another pathway.

 TREATMENT:

o A high carbohydrate, low protein diet (but not no protein)

o Treat with benzoic acid or phenylacetate for ammonia

detoxification.

Formation and Utilization of Ketone Bodies?

Formation of Ketone Bodies
The acetyl-CoA produced by β-oxidation enters the citric acid cycle in the
mitochondrion by combining with oxaloacetate to form citrate. This results
in the complete combustion of the acetyl group of acetyl-CoA to CO 2 and
water. The energy released in this process is captured in the form of
1 GTP and 11 ATP molecules per acetyl group (or acetic acid molecule)
oxidized. This is the fate of acetyl-CoA wherever β-oxidation of fatty acids
occurs, except under certain circumstances in the liver. In the liver
oxaloacetate is wholly or partially diverted into the gluconeogenic
pathway during fasting, starvation, a low carbohydrate diet, prolonged
strenuous exercise, and in uncontrolled type 1 diabetes mellitus.
Under these circumstances oxaloacetate is hydrogenated to malate which
is then removed from the mitochondrion to be converted into glucose in the
cytoplasm of the liver cells, from where the glucose is released into the
blood. In the liver, therefore, oxaloacetate is unavailable for condensation
with acetyl-CoA when significant gluconeogenesis has been stimulated by
low (or absent) insulin and high glucagon concentrations in the blood.
Under these circumstances acetyl-CoA is diverted to the formation
of acetoacetate and beta-hydroxybutyrate. Acetoacetate, beta-
hydroxybutyrate, and their spontaneous breakdown product, acetone, are
known as ketone bodies.

The ketone bodies are released by the liver into the blood. All cells with
mitochondria can take ketone bodies up from the blood and reconvert them
into acetyl-CoA, which can then be used as fuel in their citric acid cycles,
as no other tissue can divert its oxaloacetate into the gluconeogenic
pathway in the way that the liver does this. Unlike free fatty acids, ketone
bodies can cross the blood-brain barrier and are therefore available as fuel
for the cells of the central nervous system, acting as a substitute for
glucose, on which these cells normally survive.The occurrence of high
levels of ketone bodies in the blood during starvation, a low carbohydrate
diet and prolonged heavy exercise can lead to ketosis, and in its extreme
form in out-of-control type 1 diabetes mellitus, as ketoacidosis.
Acetoacetate has a highly characteristic smell, for the people who can
detect this smell, which occurs in the breath and urine during ketosis. On
the other hand, most people can smell acetone, whose "sweet & fruity"
odor also characterizes the breath of persons in ketosis or,
especially, ketoacidosis.
Utilization of Ketone Bodies
Ketone bodies can be utilized as fuel in the heart, brain and muscle, but not
the liver. They yield 2 guanosine triphosphate(GTP) and 22 adenosine
triphosphate (ATP) molecules per acetoacetate molecule when oxidized in
the mitochondria. Ketone bodies are transported from the liver to other
tissues, where acetoacetate and beta-hydroxybutyrate can be reconverted
to acetyl-CoA to produce reducing equivalents (NADH and FADH 2), via
the citric acid cycle.
Though it is the source of ketone bodies, the liver itself cannot use them for
energy because it lacks the enzyme thiophorase (β-ketoacyl-CoA
transferase). Acetone is taken up by the liver in low concentrations and
undergoes detoxification through the methylglyoxal pathway which ends
with lactate.
Acetone in high concentrations, as can occur with prolonged fasting or a
ketogenic diet, is absorbed by cells outside the liver and metabolized
through a different pathway via 1,2-propanediol. Though the pathway
follows a different series of steps requiring ATP, 1,2-propanediol can
eventually be turned into pyruvate.
The heart preferentially utilizes fatty acids as fuel under normal physiologic
conditions. However, under ketotic conditions, the heart can effectively
utilize ketone bodies for this purpose.
The brain gets a portion of its fuel requirements from ketone bodies
when glucose is less available than normal (e.g., during fasting,
strenuous exercise, low carbohydrate, ketogenic diet and in neonates). In
the event of a low glucose concentration in the blood, most other tissues
have alternative fuel sources besides ketone bodies and glucose (such as
fatty acids), but current research indicates that the brain has an obligatory
requirement for some glucose. After the diet has been changed to lower
blood glucose utilization for 3 days, the brain gets 25% of its energy from
ketone bodies.
After about 24 days, ketone bodies become the major fuel of the brain,
making up to two-thirds of brain fuel consumption. Many studies suggest
that human brain cells can survive with little or no glucose, but proving the
point is ethically questionable. During the initial stages the brain does not
burn ketones, since they are an important substrate for lipid synthesis in
the brain. Furthermore, ketones produced from omega-3 fatty acids may
reduce cognitive deterioration in old age.
 Renal regulation of acid base balance?
The organs involved in regulation of external acid-base balance are
the lungs are the kidneys.

The lungs are important for excretion of carbon dioxide (the respiratory
acid) and there is a huge amount of this to be excreted: at least 12,000 to
13,000 mmols/day.

In contrast the kidneys are responsible for excretion of the fixed acids and
this is also a critical role even though the amounts involved (70-100
mmols/day) are much smaller.

The main reason for this renal importance is because there is no other
way to excrete these acids and it should be appreciated that the amounts
involved are still very large when compared to the plasma [H +] of only 40
nanomoles/litre.

There is a second extremely important role that the kidneys play in acid-
base balance, namely the reabsorption of the filtered bicarbonate.
Bicarbonate is the predominant extracellular buffer against the fixed acids
and it important that its plasma concentration should be defended against
renal loss.
In acid-base balance, the kidney is responsible for 2 major activities:

 Reabsorption of filtered bicarbonate: 4,000 to 5,000 mmol/day

 Excretion of the fixed acids (acid anion and associated H +): about 1
mmol/kg/day.

Both these processes involve secretion of H + into the lumen by the renal
tubule cells but only the second leads to excretion of H + from the body.
The renal mechanisms involved in acid-base balance can be difficult to
understand so as a simplification we will consider the processes occurring
in the kidney as involving 2 aspects:

 Proximal tubular mechanism

 Distal tubular mechanism

PROXIMAL TUBULAR MECHANISM

The contributions of the proximal tubules to acid-base balance are:

 firstly, reabsorption of bicarbonate which is filtered at the glomerulus

 secondly, the production of ammonium

BICARBONATE REABSORPTION

Daily filtered bicarbonate equals the product of the daily glomerular

filtration rate (180 l/day) and the plasma bicarbonate concentration (24
mmol/l). This is 180 x 24 = 4320 mmols/day (or usually quoted as
between 4000 to 5000 mmols/day).

About 85 to 90% of the filtered bicarbonate is reabsorbed in the proximal

tubule and the rest is reabsorbed by the intercalated cells of the distal
tubule and collecting ducts.

The reactions that occur are outlined in the diagram. Effectively, H + and
HCO3- are formed from CO2 and H2O in a reaction catalysed by carbonic
anhydrase. The actual reaction involved is probably formation of H + and
OH- from water, then reaction of OH- with CO2 (catalysed by carbonic
anhydrase) to produce HCO3-. Either way, the end result is the same.
The H+ leaves the proximal tubule cell and enters the PCT lumen by 2
mechanisms:

 Via a Na+-H+ antiporter (major route)

 Via H+-ATPase (proton pump)

Filtered HCO3- cannot cross the apical membrane of the PCT cell.
Instead it combines with the secreted H+ (under the influence of brush
border carbonic anhydrase) to produce CO2 and H2O. The CO2 is lipid
soluble and easily crosses into the cytoplasm of the PCT cell. In the cell, it
combines with OH- to produce bicarbonate. The HCO3- crosses the
basolateral membrane via a Na+-HCO3- symporter. This symporter is
electrogenic as it transfers three HCO3- for every one Na+. In comparison,
the Na+-H+ antiporter in the apical membrane is not electrogenic because
an equal amount of charge is transferred in both directions.

The basolateral membrane also has an active Na +-K+ ATPase (sodium

pump) which transports 3 Na+ out per 2 K+ in. This pump is electrogenic in
a direction opposite to that of the Na+-HCO3- symporter. Also the sodium
pump keeps intracellular Na+ low which sets up the Na+ concentration
gradient required for the H+-Na+ antiport at the apical membrane. The H+-
Na+ antiport is an example of secondary active transport.
The net effect is the reabsorption of one molecule of HCO 3 and one
molecule of Na+ from the tubular lumen into the blood stream for each
molecule of H+ secreted. This mechanism does not lead to the net
excretion of any H+ from the body as the H+ is consumed in the reaction
with the filtered bicarbonate in the tubular lumen.

[Note: The differences in functional properties of the apical membrane

from that of the basolateral membranes should be noted. This difference
is maintained by the tight junctions which link adjacent proximal tubule
cells. These tight junctions have two extremely important functions:
Gate function: They limit access of luminal solutes to the intercellular
space. This resistance can be altered and this paracellular pathway can
be more open under some circumstances (ie the gate can be opened a
little).

Fence function: The junctions maintain different distributions of some of

the integral membrane proteins. For example they act as a fence to keep
the Na+-H+ antiporter limited to the apical membrane, and keep the Na +-
K+ ATPase limited to the basolateral membrane. The different distribution
of such proteins is absolutely essential for cell function.]

The 4 major factors which control bicarbonate reabsorption are:

 Luminal HCO3- concentration

 Luminal flow rate

 Arterial pCO2

 Angiotensin II (via decrease in cyclic AMP)

An increase in any of these four factors causes an increase in

bicarbonate reabsorption. Parathyroid hormone also has an effect: an
increase in hormone level increases cAMP and decreases bicarbonate
reabsorption.

The mechanism for H+ secretion in the proximal tubule is described as a

high capacity, low gradient system:

The high capacity refers to the large amount (4000 to 5000 mmols) of
H+ that is secreted per day. (The actual amount of H + secretion is 85% of
the filtered load of HCO3-).

The low gradient refers to the low pH gradient as tubular pH can be

decreased from 7.4 down to 6.7-7.0 only.

Though no net excretion of H+ from the body occurs, this proximal

mechanism is extremely important in acid-base balance. Loss of
bicarbonate is equivalent to an acidifying effect and the potential amounts
of bicarbonate lost if this mechanism fails are very large.

AMMONIUM PRODUCTION

Ammonium (NH4) is produced predominantly within the proximal tubular

cells. The major source is from glutamine which enters the cell from the
peritubular capillaries (80%) and the filtrate (20%). Ammonium is
produced from glutamine by the action of the enzyme glutaminase.
Further ammonium is produced when the glutamate is metabolised to
produce alpha-ketoglutarate. This molecule contains 2 negatively-charged
carboxylate groups so further metabolism of it in the cell results in the
production of 2 HCO3- anions. This occurs if it is oxidised to CO2 or if it is
metabolised to glucose.
The pKa for ammonium is so high (about 9.2) that both at extracellular
and at intracellular pH, it is present entirely in the acid form NH 4+. The
previous idea that lipid soluble NH 3 is produced in the tubular cell, diffuses
into the tubular fluid where it is converted to water soluble NH 4+ which is
now trapped in the tubule fluid is incorrect.
The subsequent situation with ammonium is complex. Most of the
ammonium is involved in cycling within the medulla. About 75% of the
proximally produced ammonium is removed from the tubular fluid in the
medulla so that the amount of ammonium entering the distal tubule is
small. The thick ascending limb of the loop of Henle is the important
segment for removing ammonium. Some of the interstitial ammonium
returns to the late proximal tubule and enters the medulla again (ie
recycling occurs).
An overview of the situation so far is that:

 The ammonium level in the DCT fluid is low because of removal in

the loop of Henle

 Ammonium levels in the medullary interstitium are high (and are kept
high by the recycling process via the thick ascending limb and the late
PCT)

 Tubule fluid entering the medullary collecting duct will have a low pH
if there is an acid load to be excreted (and the phosphate buffer has
been titrated down.

If H+ secretion continues into the medullary collecting duct this would

reduce the pH of the luminal fluid further. A low pH greatly augments
transfer of ammonium from the medullary interstitium into the luminal fluid
as it passes through the medulla. The lower the urine pH, the higher the
ammonium excretion and this ammonium excretion is augmented further
if an acidosis is present.

This augmentation with acidosis is 'regulatory' as the increased

ammonium excretion by the kidney tends to increase extracellular pH
towards normal.
If the ammonium returns to the blood stream it is metabolised in the liver
to urea (Krebs-Henseleit cycle) with net production of one hydrogen ion
per ammonium molecule.

DISTAL TUBULAR MECHANISM

This is a low capacity, high gradient system which accounts for the
excretion of the daily fixed acid load of 70 mmols/day. The maximal
capacity of this system is as much as 700 mmols/day but this is still low
compared to the capacity of the proximal tubular mechanism to secrete
H+.

It can however decrease the pH down to a limiting pH of about 4.5 : this

represents a thousand-fold (ie 3 pH units) gradient for H + across the distal
tubular cell. The maximal capacity of 700 mmols/day takes about 5 days
to reach.
The processes involved are:-

 Formation of titratable acidity (TA)

 Addition of ammonium (NH4+) to luminal fluid

 Reabsorption of Remaining Bicarbonate

Classification of Carbohydrates?
The main classification of carbohydrate is done on the basis of hydrolysis.
This classification is as follow:

1. Monosaccharides: These are the simplest form of carbohydrate that

cannot be hydrolyzed any further. They have the general formula of
(CH2O)n. Some common examples are glucose, Ribose etc.
2. Oligosaccharides: Carbohydrates that on hydrolysis yield two to ten
smaller units or monosaccharides are oligosaccharides. They are a
large category and further divides into various subcategories.

3. Disaccharides: A further classification of oligosaccharides, these give

two units of the same or different monosaccharides on hydrolysis. For
example, sucrose on hydrolysis gives one molecule of glucose and
fructose each. Whereas maltose on hydrolysis gives two molecules of
only glucose,

4. Trisaccharides: Carbohydrates that on hydrolysis gives three

molecules of monosaccharides, whether same or different. An example
is Raffinose.

5. Tetrasaccharides: And as the name suggests this carbohydrate on

hydrolysis give four molecules of monosaccharides. Stachyose is an
example.

6. Polysaccharides: The final category of carbohydrates. These give a

large number of monosaccharides when they undergo hydrolysis, These
carbohydrates are not sweet in taste and are also known as non-sugars.
Some common examples are starch, glycogen etc.

Oxidation of fatty acid?

Triacylglycerols (fats) are the most abundant source of energy and

provide energy twice as much as carbohydrates and proteins. This is
achieved because the fatty acids which are present in the triacylglycerols
are already in the reduced form. To convert fats into energy, the digested
fats or the stored fats have to be first activated and transported to the
mitochondrial matrix as all the enzymes required for metabolism
(oxidation) are present there.

For activation of fatty acids, they are converted to fatty acyl-CoA by the
enzyme thiokinase in the cytosol and then with the help of carnitine (L-
carnitine is the active stereoisomer) is transported through the
mitochondrial membranes into the mitochondrial matrix.
Oxidation of fatty acids occurs in three stages:

 β-oxidation of fatty acids resulting in cleavage of two-carbon units (α

and β carbons) from the carboxyl end of fatty acyl-CoA with the
formation of acetyl CoA. This reaction keeps occuring till the entire
fatty acyl chain is broken down to acetyl CoA molecules. For eg.
Palmitoyl CoA (16 carbon chain) on β-oxidation will give eight acetyl
CoA molecules. It is further discussed in detail below.
 The acetyl groups produced from β-oxidation of the fatty acid
participate in the Kreb’s cycle resulting in the formation of NADH and
FADH2.
 The reduced coenzymes (NADH and FADH2) are oxidized by giving
up the protons and electrons to oxygen present in the mitochondria to
synthesize ATP by oxidative phosphorylation in the Electron
Transport System.

Beta oxidation of Fatty Acids

Once the fatty acids have been transported to the mitochondrial matrix via
carnitine pathway, β-oxidation of fatty acyl-CoA (n carbons) occurs
within the mitochondria in four steps as discussed below:

 First step– Fatty acyl-CoA is acted upon by an enzyme acyl-CoA

dehydrogenase which is FAD dependent. Fatty acyl-
CoAundergoes dehydrogenation and forms a trans-double bond at
the α and β carbons to form trans-Δ2-enoyl-CoA. Acyl-CoA
dehydrogenase are present as three isoenzymes each specific for a
particular carbon chain length (short, intermediate and long). The
electrons which were removed from the fatty acyl-CoA chain are
transferred to FAD which gets reduced to FADH2. This
FADH2 immediately via the Electron Transport System gets converted
to ATP molecules.
 Second step– Enoyl-CoA hydratase catalyzes this reaction
where water is added. Hydration occurs at the double bond resulting
in the formation of β-hydroxyacyl-CoA.
 Third step– β-hydroxyacyl-CoA undergoes dehydrogenation to
form β-ketoacyl-CoA in the presence of β-hydroxyacyl-CoA
dehydrogenase. The electrons available as a result of
dehydrogenation are accepted by NAD+ to form NADH + H+ which
 immediately exchanges these electrons with oxygen in the Electron
Transport System to form ATP molecules.
 Fourth step– This reaction is called as thiolysis as acyl-CoA
acetyltransferase(also known as thiolase) in the presence of CoA-
SH causes the cleavage of β-ketoacyl-CoA to form acetyl CoA and
the thioester of the original fatty acid with two carbons less. This
cleavage occurs as the β carbon ketone group is a good target for
nucleophilic attack by the thiol (-SH) group of the coenzyme A.

The new fatty acyl-CoA (n-2 carbons) formed again participates in the β-
oxidation cycle to form a new fatty acyl-CoA with two carbons less (n-4
carbons) and a new molecule of acetyl CoA. This process continues till the
entire fatty acid is converted into acetyl CoA molecules.
Acetyl CoA formed from the above steps now enters the Kreb’s cycle to
get oxidized to CO2 and H2O.

Note:
 Oxidation of odd carbon length fatty acids has additional reactions
and is described in detail here: Oxidation of odd carbon chain
length fatty acids.
 For unsaturated fatty acids β-oxidation has two more additional steps
are required catalyzed by two enzymes isomerase and reductase.
Oxidation of unsaturated fats gives less energy as compared to
saturated fats. Read more about it here: Oxidation of unsaturated
fatty acids.
Classification of Protein?
Based on Chemical composition
On the basis of their chemical composition, proteins may be divided into
two classes: simple and complex.

1. Simple Protein

Also known as homoproteins, they are made up of only amino acids.

Examples are plasma albumin, collagen, and keratin.

2. Conjugated Protein

Sometimes also called heteroproteins, they contain in their structure

a non-protein portion. Three examples are glycoproteins,
chromoproteins, and phosphoproteins.

Glycoprotein

 They are proteins that covalently bind one or more carbohydrate units
to the polypeptide backbone.
Typically, the branches consist of not more than 15-
20 carbohydrate units, where you can find arabinose, fucose (6-
deoxygalactose), galactose, glucose, mannose, N-acetylglucosamine
(GlcNAc, or NAG), and N-acetylneuraminic acid (Neu5Ac or NANA).

Examples of glycoproteins are:

glycophorin, the best known among erythrocyte membrane glycoproteins;
fibronectin, that anchors cells to the extracellular matrix through
interactions on one side with collagen or other fibrous proteins, while on the
other side with cell membranes;
all blood plasma proteins, except albumin;
immunoglobulins or antibodies.

Chromoprotein
 They are proteins that contain colored prosthetic groups.
Typical examples are:
hemoglobin and myoglobin, which bind, respectively, one and four heme
groups;
chlorophylls, which bind a porphyrin ring with a magnesium atom at its
centre;
rhodopsins, which bind retinal.
Phosphoprotein
They are proteins that bind phosphoric acid to serine and threonine
residues.
Generally, they have a structural function, such as tooth dentin, or reserve
function, such as milk caseins (alpha, beta, gamma and delta), and egg
yolk phosvitin.

CLASSIFICATION BASED ON SHAPE

On the basis of their shape, proteins may be divided into two classes:
fibrous and globular.

1. Fibrous protein
They have primarily mechanical and structural functions, providing
support to the cells as well as the whole organism.
These proteins are insoluble in water as they contain, both internally and
on their surface, many hydrophobic amino acids. The presence on their
surface of hydrophobic amino acids facilitates their packaging into very
complex supramolecular structures.

In this regard, it should be noted that their polypeptide chains form long
filaments or sheets, where in most cases only one type of secondary
structure, that repeats itself, is found.
In vertebrates, these proteins provide external protection, support and
shape; in fact, thanks to their structural properties, they ensure flexibility
and/or strength.

Some fibrous proteins, such as α-keratins, are only partially hydrolyzed in

the intestine.
Here are some examples.

Fibroin

It is produced by spiders and insects. An example is that produced by the

silkworm, Bombyx mori.

Collagen
The term “collagen” indicates not a single protein but a family of
structurally related proteins (at least 29 different types), which constitute
the main protein component of connective tissue, and more generally, the
extracellular scaffolding of multicellular organisms. In vertebrates, they
represent about 25-30% of all proteins.

They are found in different tissues and organs, such as tendons and the
organic matrix of bone, where they are present in very high percentages,
but also in cartilage and in the cornea of the eye.

In the different tissues, they form different structures, each capable of

satisfying a particular need. For example, in the cornea, the molecules are
arranged in an almost crystalline array, so that they are virtually
transparent, while in the skin they form fibers not very intertwined and
directed in all directions, which ensure the tensile strength of the skin itself.
Note: the different types of collagen have low nutritional value as deficient
in several amino acids (in fact, they contain no tryptophan and low amount
of the other essential amino acids).

The gelatin used in food preparation is a derivative of collagen.

Alpha-keratin

They constitute almost the entire dry weight of nails, claws, beak, hooves,
horns, hair, wool, and a large part of the outer layer of the skin.
The different stiffness and flexibility of these structures is a consequence
of the number of disulfide bonds that contribute, together with other
binding forces, to stabilize the proteinstructure. And this is the reason why
wool keratins, which have a low number of disulfide bonds, are flexible,
soft and extensible, unlike claw and beak keratins that are rich in disulfide
bonds.

Elastin
This protein provides elasticity to the skin and blood vessels, a
consequence of its random coiled structure, that differs from the structures
of the α-keratins and collagens.

2. Globular Protein

Most of the proteins belong to this class.

They have a compact and more or less spherical structure, more
complex than fibrous proteins. In this regard, motifs, domains, tertiary and
quaternary structures are found, in addition to the secondary structures.
They are generally soluble in water but can also be found inserted into
biological membranes (transmembrane proteins), thus in a hydrophobic
environment.
Unlike fibrous proteins, that have structural and mechanical functions, they
act as:

 enzymes;
 hormones;
 membrane transporters and receptors;
 transporters of triglycerides, fatty acids and oxygen in the blood;
 immunoglobulins or antibodies;
 grain and legume storage proteins.
Examples of globular proteins are myoglobin, hemoglobin, and cytochrome
c.
At the intestinal level, most of the globular proteins of animal origin are
hydrolyzed almost entirely to amino acids.

CLASSIFICATION BASED ON BIOLOGICAL FUNCTION

The multitude of functions that proteins perform is the consequence of both

the folding of the polypeptide chain, therefore of their three-dimensional
structure, and the presence of many different functional groups in the
amino acid side chains, such as thiols, alcohols, thioethers, carboxamides,
carboxylic acids and different basic groups.

From the functional point of view, they may be divided into several groups.
 Enzymes (biochemical catalysts).
In living organisms, almost all reactions are catalyzed by
specific proteins called enzymes. They have a high catalytic power,
increasing the rate of the reaction in which they are involved at least by
 factor 106. Therefore, life as we know could not exist without their
“facilitating action”.
Almost all known enzymes, and in the human body they are thousand,
are proteins (except some catalytic RNA molecules called ribozymes,
that is, ribonucleic acid enzymes).

 Transport proteins
Many small molecules, organic and inorganic, are transported in the
bloodstream and extracellular fluids, across the cell membranes, and
inside the cells from one compartment to another, by specific proteins.
Examples are:
 hemoglobin, that carries oxygen from the alveolar blood vessels to
tissue capillaries;
transferrin, which carries iron in the blood;
membrane carriers;
fatty acid binding proteins (FABP), that is, the proteinsinvolved in the
intracellular transport of fatty acids;
proteins of plasma lipoproteins, macromolecular complexes
of proteins and lipids responsible for the transport of triglycerides,
which are otherwise insoluble in water;
albumin, that carries free fatty acids, bilirubin, thyroid hormones, and
certain medications such as aspirin and penicillin, in the blood.
 Many of these proteins also play a protective role, since the bound
molecules, such as fatty acids, may be harmful for the organism
when present in free form.
 Storage proteins
Examples are:
ferritin, that stores iron intracellularly in a non-toxic form;
milk caseins, that act as a reserve of amino acids for the milk;
egg yolk phosvitin, that contains high amounts of phosphorus;
prolamins and glutelins, the storage proteins of cereals.
 Mechanical support
Proteins have a pivotal role in the stabilization of many structures.
Examples are α-keratins, collagen and elastin. The same cytoskeletal
system, the scaffold of the cell, is made of proteins.
 They generate movement.
They are responsible, among others, for:
the contraction of the muscle fibers (of which myosin is the main
component);
the propulsion of spermatozoa and microorganisms with flagella;
the separation of chromosomes during mitosis.

 They are involved in nerve transmission.

An example is the receptor for acetylcholine at synapses.
 They control development and differentiation.
Some proteins are involved in the regulation of gene expression. An
example is the nerve growth factor (NGF), discovered by Rita Levi-
Montalcini, that plays a leading role in the formation of neural networks.
 Hormones
Many hormones are proteins.
They are regulatory molecules involved in the control of many cellular
functions, from metabolism to reproduction. Examples are insulin,
glucagon, and thyroid-stimulating hormone (TSH).
 Protection against harmful agents.
The antibodies or immunoglobulins are glycoproteins that recognize
antigens expressed on the surface of viruses, bacteria and other
infectious agents.
Interferon, fibrinogen, and factors of blood coagulation are other
members of this group.
 Storage of energy.
Proteins, and in particular the amino acids that constitute them, act as
energy storage, second in size only to the adipose tissue, that in
particular conditions, such as prolonged fasting, may become essential
for survival. However, their reduction of more than 30% leads to a
decrease of the contraction capacity of respiratory muscle, immune
function, and organ function, that are not compatible with life.
Therefore, proteins are an extremely valuable fuel.

CLASSIFICATION BASED ON SOLUBILITY

The different globular proteins can be classified based on their solubility

in different solvents, such as water, salt and alcohol
 Biosynthesis of Triacyl Glycerol?
Three main pathways for triacylglycerol biosynthesis are known,
the sn-glycerol-3-phosphate and dihydroxyacetone phosphate
pathways, which predominates in liver and adipose tissue, and a
monoacylglycerol pathway in the intestines.

The most important route to triacylglycerols is the sn-glycerol-3-

phosphate or Kennedy pathway, first described by Professor
Eugene Kennedy and colleagues in the 1950s, by means of which
more than 90% of liver triacylglycerols are produced.
In this pathway, the main source of the glycerol backbone has long been
believed to be sn-glycerol-3-phosphate produced by the catabolism of
glucose (glycolysis) or to a lesser extent by the action of the enzyme
glycerol kinase on free glycerol. However, there is increasing evidence that
a significant proportion of the glycerol is produced de novo by a process
known as glyceroneogenesis via pyruvate. Indeed, this may be the main
source in adipose tissue.
Subsequent reactions occur primarily in the endoplasmic reticulum. First,
the precursor sn-glycerol-3-phosphate is esterified by a fatty acid
coenzyme A ester in a reaction catalysed by a glycerol-3-phosphate
acyltransferase (GPAT) at position sn-1 to form lysophosphatidic acid, and
this is in turn acylated by an acylglycerophosphate acyltransferase
(AGPAT) in position sn-2 to form a key intermediate in the biosynthesis of
all glycerolipids - phosphatidic acid. Numerous isoforms of these
enzymes are known; they are expressed with specific tissue and
membrane distributions and they are regulated in different ways.

The phosphate group is removed by an enzyme (or family of enzymes)

phosphatidic acid phosphohydrolase (PAP or ‘phosphatidate phosphatase’
or ‘lipid phosphate phosphatase’). PAP is also important as it produces sn-
1,2-diacylglycerols as essential intermediates in the biosynthesis not only
of triacylglycerols but also of phosphatidylcholine and
phosphatidylethanolamine (and of monogalactosyldiacylglycerols in plants).
In contrast to the activity responsible for phospholipid biosynthesis in
mammals, much of the phosphatase activity leading to triacylglycerol
biosynthesis resides in three related cytoplasmic proteins, termed lipin-1,
lipin-2 and lipin-3, which were characterized before the nature of their
enzymatic activities were determined.

The lipins are tissue specific, and each appears to have distinctive
expression and functions, but lipin-1 (PAP1) in three isoforms (designated
1α, 1β and 1γ) accounts for all the PAP activity in adipose tissue and
skeletal muscle.

While it occurs mainly in the cytosolic compartment of cells, it is

translocated to the endoplasmic reticulum in response to elevated levels of
fatty acids within cells although it does not have trans-membrane domains.
Lipin-1 activity requires Mg2+ ions and is inhibited by N-ethylmaleimide,
whereas the membrane-bound activity responsible for synthesising
diacylglycerols as a phospholipid intermediate is independent of
Mg2+ concentration and is not sensitive to the inhibitor.

Perhaps surprisingly, lipin-1 has a dual role in that it operates in

collaboration with known nuclear receptors as a transcriptional coactivator
to modulate lipid metabolism (lipin 1α) while lipin 1β is associated with
induction of lipogenic genes such as fatty acid synthase, stearoyl CoA
desaturase and DGAT. Abnormalities in lipin-1 expression are known to be
involved in some human disease states that may lead to the metabolic
syndrome. Lipin 2 is a similar phosphatidate phosphohydrolase, which is
present in liver and brain and is regulated dynamically by fasting and
obesity (in mice), while lipin 3 is found in the gastrointestinal tract and liver.

In the final step in this pathway, the resultant 1,2-diacyl-sn-glycerol is

acylated by diacylglycerol acyltransferases (DGAT), which can utilize a
wide range of fatty acyl-CoA esters to form the triacyl-sn-glycerol. In fact
there are two DGAT enzymes, which are structurally and functionally
distinct. In animals, DGAT1 is located mainly in the endoplasmic reticulum
and is expressed in skeletal muscle, skin and intestine, with lower levels of
expression in liver and adipose tissue.

It is believed to have dual topology contributing to triacylglycerol synthesis

on both sides of the membrane of the endoplasmic reticulum, but
esterifying only pre-formed fatty acids of exogenous origin. Perhaps
surprisingly, DGAT1 is the only one present in the epithelial cells that
synthesise milk fat in the mammary gland. Orthologues of this enzyme are
present in most eukaryotes, other than yeasts, and it is especially important
in plants. Also DGAT1 can utilize a wider range of substrates, including
monoacylglycerols, long-chain alcohols (for wax synthesis) and retinol.
DGAT2 is the main form of the enzyme in hepatocytes and adipocytes (lipid
droplets), although it is expressed much more widely in tissues. It is
associated with distinct regions of the endoplasmic reticulum, at the surface
of lipid droplets and in mitochondria, and it esterifies fatty acids of both
endogenous and exogenous origin.

DGAT2 is believed to have a targeting domain that enables it to tether

between the endoplasmic reticulum and lipid droplet thereby channelling
triacylglycerols from the synthesis site in the endoplasmic reticulum to the
nascent lipid droplet, where they accumulate and lead to the expansion of
the latter. Both enzymes are important modulators of energy metabolism,
although DGAT2 appears to be especially important in controlling the
homeostasis of triacylglycerols in vivo. As the glycerol-3-phosphate
acyltransferase (GPAT) has the lowest specific activity of these enzymes,
this step may be the rate-limiting one. However, DGATs are the dedicated
triacylglycerol-forming enzymes, and they are seen as the best target for
pharmaceutical intervention in obesity and attendant ailments; clinical
studies of DGAT1 inhibitors are at an early stage.

Different types of lipoprotein and its

function?
Lipoprotein, any member of a group of substances containing
both lipid (fat) and protein. They occur in both soluble complexes—as
in egg yolk and mammalian blood plasma—and insoluble ones, as
in cell membranes. Lipoproteins in blood plasma have been
intensively studied because they are the mode of transport
for cholesterol through the bloodstream and lymphatic fluid.

Lipids are transported in the circulation packaged in lipoproteins. The

clinical relevance of blood lipid levels is that abnormal levels of lipids in
certain lipoproteins are linked to an increase risk of atherosclerosis.
Atherosclerosis is a cardiovascular disease in which lipids and
inflammatory cells accumulate in plaques within the walls of blood vessels.
As a result, vessel walls are narrowed and clots may form, impeding blood
flow and oxygen delivery and causing tissue injury. Heart disease occurs
because the coronary arteries supplying the heart are a major site where
atherosclerotic plaques form.

The liver is central to the regulation of cholesterol levels in the body. Not
only does it synthesize cholesterol for export to other cells, but it also
removes cholesterol from the body by converting it to bile salts and putting
it into the bile where it can be eliminated in the feces. Furthermore, the liver
synthesizes the various lipoproteins involved in transporting cholesterol and
other lipids throughout the body.

Cholesterol synthesis in the liver is under negative feedback regulation.

Increased cholesterol in a hepatocyte leads to decreased activity of HMG-
CoA reductase, the rate-limiting enzyme in cholesterol synthesis.

TYPES OF LIPOPROTEINS

Lipoproteins are particles that contain triacylglycerol (TAG), cholesterol,

phospholipids and amphipathic proteins called apolipoproteins. (You can
refresh your memory about the structure of lipoproteins by visiting the
webpage Lipoproteins from fall quarter). Lipoproteins can be differentiated
on the basis of their density, but also by the types of apolipoproteins they
contain. The degree of lipid in a lipoprotein affects its density—the lower
the density of a lipoprotein, the more lipid it contains relative to protein. The
four major types of lipoproteins are chylomicrons, very low-density
lipoprotein (VLDL), low-density lipoprotein (LDL), and high-density
lipoprotein (HDL).
 Chylomicrons and VLDL deliver TAG to cells in the body. Two
types of lipoproteins are triglyceride-rich: the chylomicrons and VLDL.
Chylomicrons are synthesized by enterocytes from lipids absorbed in
the small intestine. VLDL is synthesized in the liver. The function of
these lipoproteins is to deliver energy-rich triacylglycerol (TAG) to
cells in the body (pink pathway). TAG is stripped from chylomicrons
and VLDL through the action of lipoprotein lipase, an enzyme that is
found on the surface of endothelial cells. This enzyme digests the
TAG to fatty acids and monoglycerides, which can then diffuse into
the cell to be oxidized, or in the case of an adipose cell, to be re-
synthesized into TAG and stored in the cell.

 LDL delivers cholesterol to cells in the body. As VLDL particles

are stripped of triacylglycerol, they become more dense. These
particles are remodeled at the liver and transformed into LDL. The
function of LDL is to deliver cholesterol to cells, where it is used in
membranes, or for the synthesis of steroid hormones (blue pathway).
Cells take up cholesterol by receptor-mediated endocytosis. LDL
binds to a specific LDL receptor and is internalized in an endocytic
 vesicle. Receptors are recycled to the cell surface, while hydrolysis in
an endolysosome releases cholesterol for use in the cell.

 HDL is involved in reverse cholesterol transport. Excess

cholesterol is eliminated from the body via the liver, which secretes
cholesterol in bile or converts it to bile salts. The liver removes LDL
and other lipoproteins from the circulation by receptor-mediated
endocytosis. Additionally, excess cholesterol from cells is brought
back to the liver by HDL in a process known as reverse cholesterol
transport (green pathway). HDL (or really, the HDL precursor) is
synthesized and secreted by the liver and small intestine. It travels in
the circulation where it gathers cholesterol to form mature HDL, which
then returns the cholesterol to the liver via various pathways.

Factors affecting or influencing Enzyme

Activity?
Knowledge of basic enzyme kinetic theory is important in enzyme
analysis in order both to understand the basic enzymatic mechanism
and to select a method for enzyme analysis.

The conditions selected to measure the activity of an enzyme would

not be the same as those selected to measure the concentration of its
substrate. Several factors affect the rate at which enzymatic reactions
proceed - temperature, pH, enzyme concentration, substrate
concentration, and the presence of any inhibitors or activators.

Enzyme Concentration
In order to study the effect of increasing the enzyme concentration upon
the reaction rate, the substrate must be present in an excess amount; i.e.,
the reaction must be independent of the substrate concentration. Any
change in the amount of product formed over a specified period of time will
be dependent upon the level of enzyme present.
 The amount of enzyme present in a reaction is measured by the
activity it catalyzes. The relationship between activity and
concentration is affected by many factors such as temperature, pH,
etc. An enzyme assay must be designed so that the observed activity
is proportional to the amount of enzyme present in order that the
enzyme concentration is the only limiting factor. It is satisfied only
when the reaction is zero order.

Substrate Concentration
It has been shown experimentally that if the amount of the enzyme is kept
constant and the substrate concentration is then gradually increased, the
reaction velocity will increase until it reaches a maximum. After this point,
increases in substrate concentration will not increase the velocity (delta
A/delta T). This is represented graphically in Figure 8.
Effects of Inhibitors on Enzyme Activity
Enzyme inhibitors are substances which alter the catalytic action of the
enzyme and consequently slow down, or in some cases, stop catalysis.
There are three common types of enzyme inhibition - competitive, non-
competitive and substrate inhibition.
Most theories concerning inhibition mechanisms are based on the
existence of the enzyme-substrate complex ES. As mentioned earlier, the
existence of temporary ES structures has been verified in the laboratory.
Competitive inhibition occurs when the substrate and a substance
resembling the substrate are both added to the enzyme. A theory called the
"lock-key theory" of enzyme catalysts can be used to explain why inhibition
occurs.
The lock and key theory utilizes the concept of an "active site." The concept
holds that one particular portion of the enzyme surface has a strong affinity
for the substrate. The substrate is held in such a way that its conversion to
the reaction products is more favorable. If we consider the enzyme as the
lock and the substrate the key (Figure 9) - the key is inserted in the lock, is
turned, and the door is opened and the reaction proceeds. However, when
an inhibitor which resembles the substrate is present, it will compete with
the substrate for the position in the enzyme lock. When the inhibitor wins, it
gains the lock position but is unable to open the lock. Hence, the observed
reaction is slowed down because some of the available enzyme sites are
occupied by the inhibitor. If a dissimilar substance which does not fit the
site is present, the enzyme rejects it, accepts the substrate, and the
reaction proceeds normally.
Non-competitive inhibitors are considered to be substances which when
added to the enzyme alter the enzyme in a way that it cannot accept the
substrate. Figure 10.
Substrate inhibition will sometimes occur when excessive amounts of
substrate are present. Figure 11 shows the reaction velocity
decreasing after the maximum velocity has been reached.

Additional amounts of substrate added to the reaction mixture after

this point actually decrease the reaction rate. This is thought to be
due to the fact that there are so many substrate molecules competing
for the active sites on the enzyme surfaces that they block the sites
(Figure 12) and prevent any other substrate molecules from
occupying them.
 This causes the reaction rate to drop since all of the enzyme present
is not being used.

Temperature Effects

Like most chemical reactions, the rate of an enzyme-catalyzed reaction

increases as the temperature is raised. A ten degree Centigrade rise in
temperature will increase the activity of most enzymes by 50 to 100%.
Variations in reaction temperature as small as 1 or 2 degrees may
introduce changes of 10 to 20% in the results. In the case of enzymatic
reactions, this is complicated by the fact that many enzymes are adversely
affected by high temperatures. As shown in Figure 13, the reaction rate
increases with temperature to a maximum level, then abruptly declines with
further increase of temperature. Because most animal enzymes rapidly
become denatured at temperatures above 40°C, most enzyme
determinations are carried out somewhat below that temperature.
Over a period of time, enzymes will be deactivated at even moderate
temperatures. Storage of enzymes at 5°C or below is generally the most
suitable. Some enzymes lose their activity when frozen.

EFFECTS of pH
Enzymes are affected by changes in pH. The most favorable pH value - the
point where the enzyme is most active - is known as the optimum pH. This
is graphically illustrated in Figure 14.

Extremely high or low pH values generally result in complete loss of

activity for most enzymes. pH is also a factor in the stability
of enzymes. As with activity, for each enzyme there is also a region of
pH optimal stability.
 Classify enzymes with examples?

There were six classes of enzymes that were created so that

enzymes could easily be named. These classes are:
Oxidoreductases, Transferases, Hydrolases, Lyases, Isomerases,
and Ligases. This is the international classification used for enzymes.
Enzymes are normally used for catalyzing the transfer of functional
groups, electrons, or atoms. Since this is the case, they are assigned
names by the type of reaction they catalyze.

Oxidoreductases catalyze oxidation-reduction reactions where

electrons are transferred. These electrons are usually in the form of
hydride ions or hydrogen atoms. When a substrate is being oxidized it
is the hydrogen donor. The most common name used is a
dehydrogenase and sometimes reductase will be used. An oxidase is
referred to when the oxygen atom is the acceptor.

Hydrolases catalyze reactions that involve hydrolysis. This cases

usually involves the transfer of functional groups to water. When the
hydrolase acts on amide, glycosyl, peptide, ester, or other bonds,
they not only catalyze the hydrolytic removal of a group from the
substrate but also a transfer of the group to an acceptor compound.
These enzymes could also be classified under transferases since
hydrolysis can be viewed as a transfer of a functional group to water
as an acceptor. However, as the acceptor's reaction with water was
discovered very early, it's considered the main function of the enzyme
which allows it to fall under this classification. For example:
Chymotrypsin.
Lyases catalyze reactions where functional groups are added to break
double bonds in molecules or the reverse where double bonds are formed
by the removal of functional groups. For example: Fructose bisphosphate
aldolase used in converting fructose 1,6-bisphospate to G3P and DHAP by
cutting C-C bond.
Isomerases catalyze reactions that transfer functional groups within a
molecule so that isomeric forms are produced. These enzymes allow for
structural or geometric changes within a compound. Sometime the
interconversion is carried out by an intramolecular oxidoreduction. In this
case, one molecule is both the hydrogen acceptor and donor, so there's no
oxidized product.
The lack of a oxidized product is the reason this enzyme falls under this
classification. The subclasses are created under this category by the type
of isomerism. For example: phosphoglucose isomerase for converting
glucose 6-phosphate to fructose 6-phosphate. Moving chemical group
inside same substrate.
Ligases are used in catalysis where two substrates are ligated and the
formation of carbon-carbon, carbon-sulfide, carbon-nitrogen, and carbon-
oxygen bonds due to condensation reactions. These reactions are couple
to the cleavage of ATP.
Translocase are enzymes that catalyze the movement of ions or
molecules across membranes or their separation within membranes. It is a
general term for a protein that assists in moving another molecule, usually
across a cell membrane. The reaction is designated as a transfer from
“side 1” to “side 2” because the designations “in” and “out”, which had
previously been used, can be ambiguous. Translocases are the most
common secretion system in Gram positive bacteria.

Absorption,transport and storage of iron?

Absorption of Iron

Absorption of iron is one of the first steps in iron metabolism. Metabolism

is a process of chemical interactions that generate energy from food that
you eat. Iron metabolism is the part of the process that manages iron in
the body. Abnormal iron metabolism can result in too much or too little iron
in the body, which can cause poor health or even death.

Iron enters the stomach where it is exposed to stomach acid and changed
into a form that allows it to be absorbed. The portion of the small intestine
called the duodenum is the chief area where iron absorption takes place.
There may be a second minor absorption site near the end of the small
intestinal tract.

Once iron is absorbed it is carried (transported) by a protein called

transferrin. Each molecule of transferrin can carry two atoms of iron. When
working normally, transferrin binds to iron, and transports it to all tissues,
vital organs, and bone marrow, so that normal metabolism, DNA synthesis,
and red blood cell production can take place. Recently scientists have
discovered that transferrin does not work completely alone in the transport
of iron. Ceruloplasmin a major copper-containing protein in plasma is also
involved in iron transport. Iron needs adequate amounts of copper to reach
some of its intended destinations, such as the brain.

Transferrin is the major transporter of iron and ideally should be about 25-
35% saturated with iron.

Transferrin molecules that are heavily loaded (saturated) lose the ability to
hold onto (bind) iron. Unbound or free iron is highly destructive and
dangerous. Unbound iron can trigger free radical activity, which can cause
cell death, and destroy DNA. Unbound iron is sometimes called
uncontrolled iron.

One place that transferrin carries iron to is ferritin. Ferritin is a protein that
acts like a large holding vessel. Ferritin contains iron that we don't presently
need. It is sometimes called an iron storage protein. Ferritin is produced by
nearly every cell of the body. The brain contains huge amounts of ferritin,
so does the liver. Ferritin is a very large molecule; one ferritin molecule
alone can hold up to 4, 500 atoms of iron.

Elevated serum ferritin can be a sign that the person has inflammation due
to disease, or that potential disease causing factors such as iron overload
may be present.

Like transferrin, ferritin can also become unstable, and ineffective. Think of
ferritin like a big sink; when this sink gets full, ferritin and its iron can be
changed into something called hemosiderin.
Hemosiderin is a yellowish-brown substance that contains ferric oxide
(rust). A small amount of hemosiderin in tissues is probably normal and
may not be harmful, but when large amounts of the substance is allowed to
collect in organs, it then becomes a threat to good health. Hemosiderin can
accumulate in cells of the heart, liver, lungs, pancreas, central nervous
system, thyroid, reproductive organs, skin, adrenals, pituitary and thyroid
gland.

When the build up of hemosiderin is great, the organ cannot function

properly. For example, when beta cells (insulin producing cells of the
pancreas) are loaded with hemosiderin, these cells become unable to
produce or store adequate amounts of the hormone insulin, which results in
diabetes.

IRON THAT IS NOT ABSORBED:

For those with normal iron metabolism, unabsorbed iron, about 90% of iron
ingested through diet and supplements, is taken up by specific cells in the
intestinal tract, called enterocytes. These cells become engorged with iron,
die, drop off, and are excreted in feces.

IRON TRANSPORT

Iron in intestinal mucosal cells or stored in the liver may be transferred into
the blood for transport to other tissues.

The iron (III) storage form must be reduced to iron (II) in order to cross the
plasma membrane.

In the blood, iron (II) is reoxidized to iron (III) by ferroxidase II.

Iron (III) is carried by serum protein, transferrin.

Transferrin contains two sites that bind iron (III) tightly.

Normally

 about 1/9 of the transferrin molecules have iron bound at both sites
  about 4/9 of them have iron bound at one site

 and about 4/9 have no iron bound.

This means that transferrin is normally only about 1/3 saturated with iron
(the summary of saturation in the list above indicates that about six out of
every 18 sites are occupied), and there is a substantial unsaturated plasma
iron binding capacity. An unexpected influx of iron can be handled easily.

The iron binding capacity of serum is of clinical interest. It is accounted for

almost entirely by transferrin.

There are three components to the iron binding capacity of serum

 Serum iron is the concentration of iron present. Normally it is about

100 micrograms of iron per 100 milliliters of blood.

 Total iron binding capacity (TIBC) is the maximum amount of iron that
can be bound. Normally this is about 300 micrograms per 100
milliliters.

 The unsaturated iron binding capacity (UIBC) is the difference

between the TIBC and the serum iron. It is normally about 200.

Iron binding capacity is used in the differential diagnosis of certain

diseases

 In conditions associated with increased need for iron (iron deficiency

or late pregnancy) TIBC is increased, but saturation is decreased
from the normal 33%.

 In hemochromatosis, TIBC is low, but it is saturated.

 Certain other clinical conditions are associated with their own

characteristic patterns of TIBC and percent saturation.
IRON STORAGE

Iron is stored, mostly in the liver, as ferritin or hemosiderin.

Ferritin is a protein with a capacity of about 4500 iron (III) ions per protein
molecule. This is the major form of iron storage.

If the capacity for storage of iron in ferritin is exceeded, a complex of iron

with phosphate and hydroxide forms. This is called hemosiderin; it is
physiologically available.

As the body burden of iron increases beyond normal levels, excess

hemosiderin is deposited in the liver and heart. This can reach the point
that the function of these organs is impaired, and death ensues.

Several conditions can lead to excess body iron.

 Idiopathic hemochromatosis is a condition in which control of iron

absorption is defective, and excess iron is absorbed.

 Multiple transfusions can also lead to an excess body burden of iron.

This is a serious problem for persons with beta-thalassemia, a
disease in which hemoglobin is not made normally, and is supplied by
blood transfusion as needed.

Treatment of excess iron storage involves artificial removal of iron

from the body.

 Bleeding is the treatment of choice for idiopathic hemochromatosis.

 For beta-thalassemia bleeding would be inappropriate since the

disease consists of inability of the body to synthesize a blood
component, hemoglobin. In this case chelators are administered
which bind iron. The complex of chelator with iron is excreted in the
urine.

o A commonly used chelator is deferoxamine.

 o

o Although it is effective, it must be administered by an

inconvenient process of overnight infusion.

o More unfortunately, treatment becomes critical as the patients

reach their teenage years, a period of life associated with
rebelliousness and feelings of invulnerability. These patients
tend to be unreceptive to the treatment.

Biochemical function of vitamin A?

Vitamin A plays a role in a variety of functions throughout the body, such as:

 Vision

 Gene transcription

 Immune function

 Embryonic development and reproduction

 Bone metabolism

 Haematopoiesis

 Skin and cellular health

 Teeth

 Mucous membrane
Vision
 The role of vitamin A in the visual cycle is specifically related to the
retinal form. Rhodopsin is needed to see in low light (contrast) as well as
for night vision. Kühne showed that rhodopsin in the retina is only
regenerated when the retina is attached to retinal pigmented
epithelium, which provides retinal.
It is for this reason that a deficiency in vitamin A will inhibit the
reformation of rhodopsin and lead to one of the first symptoms, night
blindness.

Gene transcription
Vitamin A, in the retinoic acid form, plays an important role in gene
transcription. Once retinol has been taken up by a cell, it can be
oxidized to retinal (retinaldehyde) by retinol dehydrogenases and then
retinaldehyde can be oxidized to retinoic acid by retinaldehyde
dehydrogenases

Immune function
Vitamin A plays a role in many areas of the immune system, particularly in
T cell differentiation and proliferation. Hematopoietic stem cells are
important for the production of all blood cells, including immune cells, and
are able to replenish these cells throughout the life of an individual. Vitamin
A is important for the correct regulation of hematopoietic stem cell
dormancy.
Vitamin A has also been shown to be important for T cell homing to the
intestine, effects dendritic cells, and can play a role in
increased IgA secretion which is important for the immune response in
mucosal tissues.

Dermatology
Vitamin A, and more specifically, retinoic acid, appears to maintain normal
skin health by switching on genes and differentiating keratinocytes
(immature skin cells) into mature epidermal cells.
HOMOPOLYSACCHARIDE?

Homopolysaccharides are polysaccharides composed of a single type of

sugar monomer. For example, cellulose is an unbranched
homopolysaccharide made up of glucose monomers connected via beta-
glycosidic linkages; glycogen is a branched form, where the glucose
monomers are joined by alpha-glycosidic linkages.
Depending upon the molecules attached that are of the following types 1.
Glucan - A polysaccharide of glucose 2. Fructan - A polysaccharide
of fructose 3. Galactan - A polysaccharide of galactose 4. Araban - A
polysaccharide of arabinose 5. Xylan - A polysaccharide of xylose.

Homopolysaccharides (homoglycans) consist of a single type

of monomer. Cellulose and starch are the best-known examples. However,
they have very different properties. Cellulose consists of β-(1 → 4)-
linked glucose units arranged in a ribbon-type conformation in a zigzag
pattern.

Parallel chains fit closely to each other and associate with

multiple hydrogen bonds to give rise to long fibers, which are totally
insoluble in water and relatively inert. Although subject to some swelling in
water, cellulose is completely unaffected by boiling in water. Natural
cellulose (cellulose I) has considerable crystallinity (60–80%), as revealed
by X-ray diffraction. Starchconsists of two
components, viz. amylose (essentially linear) and amylopectin (branched).

The α-(1 → 4)-linked glucose molecules in amylose allow considerable

conformational freedom. In freshly prepared dilute solution, amylose is in
the form of a random coil. On standing at ambient temperature, it
‘retrogrades’, i.e., the random coils form double helical associations, which
are insoluble and sediment as a white precipitate.

At a high concentration, free settling is prevented, and gels are formed as

the randomly oriented chains approach each other at several junction
zones. Double helices are formed at the junctions, which do not extend the
full length of the molecule.

The net result is a three-dimensional structure, whose rigidity depends on

the concentration of amylose and hence the number of junction zones per
unit volume. Amylose gels are translucent to almost opaque, owing
to light scattering. Double helices of retrograded amylose are dissociated at
a temperature of about 160 °C.

Amylopectin consists of α-(1 → 4)-linked glucose units (95%) and α-

(1 → 6)-linked branched points (5%) and is fairly soluble in water. The outer
chains of amylopectin, consisting of about 15–25 glucose units, because of
their close proximity readily form short double helices during
starch biosynthesis and within food systems upon storage. Amylopectin is
fairly stable in solution and forms soft gels at a high concentration.
Waxy maize starch consists almost entirely of amylopectin and gives clear
or slightly translucent weak gels.

The term starch refers to a group of plant reserve polysaccharides

consisting almost exclusively of a linear component (amylose) and a
branched component (amylopectin).

The use of starch as an energy source by humans depends on the

ability to convert it completely to individual glucose units; the process is
initiated by the action of enzymes called amylases, synthesized by
the salivary glands in the mouth, and continues in the intestinal tract.

The primary product of amylase action is maltose, which is hydrolyzed

to two component glucose units as it is absorbed through the walls of
the intestine.
Blood Buffer?
Buffer solutions are extremely important in biology and medicine because
most biological reactions and enzymes need very specific pH ranges in
order to work properly.

Human blood contains a buffer of carbonic acid (H 2CO3) and bicarbonate

anion (HCO3-) in order to maintain blood pH between 7.35 and 7.45, as a
value higher than 7.8 or lower than 6.8 can lead to death. In this buffer,
hydronium and bicarbonate anion are in equilibrium with carbonic acid.

Furthermore, the carbonic acid in the first equilibrium can decompose into
CO2 gas and water, resulting in a second equilibrium system between
carbonic acid and water. Because CO2 is an important component of the
blood buffer, its regulation in the body, as well as that of O 2 , is extremely
important. The effect of this can be important when the human body is
subjected to strenuous conditions.
In the body, there exists another equilibrium between hydronium and
oxygen which involves the binding ability of hemoglobin. An increase in
hydronium causes this equilibrium to shift towards the oxygen side, thus
releasing oxygen from hemoglobin molecules into the surrounding
tissues/cells. This system continues during exercise, providing continuous
oxygen to working tissues.
In summation, the blood buffer is:
H3O++HCO−3⇌H2CO3+H2O(1)(1)H3O++HCO3−⇌H2CO3+H2O
With the following simultaneous equilibrium:
H2CO3⇌H2O+CO2(2)
Bicarbonate buffer (HCO3–/CO2)
Bicarbonate buffer is the most important buffer system in blood
plasma (generally in the extracellular fluid). This buffer consists of weak
acid H2CO3 (pK1 = 6,1) and conjugated base HCO3– (bicarbonate).
Bicarbonate concentration is given in mmol/l (average value is 24 mmol/l).
Since carbonic acid is very unstable molecule measurement of its
concentration is very difficult. H2CO3 is produced from CO2 hence it is
possible to express carbonic acid concentration as partial pressure of
CO2 (pCO2) because pCO2 is directly proportional to CO 2 concentration.
pCO2 is easily measured (kPa, mmHg). Average value in arterial blood
is 5,3 kPa = 40 mmHg. pCO2multiplied by α gives us molar concentration
of dissolved CO2 (α = 0,226 for pCO2 in kPa, α = 0,03 if pCO2 for mmHg).
Conversion relationship between mmHg and kPa is: 1 Pa = 0,0075 mmHg
(i.e. 760 mmHg ≈ 100 kPa). In normal plasma pH is HCO 3–/CO2 ratio 20 / 1.

Protein buffers
Body proteins (plasma proteins and intracellular) are the
most abundant and the most powerful buffer system in whole
organism. Some amino acids have acid or basic side chains (His, Lys, Arg,
Glu, Asp). Among blood proteins haemoglobin is the most important. It
provides 35 % of buffering capacity of blood, remaining proteins provide
only 7 %.

Phosphate buffer
Phosphate buffer consists of inorganic and organic bound phosphate
(i.e. esters of organic substances, e.g. AMP, ADP, and ATP). Phosphate
buffer is important intracellular and urine buffer. In blood it accounts for
only 5 % of buffering capacity.

PHOSPHOLIPIDS?
Phospholipids are a class of lipids that are a major component of all cell
membranes. They can form lipid bilayers because of
their amphiphilic characteristic. The structure of the phospholipid molecule
generally consists of two hydrophobic fatty acid "tails" and a hydrophilic"
head" consisting of a phosphate group.
The two components are joined together by a glycerol molecule. The
phosphate groups can be modified with simple organic molecules such
as choline, ethanolamine or serine.
The first phospholipid identified in 1847 as such in biological tissues
was lecithin, or phosphatidylcholine, in the egg yolk of chickens by the
French chemist and pharmacist, Theodore Nicolas Gobley. Biological
membranes in eukaryotes also contain another class of lipid, sterol,
interspersed among the phospholipids and together they provide
membrane fluidity and mechanical strength.
Purified phospholipids are produced commercially and have found
applications in nanotechnology and materials science.
An amphiphile (from the Greek αμφις, amphis: both and φιλíα, philia: love,
friendship; also termed "amphipathic") is a term describing a chemical
compound possessing both hydrophilic (water-loving, polar) and lipophilic
(fat-loving, non-polar) properties. The phospholipid head contains a
negatively charged phosphate group and glycerol; it is hydrophilic.

The phospholipid tails usually consist of 2 long fatty acid chains; they are
hydrophobic and avoid interactions with water. When placed in aqueous
solutions, phospholipids are driven by hydrophobic interactions that result
in the fatty acid tails aggregating to minimize interactions with water
molecules.
These specific properties allow phospholipids to play an important role in
the phospholipid bilayer. In biological systems, the phospholipids often
occur with other molecules (e.g., proteins, glycolipids, sterols) in
a bilayer such as a cell membrane. Lipid bilayers occur when hydrophobic
tails line up against one another, forming a membrane of hydrophilic heads
on both sides facing the water.
Such movement can be described by the fluid mosaic model, that
describes the membrane as a mosaic of lipid molecules that act as a
solvent for all the substances and proteins within it, so proteins and lipid
molecules are then free to diffuse laterally through the lipid matrix and
migrate over the membrane.
Sterols contribute to membrane fluidity by hindering the packing together of
phospholipids. However, this model has now been superseded, as through
the study of lipid polymorphism it is now known that the behaviour of lipids
under physiological (and other) conditions is not simple
APPLICATIONS OF PHOSPHOLIPIDS
Phospholipids have been widely used to prepare liposomal, ethosomal and
other nanoformulations of topical, oral and parenteral drugs for differing
reasons like improved bio-availability, reduced toxicity and increased
permeability across membranes. Liposomes are often composed
of phosphatidylcholine-enriched phospholipids and may also contain mixed
phospholipid chains with surfactant properties. The ethosomal formulation
of ketoconazole using phospholipids is a promising option for transdermal
delivery in fungal infections.

Creatinine clearance test?

Creatinine is a waste product from the normal breakdown of muscle tissue. As
creatinine is produced, it's filtered through the kidneys and excreted in urine.
Doctors measure the blood creatinine level as a test of kidney function.
The kidneys' ability to handle creatinine is called the creatinine clearance rate,
which helps to estimate the glomerular filtration rate (GFR) -- the rate
of blood flow through the kidneys.

Creatinine and Creatinine Clearance Test

Creatinine is a waste product that is produced continuously during normal

muscle breakdown. The kidneys filter creatinine from the blood into the urine,
and reabsorb almost none of it.

The amount of blood the kidneys can make creatinine-free each minute is
called the creatinine clearance. Creatinine clearance in a healthy young
person is about 95 milliliters per minute for women/120 milliliters per minute
for men. This means that each minute, that person's kidneys clear 95-120 mL
of blood free of creatinine. The GFR can vary depending on age, sex, and
size. Generally, the creatinine clearance is a good estimation of the
glomerular filtration rate.
Creatinine and creatinine clearance tests to check renal function (kidney
function). Testing the rate of creatinine clearance shows the kidneys' ability to
filter the blood. As renal function declines, creatinine clearance also goes
down.
There are two main ways to use creatinine tests to measure kidney function:

 Creatinine clearance can be precisely determined by measuring the

amount of creatinine present in a sample of urine collected over 24
hours. This method requires a person to place all his urine in a plastic
jug for one day, then bring it in for testing. Although the urine creatinine
measurement method is inconvenient, it may be necessary to diagnose
some kidney conditions.

 GFR can be estimated using a single blood level of creatinine, which

your doctor enters into a formula. Different formulas are available, which
take into account age, sex, and sometimes weight and ethnicity. The
higher the blood creatinine level, the lower the estimated GFR and
creatinine clearance.

Vitamin A deficiency?
Vitamin A deficiency (VAD) or hypovitaminosis A is a lack of vitamin
A in blood and tissues. It is common in poorer countries, but rarely is seen
in more developed countries. Nyctalopia (night blindness) is one of the first
signs of VAD. Xerophthalmia, keratomalacia, and complete blindness can
also occur since vitamin A has a major role in phototransduction. The three
forms of vitamin A include retinols, beta-carotenes, and carotenoids.
Vitamin A deficiency is the leading cause of preventable childhood
blindness, and is critical to achieving Millennium Development Goal 4 to
reduce child mortality. About 250,000 to 500,000 malnourished children in
the developing world go blind each year from a deficiency of vitamin A,
around half of whom die within a year of becoming blind.
The prevalence of night blindness due to VAD is also high among pregnant
women in many developing countries. VAD also contributes to maternal
mortality and other poor outcomes in pregnancy and lactation.
VAD also diminishes the ability to fight infections. In countries where
children are not immunized, infectious diseases such as measles have
higher fatality rates.
As elucidated by Alfred Sommer, even mild, subclinical deficiency can also
be a problem, as it may increase children's risk of developing respiratory
and diarrheal infections, decrease growth rate, slow bone development,
and decrease likelihood of survival from serious illness.
VAD is estimated to affect about one-third of children under the age of five
around the world. It is estimated to claim the lives of 670,000 children
under five annually.
Around 250,000–500,000 children in developing countries become blind
each year owing to VAD, with the highest prevalence in Southeast Asia and
Africa. According to the World Health Organization (WHO), VAD is under
control in the United States, but in developing countries, VAD is a
significant concern. Globally, 65% of all children aged 6 to 59 months
received two doses of vitamin A in 2013, fully protecting them against VAD
(80% in the least developed countries).
Sign and symptoms

Visual loss, Bitot’s spot, Imtiaz’s sign, impaired immune function.

Night Blindness

Night blindness is the difficulty for the eyes to adjust to dim light. Affected
individuals are unable to distinguish images in low levels of illumination.
People with night blindness have poor vision in the darkness, but see
normally when adequate light is present.
VAD affects vision by inhibiting the production of rhodopsin, the eye
pigment responsible for sensing low-light situations. Rhodopsin is found in
the retina and is composed of retinal (an active form of vitamin A)
and opsin (a protein). Because the body cannot create retinal in sufficient
amounts, a diet low in vitamin A leads to a decreased amount of rhodopsin
in the eye, as the retinal is inadequate to bind with opsin. Night blindness
results.
Night blindness caused by VAD has been associated with the loss of goblet
cells in the conjunctiva, a membrane covering the outer surface of the eye.
Goblet cells are responsible for secretion of mucus, and their absence
results in xerophthalmia, a condition where the eyes fail to produce tears.
Dead epithelial and microbial cells accumulate on the conjunctiva and form
debris that can lead to infection and possibly blindness.
Decreasing night blindness requires the improvement of vitamin A status in
at-risk populations. Supplements and fortification of food have been shown
to be effective interventions. Supplement treatment for night blindness
includes massive doses of vitamin A (200,000 IU) in the form of retinyl
palmitate to be taken by mouth, which is administered two to four times a
year. Intramuscular injections are poorly absorbed and are ineffective in
delivering sufficient bioavailable vitamin A. Fortification of food with vitamin
A is costly, but can be done in wheat, sugar, and milk. Households may
circumvent expensive fortified food by altering dietary habits. Consumption
of yellow-orange fruits and vegetables rich in carotenoids, specifically beta-
carotene, provides provitamin A precursors that can prevent VAD-related
night blindness. However, the conversion of carotene to retinol varies from
person to person and bioavailability of carotene in food varies.
Infection Rate

Along with poor diet, a large amount of infection and disease is present in
many developing communities. Infection is very draining on vitamin A
reserves and this vitamin A deficit leaves the individual more susceptible to
infection; increased documentation of xerophthalmia has been seen after
an outbreak of measles and the varying stages of xerophthalmia become a
good reference point for the extent of deficiency (with mortality increasing
with severity of the eye disease). In a longitudinal study of preschool
Indonesian children, susceptibility to disease increased nine times when
severe VAD was present.
The reason for the increased infection rate in vitamin A deficient
populations is the T-killer cells require the retinol metabolite retinoic acid to
proliferate correctly. Retinoic acid is a ligand for nuclear retinoic acid
receptors that bind the promoter regions of specific genes, thus activating
transcription and stimulating T cell replication. A vitamin A-deficient diet will
have a very limited amount of retinol, so cell proliferation and replication will
be suppressed, contributing to a reduced number of T-cells and
lymphocytes. Suppression of these result in a lack of immune reaction if
pathogens become present in the body and consequently a greater
susceptibility to incubation of disease.
VAD and infections aggravate each other, so with infection, vitamin A levels
are depleted, which in turn reduces intestinal absorption of vitamin A. Very
often seen with VAD is protein energy malnutrition, in which the synthesis
of retinol binding protein (RBP) is decreased, consequently the uptake of
retinol is reduced. This leads to an inability to use any vitamin A present as
the RBP is absent, so the retinol cannot be transported to the liver,
maximising the VAD.

Functions of Plasma Proteins?

Serum or plasma proteins are primarily synthesized in the liver; a
smaller percentage due to immunoglobulins is produced by
lymphocytes and plasma cells. Total protein consists of albumin,
globulins, and fibrinogen (in plasma only). Proteins function to
control oncotic pressure, transport substances (hemoglobin, lipids,
calcium), and promote inflammation and the complement cascade.
Changes in total protein levels are due mostly to changes in albumin
concentration.

Plasma proteins play a limited role in extracellular buffering, whereas

intracellular proteins play an important role in the total buffer
response of the body. The buffer effect of proteins is the result of their
dissociable side groups. For most proteins, including hemoglobin, the
most important of these dissociable groups is the imidazole ring
of histidineresidues (pKa, 6.4 to 7.0). Amino-terminal amino groups
(pKa, 7.4 to 7.9) also contribute somewhat to the buffer effect of
proteins.

Albumin and Serum Protein Markers

Serum protein markers, most notably albumin, have demonstrated

significant value in assessing nutritional status and predicting outcomes in
the setting of patients considered for elective surgery. Unfortunately the
value of serum protein levels as indicators of nutritional status is extremely
limited during the acute-phase response to injury, inflammation, infection,
and surgical stress. The physiologic stress response upregulates
expression of acute-phase reactive proteins, with unpredictable effects on
“marker” protein levels. In the setting of acute burn care, presenting
albumin levels or short-term changes therein should not be interpreted as
being indicative of nutritional progress.

Protein Markers for Proteinuria and Hematuria

Excess urinary protein loss can result from increased permeability of the
glomeruli to the passage of serum proteins (glomerular proteinuria),
decreased reabsorption of proteins by the renal tubules(tubular
proteinuria), or increased secretion of tissue protein into the urine
(secretory proteinuria).

Additionally, increased excretion of low molecular weight proteins may be

due to marked overproduction of the protein, resulting in the filtered load
exceeding the normal proximal reabsorptive capacity (overflow proteinuria).

Serum Enzymes?
Certain tissue cells contain characteristic enzymes which enter the blood
only when the cells to which they are confined are damaged or destroyed.
The presence in the blood of significant quantities of these specific
enzymes indicates the probable site of tissue damage

Aldolase:

Aldolase is present most significantly in skeletal and heart muscle. Damage

to skeletal muscle produces high serum levels of aldolase, particularly in
the case of progressive muscular dystrophy. Aldolase may also be slightly
increased in early stages of viral hepatitis and advanced cancer of the
prostate.
Creatine Phosphokinase (CPK or CK):

CPK catalyzes the reversible transfer of phosphate groups between

creatine and phosphocreatine as well as between ATP and ADP. Most of
the CPK resides in skeletal muscle, heart muscle, and in the
gastrointestinal tract. CPK enters the blood rapidly following damage to
muscle cells. At first CPK seemed to be an excellent marker for acute
myocardial infarction (heart damage) or skeletal muscle damage.
Unfortunately, the CPK levels rise and fall rapidly and coincide with a
variety of other circumstances including surgical procedures, vigorous
exercise, a fall, or a deep intramuscular injection. The measurement of
CPK levels still provides valuable differentiating diagnostic information.

Gamma-glutamyl Transpeptidase (GGT):

GGT catalyzes the transfer of the glutamyl groups among different

polypeptides and amino acids. Clinically significant GGT found in the blood
comes from cells that line the biliary tract. GGT levels rise dramatically with
obstructive diseases of the biliary tract and liver cancers. GGT is especially
useful in assessing liver function associated with alcohol - induced liver
disease.

Lactic Dehydrogenase (LDH):

This enzyme catalyzes the reversible reaction between pyruvic and lactic
acids.

LDH is present in nearly all types of metabolizing cells, but different cells
have different forms of the enzyme which can be distinguished. The
enzyme is especially concentrated in the heart, liver, red blood cells,
kidneys, muscles, brain, and lungs.

The total LDH can be further separated into five components or fractions
labeled by number: LDH-1, LDH-2, LDH-3, LDH-4, and LDH-5. Each of
these fractions, called isoenzymes, is used mainly by a different set of cells
or tissues in the body. The LDH isoenzymes test assists in differentiating
heart attack, anemia, lung injury, or liver disease from other conditions that
may cause the same symptoms.
LDH-1 is found mainly in the heart. LDH-2 is primarily associated with the
system in the body that defends against infection. LDH-3 is found in the
lungs and other tissues, LDH-4 in the kidney, placenta, and pancreas, and
LDH-5 in liver and skeletal muscle. Normally, levels of LDH-2 are higher
than those of the other isoenzymes.

Certain diseases have classic patterns of elevated LDH isoenzyme levels.

For example, an LDH-1 level higher than that of LDH-2 is indicative of a
heart attack or injury; elevations of LDH-2 and LDH-3 indicate lung injury or
disease; elevations of LDH-4 and LDH-5 indicate liver or muscle disease or
both. A rise of all LDH isoenzymes at the same time is diagnostic of injury
to multiple organs.

One of the most important diagnostic uses for the LDH isoenzymes test is
in the differential diagnosis of myocardial infarction or heart attack. The
total LDH level rises within 24-48 hours after a heart attack, peaks in two to
three days, and returns to normal in approximately five to ten days.

This pattern is a useful tool for a delayed diagnosis of heart attack. The
LDH-1 isoenzyme level, however, is more sensitive and specific than the
total LDH. Normally, the level of LDH-2 is higher than the level of LDH-1.
An LDH-1 level higher than that of LDH-2, a phenomenon known as
"flipped LDH," is strongly indicative of a heart attack.

Lipase:

Lipase is an enzyme secreted by the pancreas into the duodenum.

Damage to the pancreas as in acute pancreatitis results in lipase in the
blood from the secretory cells.

Transaminases (GOT and GPT):

Glutamic-Oxaloacetic Transaminase (GOT) occurs in large concentrations

in the heart and liver with moderate amounts in skeletal muscle, kidneys,
and pancreas. GOT levels can be used to diagnose myocardial infarction
within 10-48 hours. Other conditions with elevated GOT include
arrhythmias and severe angina of the heart, and liver damage.

Glutamic-Pyruvic Transaminase (GPT) is found in significant quantities in

liver, kidney, and skeletal muscle, in decreasing order. When liver cells are
damaged, GOT and GPT levels rise especially early in the disease. In
hepatitis, transaminase levels rise several days before jaundice begins.
The enzyme levels are especially useful in assessing subtle and early
changes in biliary obstruction and active cirrhosis.

PENTOSE PHOSPHATE PATHWAY or

Hexose Monophosphate Shunt?

The breakdown of the simple sugar, glucose, in glycolysis provides the first
6-carbon molecule required for the pentose phosphate pathway. During the
first step of glycolysis, glucose is transformed by the addition of a
phosphate group, generating glucose-6-phosphate, another 6-carbon
molecule. The pentose phosphate pathway can use any available
molecules of glucose-6-phosphate, whether they are produced by
glycolysis or other methods.

Two phases of the pentose phosphate pathway: 1) The oxidative phase

and 2) The non-oxidative phase.

The oxidative phase:

The “oxidative” word of this phase comes from the process of oxidation.
Oxidation is the breakdown of a molecule as it loses at least one of its
electrons. This phase is made up of 2 irreversible steps:

Step 1:

Glucose-6-phosphate is oxidized to form lactone. NADPH is produced as a

byproduct of this reaction as NADP+ is reduced as glucose-6-phosphate is
oxidized. Following the oxidation of glucose-6-phosphate, another reaction,
catalyzed by a different enzyme, uses water to form 6-phosphogluconate,
the linear product.

NADPH is similar in structure and function as the high energy electron

shuttle, NADH, mentioned in the cellular respiration articles. NADPH has
an added phosphate group and is used in the cell to donate its electrons,
just like NADH. Once NADPH has donated its electrons it is said to be
oxidized (oxidation = loss of electrons) and is now symbolized as, NADP +.
NADPH is often used in reactions that build molecules and occurs in a high
concentration in the cell, so that it is readily available for these types of
reactions.

Step 2:

Next, a carbon is removed (cleaved) and CO2 is released. Once again, the
electrons released from this cleavage is used to reduce NADP + to NADPH.
This new 5-carbon molecule is called ribulose-5-phosphate.

The non-oxidative phase:

The non-oxidative phase is really handy because these reactions

are reversible. This allows different molecules to enter the pentose
phosphate pathway in different areas of the non-oxidative phase and be
transformed up until the first molecule of the non-oxidative phase (ribulose-
5-phosphate). Ribulose-5-phosphate is the precursor to the sugar that
makes up DNA and RNA, and is also a product of the oxidative stage.

Step 3:
Ribulose-5-phosphate can be converted into two different 5-carbon
molecules. One is the sugar used to make up DNA and RNA called, ribose-
5-phosphate and this is the molecule we will focus on. Ribulose-5-
phosphate isn’t being divided because the carbon count is the same in the
next step.

Step 4:

The rest of the cycle is now made up of different options that depend on the
cell’s needs. The ribose-5-phosphate from step 3 is combined with another
molecule of ribose-5-phosphate to make one, 10-carbon molecule. Excess
ribose-5-phosphate, which may not be needed for nucleotide biosynthesis,
is converted into other sugars that can be used by the cell for metabolism.

The 10-carbon molecule is interconverted to create a 3-carbon molecule

and a 7-carbon molecule. The 3-carbon product can be shipped over to
glycolysis if it needs. That being said, recall that we can also work our way
back up to another molecule in this phase. So that 3-carbon molecule could
also be shipped over from glycolysis and transformed into ribose-5-
phosphate for DNA and RNA production.

Step 5:

The 3-carbon molecule and the 7-carbon molecule, from the

interconversion above in step 4, interconvert again to make a new 4-carbon
molecule and 6-carbon molecule. The 4-carbon molecule is a precursor for
amino acids, while the 6-carbon molecule can be used in glycolysis. The
same reversal of steps in option 4 can happen here as well.
The pentose phosphate pathway takes place in the cytosol of the cell, the
same location as glycolysis. The two most important products from this
process are the ribose-5-phosphate sugar used to make DNA and RNA,
and the NADPH molecules which help with building other molecules.
Diagram of the pentose phosphate pathway showing substrates and
enzymes involved in it. Tkt, transketolase; Gpi1, glucose phosphate
isomerase 1; G6pd2, glucose-6-phosphate dehydrogenase 2; Rpe,
ribulose-5-phosphate-3-epimerase; Rpia, ribose 5-phosphate isomerase A;
Taldo1, transaldoase 1; Rbks, ribokinase; Prps1, phosphoribosyl
pyrophosphate synthetase 1.

Basal Metabolic Rate (BMR)?

Even when resting, your body burns calories by performing basic functions
to sustain life, such as:

 breathing
  circulation

 nutrient processing

 cell production

Basal metabolic rate is the number of calories your body needs to

accomplish its most basic (basal) life-sustaining functions.

BASAL METABOLIC RATE versus RESTING METABOLIC RATE

Basal metabolic rate (BMR) is often used interchangeably with resting

metabolic rate (RMR). While BMR is a minimum number of calories
required for basic functions at rest, RMR — also called resting energy
expenditure (REE) — is the number of calories that your body burns while
it’s at rest.

Although BMR and RMR slightly differ from each other, your RMR should
be an accurate estimate of your BMR.

Estimate BMR

One popular way to estimate BMR is through the Harris-Benedict formula,

which takes into account weight, height, age, and gender.

Women:

BMR = 655 + (9.6 × weight in kg) + (1.8 × height in cm) – (4.7 × age in
years)

Men:

BMR = 66 + (13.7 × weight in kg) + (5 × height in cm) – (6.8 × age in years)

BMR can be used to help gain, lose, or maintain your weight. By knowing
how many calories you burn, you can know how many to consume. To put
it simply:

If you’ve estimated your BMR using the Harris-Benedict formula, your next
step is to include the number of calories you burn during daily activities
based on your lifestyle:

 Sedentary. If you get minimal or no exercise, multiply your BMR by

1.2.

 Lightly active. If you exercise lightly one to three days a week,

multiply your BMR by 1.375.

 Moderately active. If you exercise moderately three to five days a

week, multiply your BMR by 1.55.

 Very active. If you engage in hard exercise six to seven days a

week, multiply your BMR by 1.725.

 Extra active. If you engage in very hard exercise six to seven days a
week or have a physical job, multiply your BMR by 1.9.

The final number is approximately how many calories you need on a daily
basis to maintain your weight.

Glucose Tolerance Test (GTT)?

The oral glucose tolerance test (OGTT) was the gold standard for making
the diagnosis of type 2 diabetes. It is still commonly used
during pregnancy for diagnosing gestational diabetes. With an
oral glucose tolerance test, the person fasts overnight (at least 8 hours, but
not more than 16 hours).
The next morning, the fasting plasma glucose is tested. After this test, the
person receives a dose of oral glucose (the dose depends upon the length
of the test). There are several methods employed by obstetricians to do this
test, but the one described here is standard. Usually, the glucose is in a
sweet-tasting liquid that the person drinks. Blood samples are taken up to
four times at different time points after consumption of the sugar to
measure the blood glucose.

For the glucose tolerance test to give reliable results, the person must be in
good health (not have any other illnesses, not even a common cold). Also,
the person should be normally active (not lying down, for example, as an
inpatient in a hospital) and should not be taking medicines that could affect
the blood glucose.

In preparation for the oral glucose tolerance test, the person should eat and
drink as they normally would. The morning of the test, the person should
not smoke or consume caffeine.

Glucose Tolerance Test Measure

The classic oral glucose tolerance test measures blood glucose levels five
times over a period of three hours. Some physicians simply take a baseline
blood sample followed by a sample two hours after drinking the glucose
solution. In a person without diabetes, the glucose levels rise and then fall
quickly. In someone with diabetes, glucose levels rise higher than normal
and fail to come back down as fast.

People with glucose levels between normal and diabetic levels have so-
called impaired glucose tolerance (IGT). People with impaired glucose
tolerance do not have diabetes.
Each year, 5% to 10% of people whose test results show impaired glucose
tolerance actually develop diabetes. Weight loss and exercise may help
people with impaired glucose tolerance return their glucose levels to
normal. In addition, some physicians advocate the use of medications,
such as metformin (Glucophage), to help prevent/delay the onset of overt
diabetes. Studies have shown that impaired glucose tolerance itself may be
a risk factor for the development of heart disease, and whether impaired
glucose tolerance turns out to be an entity that deserves treatment itself is
something that physicians are currently debating.

Preparation for Glucose tolerance test

As mentioned previously, preparation for the oral glucose tolerance test

involves fasting overnight (from 8 to 16 hours) and participating normally in
activities of daily living. The individual should eat and drink as they normally
do prior to the test. The morning of the test, the person should not
consume caffeine or smoke.

Evaluation of Glucose Tolerance Test

Glucose tolerance tests may lead to one of the following diagnoses:

 Normal response: A person is said to have a normal response when

the two hour glucose level is less than 140 mg/dl, and all values
between 0 and 2 hours are less than 200 mg/dl.

 Impaired glucose tolerance (IGT): A person is said to have impaired

glucose tolerance when the fasting plasma glucose is less than 126
mg/dl and the two hour glucose level is between 140 and 199 mg/dl.
This is sometimes referred to as "prediabetes" because people with
IGT have a higher risk of developing diabetes.
  Diabetes: A person has diabetes when two diagnostic tests done on
different days show that the blood glucose level is high. This means
either the two hour levels is greater than 200 mg/dl or the fasting
glucose is noted as greater than 126 mg/dl. A
glycosylated hemoglobin(HbA1c) level of 6.5% or more also supports
a diagnosis of diabetes mellitus.

 Diabetes during pregnancy: A pregnant woman has diabetes if she

has a fasting plasma glucose of over 92 mg/dl, or a two hour glucose
level greater than 153 mg/dl.

Glucose Tolerance Test during Pregnancy

As mentioned previously, the glucose tolerance test is used for the

diagnosis of gestational diabetes (diabetes that develops during
pregnancy). It may be used if there are equivocal fasting or random blood
glucose results, or to screen for gestational diabetes in pregnant women
between 24 to 28 weeks of gestation who are not known to have diabetes.

The test may also be used in the postpartum period to detect diabetes in
women who had gestational diabetes during pregnancy. Women who had
gestational diabetes do not always develop diabetes later in life, but they
should undergo diabetes screening at least every three years throughout
their life.

Isoenzymes and its Significance?

Isoenzymes are enzymes that catalyze identical chemical reactions but are
composed of different aminoacid sequences. They are called as
isoenzymes. Isoenzymes are produced by different genes. They occur in
many tissues throughout the body and are important for different
developmental and metabolic processes.

Isoenzymes are useful biochemical markers and can be measured in the

blood stream to diagnose medical conditions. Isoenzymes can be
differentiated from one another using gel electrophoresis.

Some clinically important isoenzymes are

Creatine Kinase (CK, CPK)

Lactate Dehydrogenase (LDH)

It is formed by the association of five peptide chains of two different kinds

of monomers: M and H

The variants seen in humans are:

LDH1: M M M M (abundant in heart, brain erythrocytes; around 33% of

serum LDH)

LDH2: M M M H (abundant in heart, brain erythrocytes; around 45% of

serum LDH)

LDH3: M M H H (abundant in brain, kidneys, lung; around 18 % of serum

LDH)

LDH4: M H H H ((abundant in liver, skeletal muscle, kidney; around 3% of

serum LDH)

LDH5: H H H H ((abundant in liver, skeletal muscle, ileum; around 1 % of

serum LDH)
In myocardial infarction, Total LDH increases, and since heart muscle
contains more LDH1 than LDH2, LDH1 becomes greater than LDH2
between 12 and 24 hours, after the infarction, so the ratio LDH1/LDH2
becomes higher than 1 and will stay flipped for several days.

An increase of LDH 5 in serum is seen in different hepatic pathologies:

cirrhosis, hepatitis and others. An increase of LDH5 in heart diseases
usually indicates secondary congestive liver involvement.
Creatine Kinase :

Creatine Kinase (CK) aka Creatine phosphokinase (CPK) is a similar

example: three isoenzymes formed by combinations of different subunits:

CK1 (BB) is abundant in brain and smooth muscle (practically absent form
serum)
CK2 (MB) is abundant in cardiac muscle, some in skeletal muscle
(practically absent from serum)
CK3 (MM) is abundant in skeletal muscle and cardiac muscle (practically
100 % of serum CK)
The primary clinical use of CK studies is the diagnosis of Myocardial
Infarction,
(increased in the MB variant), but CK is also increased in different
conditions
as muscular diseases and traumas (MM and MB) and brain trauma and
brain surgery (BB).

CK2 appears in serum within 6 hours after the myocardial infarction and is
cleared after 24 to 48 hours. A persistence of CK2 in serum indicates
extension of the infarction to other areas or another infarction.

Functions of Vitamin C?
Vitamin C (L-ascorbic acid and its reduced form, dehydroascorbic acid) is a
water-soluble vitamin whose best-defined function is as a cofactor for the
enzyme required in the hydroxylation of proline and lysine in collagen
formation. It can be synthesized by many mammals, but not by humans.

The highest vitamin C content is found in green and red peppers, broccoli,
citrus fruits, strawberries, melons, tomatoes, raw cabbage, potatoes, and
leafy greens such as spinach, turnip, and mustard greens. Meat, fish,
poultry, eggs, and dairy products contain much smaller amounts, and
cereal grains contain essentially none. Losses of vitamin C occur when
foods are cooked in large amounts of water, exposed to extensive heating,
or exposed to air.

Vitamin C has other functions in addition to its role in collagen synthesis.

Along with its role in hydroxylation reactions, vitamin C affects leukocyte
and macrophage function, immune response, wound healing, and allergic
reactions. The involvement of vitamin C in these areas is less well
documented, and the levels necessary to achieve these benefits are not
known but are assumed to be much higher (pharmacological) than those
required for scurvy prevention.

Thus, the priority for adding vitamin C based on these roles cannot be
established. However, if cost-effectiveness analyses show that providing
higher fortification levels to prevent scurvy is not warranted, it is extremely
unlikely that better knowledge about the other possible benefits of vitamin
C would result in favorable cost-effective analysis for this objective.
Vitamin C in the diet can enhance the absorption of iron from plant sources
(non-heme iron) and improve the absorbability of fortification iron
(nonchelated inorganic iron) added to diets that contain inhibitors of iron
absorption (e.g., the phytate and polyphenols found in CSB and WSB).

The effects of ascorbic acid and of foods containing the same amount of
ascorbic acid appear to be the same. A two to threefold increase in
absorption of non-heme food iron from a meal can be expected from
adding foods that contain about 50–100 mg of ascorbic acid. In the past
few years, concern about the prevalence of iron deficiency anemia in
recipient populations (which is undoubtedly vastly higher than the
prevalence of ascorbic acid deficiency) has provoked recommendations to
increase the present level of iron in commodities from 15 to 30 mg/kg.

However, this may result in organoleptic problems, as well as accelerated

ascorbic acid oxidation during storage. It is the committee's understanding
that the feasibility of increasing the ferrous fumarate content of WSB and
CSB is as yet unresolved. However, the efficiency of absorption of ferrous
iron added to CSB is likely to be relatively poor. The addition of about 25
mg of ascorbic acid to a meal approximately doubles the percentage of
non-heme iron absorbed. However, data on the effect of increased levels of
vitamin C added to corn-soy milk (CSM) showed no significant
improvement in iron absorption.
Types of Immunoglobulins?
IgG
IgG is the most common class of immunoglobulin. It is present in the
largest amounts in blood and tissue fluids. Each IgG molecule consists of
the basic four-chain immunoglobulin structure—two identical H chains and
two identical L chains (either kappa or lambda)—and thus carries two
identical antigen-binding sites.
There are four subclasses of IgG, each with minor differences in its H
chains but with distinct biological properties. IgG is the only class of
immunoglobulin capable of crossing the placenta; consequently, it provides
some degree of immune protection to the developing fetus. These
molecules also are secreted into the mother’s milk and, once they have
been ingested by an infant, can be transported into the blood, where they
confer immunity.

IgM

Serum IgM exists as a pentamer in mammals and comprises approximately

10% of normal human serum Ig content. It predominates in primary
immune responses to most antigens and is the most efficient complement-
fixing immunoglobulin. IgM is also expressed on the plasma membrane of
B lymphocytes as a monomer. In this form, it is a B cell antigen receptor,
with the H chains each containing an additional hydrophobic domain for
anchoring in the membrane. Monomers of serum IgM are bound together
by disulfide bonds and a joining (J) chain.

Each of the five monomers within the pentamer structure is composed of

two light chains (either kappa or lambda) and two heavy chains. Unlike in
IgG (and the generalized structure shown above), the heavy chain in IgM
monomers is composed of one variable and four constant regions, with the
additional constant domain replacing the hinge region. IgM can recognize
epitopes on invading microorganisms, leading to cell agglutination.
This antibody-antigen immune complex is then destroyed by complement
fixation or receptor-mediated endocytosis by macrophages. IgM is the first
immunoglobulin class to be synthesized by the neonate and plays a role in
the pathogenesis of some autoimmune diseases.

The role of IgM in the immune response

Immunoglobulin M is the third most common serum Ig and takes one of two
forms:

 a pentamer where all heavy chains are identical and all light chains
are identical

 a monomer (e.g., found on B lymphocytes as B cell receptors)

The large pentameric structure allows for building of bridges between

encountered epitopes on molecules that are too distant as to be connected
by smaller IgG antibodies.

IgM is the first antibody built during an immune response. It is responsible

for agglutination and cytolytic reactions since in theory, its pentameric
structure gives it 10 free antigen-binding sites as well as it possesses a
high avidity.

Due to conformational constraints among the 10 Fab portions, IgM only has
a valence of 5. Additionally, IgM is not as versatile as IgG. However, it is of
vital importance in complement activation and agglutination.

IgM is predominantly found in the lymph fluid and blood and is a very
effective neutralizing agent in the early stages of disease. Elevated levels
can be a sign of recent infection or exposure to antigen.

IgA

IgA exists in serum in both monomeric and dimeric forms, comprising

approximately 15% of the total serum Ig. Secretory IgA, a dimer, provides
the primary defense mechanism against some local infections because of
its abundance in mucosal secretions (e.g., saliva and tears). The principal
function of secretory IgA may be not to destroy antigens but to prevent
passage of foreign substances into the circulatory system.
Properties of IgA:

 Molecular weight: 320,000 (secretory)

 H-chain type (MW): alpha (55,000)

 Serum concentration: 1 to 4 mg/mL

 Percent of total immunoglobulin: 15%

 Glycosylation (by weight): 10%

 Distribution: intravascular and secretions

 Function: protect mucus membranes

The role of IgA in the immune response

IgA comprises approximately 15% of all immunoglobulins in healthy serum.
Two IgA subtypes exist in humans, IgA1 und IgA2, while mice have only
one subclass. They differ in the molecular mass of the heavy chains and in
their concentration in serum.

IgA in serum is mainly monomeric, but in secretions, such as saliva, tears,

colostrums, mucus, sweat, and gastric fluid, IgA is found as a dimer
connected by a joining peptide. Most IgA is present in secreted form. This
is believed to be due to its properties in preventing invading pathogens by
attaching and penetrating epithelial surfaces. IgA is a very weak
complement-activating antibody; hence, it does not induce bacterial cell
lysis via the complement system. However, secretory IgA works together
with lysozymes (also present in many secreted fluids), which can hydrolyze
carbohydrates in bacterial cell walls thereby enabling the immune system
to clear the infection. IgA is predominantly found on epithelial cell surfaces
where it acts as a neutralizing antibody.

IgA deficiency in disease

The most prevalent antibody defect is a selective IgA deficiency (SIgAD).
Alterations in IgA1/IgA2 ratio often go hand in hand with specific disease
states, such as recurring infections of the airways or a kidney disorder
called IgA nephropathy.

There are various severe health conditions that can lead to low levels of
Immunoglobulin A in the body, one of which is gonorrhea. The bacteria that
cause gonorrhea produce an enzyme that splices IgA antibodies into Fc
and Fab fragments. The Fab part still can recognize the bacteria, but
without the Fc fragment, attachment to phagocyting cells is not possible.
When the body does not have sufficient quantities of IgA, the person may
be diagnosed with selective IgA deficiency.

Patients suffering from selective IgA deficiency can have normal levels of
the other antibodies, fully functioning T-cells, phagocytes, and other
components of the immune system. Patients who suffer from a selective
IgA deficiency are more prone to autoimmune disorders like rheumatoid
arthritis, lupus, allergies and asthma.

IgD

Properties of IgD:

 Molecular weight: 180,000

 H-chain type (MW): delta (70,000)

 Serum concentration: 0 to 0.4 mg/mL

 Percent of total immunoglobulin: 0.2%

 Glycosylation (by weight): 13%

 Distribution: lymphocyte surface

 Function: unknown

IgE

IgE and IgD are found in serum in much smaller quantities than other Igs.
IgE primarily defends against parasitic invasion and is responsible for
allergic reactions. Membrane IgD is a receptor for antigen found mostly on
mature B-lymphocytes.

Properties of IgE:

 Molecular weight: 200,000

 H-chain type (MW): epsilon (73,000)

 Serum concentration: 10 to 400 ng/mL

 Percent of total immunoglobulin: 0.002%

 Glycosylation (by weight): 12%

 Distribution: basophils and mast cells in saliva and nasal secretions

 Function: protect against parasites

Properties of IgD:

 Molecular weight: 180,000

 H-chain type (MW): delta (70,000)

 Serum concentration: 0 to 0.4 mg/mL

 Percent of total immunoglobulin: 0.2%

 Glycosylation (by weight): 13%

 Distribution: lymphocyte surface

 Function: unknown

The role of IgE in the allergic response

The heavy chain of IgE contains an extra domain, by which it attaches with
high affinity to Fc epsilon Receptor I (FcεRI) found primarily on eosinophils,
mast cells and basophils. When antigens such as pollen, venoms, fungus,
spores, dust mites or pet dander bind with the Fab portion of the IgE
attached to the cells, the cells degranulate and release factors like heparin,
histamine, proteolytic enzymes, leukotrienes and cytokines. As a
consequence, vasodilatation and increased small vessel permeability
causes fluid to escape from capillaries into the tissues, leading to the
characteristic symptoms of an allergic reaction. Most of these typical
allergic reactions like mucus secretion, sneezing, coughing or tear
production are considered beneficial to expel remaining allergens from the
body.

Studies have shown that conditions such as asthma, rhinitis, eczema,

urticaria, dermatitis and some parasitic infections (e.g., helminths and
tapeworms) lead to increased IgE levels. Binding of eosinophils with Fc
receptors to IgE-coated parasitic helminth worms results in death of the
parasite. Low levels of IgE can occur in a rare inherited disease that affects
muscle coordination (ataxia-telangiectasia).

Long Answers (10 Marks)

Glycolysis Pathway?
Glycolysis is the first step in the breakdown of glucose to extract energy for
cellular metabolism. Glycolysis consists of an energy-requiring phase
followed by an energy-releasing phase.

Glycolysis is a series of reactions that extract energy from glucose by

splitting it into two three-carbon molecules called pyruvates.

In organisms that perform cellular respiration, glycolysis is the first stage of

this process. However, glycolysis doesn’t require oxygen, and many
 anaerobic organisms—organisms that do not use oxygen—also have this
pathway. Glycolysis has ten steps.

Glycolysis takes place in the cytosol of a cell, and it can be broken down
into two main phases: the energy-requiring phase, and the energy-
releasing phase.

 Energy-requiring phase. In this phase, the starting molecule of

glucose gets rearranged, and two phosphate groups are attached to it. The
phosphate groups make the modified sugar—now called fructose-1,6-
bisphosphate—unstable, allowing it to split in half and form two phosphate-
bearing three-carbon sugars. Because the phosphates used in these steps
come from ATP, two ATP molecules get used up.
 The three-carbon sugars formed when the unstable sugar breaks
down are different from each other. Only one—glyceraldehyde-3-
phosphate—can enter the following step.

 Energy-releasing phase. In this phase, each three-carbon sugar is

converted into another three-carbon molecule, pyruvate, through a series of
reactions. In these reactions, two ATP molecules and one NADH molecule
are made. Because this phase takes place twice, once for each of the two
three-carbon sugars, it makes four ATP and two NADH overall.
 Each reaction in glycolysis is catalyzed by its own enzyme. The most
important enzyme for regulation of glycolysis
is phosphofructokinase, which catalyzes formation of the unstable,
two-phosphate sugar molecule, fructose-1,6-bisphosphate.
Phosphofructokinase speeds up or slows down glycolysis in response
to the energy needs of the cell.
  Overall, glycolysis converts one six-carbon molecule of glucose into
two three-carbon molecules of pyruvate. The net products of this
process are two molecules of ATP (4 ATP produced −2 ATP used up)
and two molecules of NADH.

Steps – Energy-requiring Phase

Step 1. A phosphate group is transferred from ATP to glucose, making

glucose-6-phosphate. Glucose-6-phosphate is more reactive than glucose,
and the addition of the phosphate also traps glucose inside the cell since
glucose with a phosphate can’t readily cross the membrane.

Step 2. Glucose-6-phosphate is converted into its isomer, fructose-6-

phosphate.

Step 3. A phosphate group is transferred from ATP to fructose-6-

phosphate, producing fructose-1,6-bisphosphate. This step is catalyzed by
the enzyme phosphofructokinase, which can be regulated to speed up or
slow down the glycolysis pathway.

Step 4. Fructose-1,6-bisphosphate splits to form two three-carbon sugars:

dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate.
They are isomers of each other, but only one—glyceraldehyde-3-phosphate
—can directly continue through the next steps of glycolysis.

Step 5. DHAP is converted into glyceraldehyde-3-phosphate. The two

molecules exist in equilibrium, but the equilibrium is “pulled” strongly
downward, in the scheme of the diagram above, as glyceraldehyde-3-
phosphate is used up. Thus, all of the DHAP is eventually converted.
Detailed steps: Energy-releasing phase

In the second half of glycolysis, the three-carbon sugars formed in the first
half of the process go through a series of additional transformations,
ultimately turning into pyruvate. In the process, four ATP molecules are
produced, along with two molecules of NADH.

Here, we’ll look in more detail at the reactions that lead to these products.
The reactions shown below happen twice for each glucose molecule since
a glucose splits into two three-carbon molecules, both of which will
eventually proceed through the pathway.

Step 6. Two half reactions occur simultaneously: 1) Glyceraldehyde-3-

phosphate (one of the three-carbon sugars formed in the initial phase) is
oxidized, and 2) NAD+ is reduced to NADH and H+. The overall reaction is
exergonic, releasing energy that is then used to phosphorylate the
molecule, forming 1,3-bisphosphoglycerate.

Step 7. 1,3-bisphosphoglycerate donates one of its phosphate groups

to ADP, making a molecule of ATP and turning into 3-phosphoglycerate in
the process.

Step 8. 3-phosphoglycerate is converted into its isomer, 2-

phosphoglycerate.

Step 9. 2-phosphoglycerate loses a molecule of water, becoming

phosphoenolpyruvate (PEP). PEP is an unstable molecule, poised to lose
its phosphate group in the final step of glycolysis.
Step 10. PEP readily donates its phosphate group to ADP, making a
second molecule of ATP. As it loses its phosphate, PEP is converted to
pyruvate, the end product of glycolysis.

KREB’S CYCLE or CITRIC ACID CYCLE or

TRICARBOXYLIC ACID CYCLE?
The pyruvate molecules generated during glycolysis are transported across
the mitochondrial membrane into the inner mitochondrial matrix, where they
are metabolized by enzymes in a pathway called the Krebs cycle.

The Krebs cycle is also commonly called the citric acid cycle or the
tricarboxylic acid (TCA) cycle. During the Krebs cycle, high-energy
molecules, including ATP, NADH, and FADH2, are created. NADH and
FADH2 then pass electrons through the electron transport chain in the
mitochondria to generate more ATP molecules.
The three-carbon pyruvate molecule generated during glycolysis moves
from the cytoplasm into the mitochondrial matrix, where it is converted by
the enzyme pyruvate dehydrogenase into a two-carbon acetyl coenzyme
A (acetyl CoA) molecule.

This reaction is an oxidative decarboxylation reaction. It converts the three-

carbon pyruvate into a two-carbon acetyl CoA molecule, releasing carbon
dioxide and transferring two electrons that combine with NAD+ to form
NADH. Acetyl CoA enters the Krebs cycle by combining with a four-carbon
molecule, oxaloacetate, to form the six-carbon molecule citrate, or citric
acid, at the same time releasing the coenzyme A molecule.

The six-carbon citrate molecule is systematically converted to a five-carbon

molecule and then a four-carbon molecule, ending with oxaloacetate, the
beginning of the cycle. Along the way, each citrate molecule will produce
one ATP, one FADH2, and three NADH. The FADH2 and NADH will enter
the oxidative phosphorylation system located in the inner mitochondrial
membrane. In addition, the Krebs cycle supplies the starting materials to
process and break down proteins and fats.

To start the Krebs cycle, citrate synthase combines acetyl CoA and
oxaloacetate to form a six-carbon citrate molecule; CoA is subsequently
released and can combine with another pyruvate molecule to begin the
cycle again. The aconitase enzyme converts citrate into isocitrate. In two
successive steps of oxidative decarboxylation, two molecules of CO2 and
two NADH molecules are produced when isocitrate dehydrogenase
converts isocitrate into the five-carbon α-ketoglutarate, which is then
catalyzed and converted into the four-carbon succinyl CoA by α-
ketoglutarate dehydrogenase.

The enzyme succinyl CoA dehydrogenase then converts succinyl CoA into
succinate and forms the high-energy molecule GTP, which transfers its
energy to ADP to produce ATP. Succinate dehydrogenase then converts
succinate into fumarate, forming a molecule of FADH2. Fumarase then
converts fumarate into malate, which malate dehydrogenase then converts
back into oxaloacetate while reducing NAD+ to NADH. Oxaloacetate is
then ready to combine with the next acetyl CoA to start the Krebs cycle
again. For each turn of the cycle, three NADH, one ATP (through GTP), and
one FADH2 are created. Each carbon of pyruvate is converted into CO2,
which is released as a byproduct of oxidative (aerobic) respiration.

Composition of cell. Structure and function of

cell Membrane?

There are many different types, sizes, and shapes of cells in the body. For
descriptive purposes, the concept of a "generalized cell" is introduced. It
includes features from all cell types. A cell consists of three parts: the cell
membrane, the nucleus, and, between the two, the cytoplasm. Within the
cytoplasm lie intricate arrangements of fine fibers and hundreds or even
thousands of miniscule but distinct structures called organelles.

Cell membrane
Every cell in the body is enclosed by a cell plasma membrane. The cell
membrane separates the material outside the cell, extracellular, from the
material inside the cell, intracellular. It maintains the integrity of a cell and
controls passage of materials into and out of the cell. All materials within a
cell must have access to the cell membrane (the cell's boundary) for the
needed exchange.

The cell membrane is a double layer of phospholipid molecules. proteins in

the cell membrane provide structural support, form channels for passage of
materials, act as receptor sites, function as carrier molecules, and provide
identification markers.

Nucleus and Nucleolus

The nucleus, formed by a nuclear membrane around a fluid nucleoplasm, is
the control center of the cell. Threads of chromatin in the nucleus contain
deoxyribonucleic acid (DNA), the genetic material of the cell. The nucleolus
is a dense region of ribonucleic acid (RNA) in the nucleus and is the site of
ribosome formation. The nucleus determines how the cell will function, as
well as the basic structure of that cell.

Cytoplasm
The cytoplasm is the gel-like fluid inside the cell. It is the medium for
chemical reaction. It provides a platform upon which other organelles can
operate within the cell. All of the functions for cell expansion, growth and
replication are carried out in the cytoplasm of a cell. Within the cytoplasm,
materials move by diffusion, a physical process that can work only for short
distances.

Cytoplasmic organelles
Cytoplasmic organelles are "little organs" that are suspended in the
cytoplasm of the cell. Each type of organelle has a definite structure and a
specific role in the function of the cell.

Structure of the plasma membrane

Each cell of your body is encased in a tiny bubble of membrane. The

plasma membrane not only defines the borders of the cell, but also allows
the cell to interact with its environment in a controlled way. Cells must be
able to exclude, take in, and excrete various substances, all in specific
amounts. In addition, they must able to communicate with other cells,
identifying themselves and sharing information.

To perform these roles, the plasma membrane needs lipids, which make a
semi-permeable barrier between the cell and its environment. It also needs
proteins, which are involved in cross-membrane transport and cell
communication, and carbohydrates (sugars and sugar chains), which
decorate both the proteins and lipids and help cells recognize each other.

Fluid mosaic model

The currently accepted model for the structure of the plasma membrane,
called the fluid mosaic model, was first proposed in 1972. This model has
evolved over time, but it still provides a good basic description of the
structure and behavior of membranes in many cells.

According to the fluid mosaic model, the plasma membrane is a mosaic of

components—primarily, phospholipids, cholesterol, and proteins—that
move freely and fluidly in the plane of the membrane. In other words, a
diagram of the membrane (like the one below) is just a snapshot of a
dynamic process in which phospholipids and proteins are continually sliding
past one another.
 The principal components of the plasma membrane are lipids
(phospholipids and cholesterol), proteins, and carbohydrate groups that are
attached to some of the lipids and proteins.

 A phospholipid is a lipid made of glycerol, two fatty acid tails, and a

phosphate-linked head group. Biological membranes usually involve two
layers of phospholipids with their tails pointing inward, an arrangement
called a phospholipid bilayer.

 Cholesterol, another lipid composed of four fused carbon rings, is

found alongside phospholipids in the core of the membrane.

 Membrane proteins may extend partway into the plasma membrane,

cross the membrane entirely, or be loosely attached to its inside or outside
face.

 Carbohydrate groups are present only on the outer surface of the

plasma membrane and are attached to proteins, forming glycoproteins, or
lipids, forming glycolipids.
The proportions of proteins, lipids, and carbohydrates in the plasma
membrane vary between different types of cells. For a typical human cell,
however, proteins account for about 50 percent of the composition by
mass, lipids (of all types) account for about 40 percent, and the remaining
10 percent comes from carbohydrates.

Phospholipids

Phospholipids, arranged in a bilayer, make up the basic fabric of the

plasma membrane. They are well-suited for this role because they
are amphipathic, meaning that they have both hydrophilic and
hydrophobic regions.

The hydrophilic, or “water-loving,” portion of a phospholipid is its head,

which contains a negatively charged phosphate group as well as an
additional small group which may also or be charged or polar. The
hydrophilic heads of phospholipids in a membrane bilayer face outward,
contacting the aqueous (watery) fluid both inside and outside the cell. Since
water is a polar molecule, it readily forms electrostatic (charge-based)
interactions with the phospholipid heads.

The hydrophobic, or “water-fearing,” part of a phospholipid consists of its

long, nonpolar fatty acid tails. The fatty acid tails can easily interact with
other nonpolar molecules, but they interact poorly with water. Because of
this, it’s more energetically favorable for the phospholipids to tuck their fatty
acid tails away in the interior of the membrane, where they are shielded
from the surrounding water. The phospholipid bilayer formed by these
interactions makes a good barrier between the interior and exterior of the
cell, because water and other polar or charged substances cannot easily
cross the hydrophobic core of the membrane.
In water or aqueous solution, phospholipids tend to arrange themselves
with their hydrophobic tails facing each other and their hydrophilic heads
facing out. If the phospholipids have small tails, they may form a micelle (a
small, single-layered sphere), while if they have bulkier tails, they may form
a liposome.

Proteins

Proteins are the second major component of plasma membranes. There

are two main categories of membrane proteins: integral and peripheral.

The portions of an integral membrane protein found inside the membrane

are hydrophobic, while those that are exposed to the cytoplasm or
extracellular fluid tend to be hydrophilic. Transmembrane proteins may
cross the membrane just once, or may have as many as twelve different
membrane-spanning sections. A typical membrane-spanning segment
consists of 20-25 hydrophobic amino acids arranged in an alpha helix,
although not all transmembrane proteins fit this model. Some integral
membrane proteins form a channel that allows ions or other small
molecules to pass.

Peripheral membrane proteins are found on the outside and inside

surfaces of membranes, attached either to integral proteins or to
phospholipids. Unlike integral membrane proteins, peripheral membrane
proteins do not stick into the hydrophobic core of the membrane, and they
tend to be more loosely attached.
 Carbohydrates

Carbohydrates are the third major component of plasma membranes. In

general, they are found on the outside surface of cells and are bound either
to proteins (forming glycoproteins) or to lipids (forming glycolipids).
These carbohydrate chains may consist of 2-60 monosaccharide units and
can be either straight or branched.

Along with membrane proteins, these carbohydrates form distinctive

cellular markers, sort of like molecular ID badges, that allow cells to
recognize each other. These markers are very important in the immune
system, allowing immune cells to differentiate between body cells, which
they shouldn’t attack, and foreign cells or tissues, which they should.

Membrane fluidity

The structure of the fatty acid tails of the phospholipids is important in

determining the properties of the membrane, and in particular, how fluid it
is.

Saturated fatty acids have no double bonds (are saturated with hydrogen),
so they are relatively straight. Unsaturated fatty acids, on the other hand,
contain one or more double bonds, often resulting in a bend or kink. The
saturated and unsaturated fatty acid tails of phospholipids behave
differently as temperature drops:

 At cooler temperatures, the straight tails of saturated fatty acids can

pack tightly together, making a dense and fairly rigid membrane.
 Phospholipids with unsaturated fatty acid tails cannot pack together
as tightly because of the bent structure of the tails. Because of this, a
membrane containing unsaturated phospholipids will stay fluid at lower
temperatures than a membrane made of saturated ones.

Most cell membranes contain a mixture of phospholipids, some with two

saturated (straight) tails and others with one saturated and one unsaturated
(bent) tail. Many organisms—fish are one example—can adjust
physiologically to cold environments by changing the proportion of
unsaturated fatty acids in their membranes.

Oxidation of fatty acid?

Triacylglycerols (fats) are the most abundant source of energy and
provide energy twice as much as carbohydrates and proteins. This is
achieved because the fatty acids which are present in the triacylglycerols
are already in the reduced form. To convert fats into energy, the digested
fats or the stored fats have to be first activated and transported to the
mitochondrial matrix as all the enzymes required for metabolism
(oxidation) are present there.

For activation of fatty acids, they are converted to fatty acyl-CoA by the
enzyme thiokinase in the cytosol and then with the help of carnitine (L-
carnitine is the active stereoisomer) is transported through the
mitochondrial membranes into the mitochondrial matrix.

Oxidation of fatty acids occurs in three stages:

 β-oxidation of fatty acids resulting in cleavage of two-carbon units (α

and β carbons) from the carboxyl end of fatty acyl-CoA with the
formation of acetyl CoA. This reaction keeps occuring till the entire
 fatty acyl chain is broken down to acetyl CoA molecules. For eg.
Palmitoyl CoA (16 carbon chain) on β-oxidation will give eight acetyl
CoA molecules. It is further discussed in detail below.
 The acetyl groups produced from β-oxidation of the fatty acid
participate in the Kreb’s cycle resulting in the formation of NADH and
FADH2.
 The reduced coenzymes (NADH and FADH2) are oxidized by giving
up the protons and electrons to oxygen present in the mitochondria to
synthesize ATP by oxidative phosphorylation in the Electron
Transport System.

Beta oxidation of Fatty Acids

Once the fatty acids have been transported to the mitochondrial matrix via
carnitine pathway, β-oxidation of fatty acyl-CoA (n carbons) occurs
within the mitochondria in four steps as discussed below:
 First step– Fatty acyl-CoA is acted upon by an enzyme acyl-CoA
dehydrogenase which is FAD dependent. Fatty acyl-
CoAundergoes dehydrogenation and forms a trans-double bond at
the α and β carbons to form trans-Δ2-enoyl-CoA. Acyl-CoA
dehydrogenase are present as three isoenzymes each specific for a
particular carbon chain length (short, intermediate and long). The
electrons which were removed from the fatty acyl-CoA chain are
transferred to FAD which gets reduced to FADH2. This
FADH2 immediately via the Electron Transport System gets converted
to ATP molecules.
 Second step– Enoyl-CoA hydratase catalyzes this reaction
where water is added. Hydration occurs at the double bond resulting
in the formation of β-hydroxyacyl-CoA.
 Third step– β-hydroxyacyl-CoA undergoes dehydrogenation to
form β-ketoacyl-CoA in the presence of β-hydroxyacyl-CoA
dehydrogenase. The electrons available as a result of
dehydrogenation are accepted by NAD+ to form NADH + H+ which
immediately exchanges these electrons with oxygen in the Electron
Transport System to form ATP molecules.
 Fourth step– This reaction is called as thiolysis as acyl-CoA
acetyltransferase(also known as thiolase) in the presence of CoA-
SH causes the cleavage of β-ketoacyl-CoA to form acetyl CoA and
the thioester of the original fatty acid with two carbons less. This
 cleavage occurs as the β carbon ketone group is a good target for
nucleophilic attack by the thiol (-SH) group of the coenzyme A.

The new fatty acyl-CoA (n-2 carbons) formed again participates in the β-
oxidation cycle to form a new fatty acyl-CoA with two carbons less (n-4
carbons) and a new molecule of acetyl CoA. This process continues till the
entire fatty acid is converted into acetyl CoA molecules.
Acetyl CoA formed from the above steps now enters the Kreb’s cycle to
get oxidized to CO2 and H2O.
Note:
 Oxidation of odd carbon length fatty acids has additional reactions
and is described in detail here: Oxidation of odd carbon chain
length fatty acids.
 For unsaturated fatty acids β-oxidation has two more additional steps
are required catalyzed by two enzymes isomerase and reductase.
Oxidation of unsaturated fats gives less energy as compared to
saturated fats. Read more about it here: Oxidation of unsaturated
fatty acids.
ACID – BASE BALANCE?
To maintain homeostasis, the human body employs many physiological
adaptations. One of these is maintaining an acid-base balance. In the
absence of pathological states, the pH of the human body ranges between
7.35 to 7.45, with the average at 7.40.
A pH at this level is ideal for many biological processes, one of the most
important being the oxygenation of blood. Also, many of the intermediates
of biochemical reactions in the body become ionized at a neutral pH, which
causes the utilization of these intermediates to be more difficult.
A pH below 7.35 is an acidemia, and a pH above 7.45 is an alkalemia. Due
to the importance of sustaining a pH level in the needed narrow range, the
human body contains compensatory mechanisms. This article intends to
impart a basic understanding of acid-base balance in the body while
providing a systematic way to approach patients who present with
conditions causing alterations in pH.
The human body experiences four main types of acid-based disorders:
metabolic acidosis, metabolic alkalosis, respiratory acidosis, and
respiratory alkalosis. If one of these conditions occurs, the human body
should induce a counterbalance in the form of an opposite condition. For
example, if a person is experiencing a metabolic acidemia, their body will
attempt to induce a respiratory alkalosis to compensate. It is rare for the
compensation to make the pH completely normal at 7.4. When using the
term acidemia or alkalemia, one is denoting that overall the pH is acidic or
alkalotic, respectively. While not necessary, it can be useful to employ this
terminology to distinguish between individual processes and the overall pH
status of the patient since multiple imbalances can happen at the same
time

Organ System Involved in pH Balance

Every organ system of the human body relies on pH balance; however, the
renal system and the pulmonary system are the two main modulators. The
pulmonary system adjusts pH using carbon dioxide; upon expiration,
carbon dioxide is projected into the environment. Due to carbon dioxide
forming carbonic acid in the body when combining with water, the amount
of carbon dioxide expired can cause pH to increase or decrease. When the
respiratory system is utilized to compensate for metabolic pH disturbances,
the effect occurs in minutes to hours.
The renal system affects pH by reabsorbing bicarbonate and excreting
fixed acids. Whether due to pathology or necessary compensation, the
kidney excretes or reabsorbs these substances which affect pH. The
nephron is the functional unit of the kidney. Blood vessels called glomeruli
transport substances found in the blood to the renal tubules so that some
can be filtered out while others are reabsorbed into the blood and recycled.
This is true for hydrogen ions and bicarbonate.
If bicarbonate is reabsorbed and/or acid is secreted into the urine, the pH
becomes more alkaline (increases). When bicarbonate is not reabsorbed or
acid is not excreted into the urine, pH becomes more acidic (decreases).
The metabolic compensation from the renal system takes longer to occur:
days rather than minutes or hours.

Function of pH Balance
The physiological pH of the human body is essential for many processes
necessary to life including oxygen delivery to tissues, correct protein
structure, and innumerable biochemical reactions that rely on the normal
pH to be in equilibrium and complete.
Oxygen Delivery to Tissues
The oxygen dissociation curve is a graph depicting the relationship of the
partial pressure of oxygen to the saturation of hemoglobin. This curve
relates to the ability of hemoglobin to deliver oxygen to tissues. If the curve
is shifted to the left, there is a decreased p50, meaning that the amount of
oxygen needed to saturate hemoglobin 50% is lessened and that there is
an increased affinity of hemoglobin for oxygen. A pH in the alkalotic range
induces this left shift. When there is a decrease in pH, the curve is shifted
to the right, denoting a decreased affinity of hemoglobin for oxygen.
Protein Structure
It would be hard to overstate the importance of proteins in the human body.
They makeup ion channels, carry necessary lipophilic substances
throughout our mostly lipophobic body, and participate in innumerable
biological processes. For proteins to complete necessary functions, they
must be in the proper configuration. The charges on proteins are what
allow their proper shape to exist. When pH is altered outside of the
physiological range, these charges are altered. The proteins are denatured
leading to detrimental changes in architecture that cause a loss of proper
function.
Biochemical Processes
Throughout the human body, many chemical reactions are in equilibrium.

When the levels of acid in your blood are too high, it’s called acidosis.
When your blood is too alkaline, it is called alkalosis.

Respiratory acidosis and alkalosis are due to a problem with the lungs.
Metabolic acidosis and alkalosis are due to a problem with the kidneys.

Each of these conditions is caused by an underlying disease or disorder.

Treatment depends on the cause.

Respiratory Acidosis

When you breathe, your lungs remove excess carbon dioxide from your
body. When they cannot do so, your blood and other fluids become too
acidic.

Symptoms of respiratory acidosis

Symptoms may include fatigue, shortness of breath, and confusion.

Causes of respiratory acidosis

There are several different causes of respiratory acidosis including:

 chest deformities or injuries

 chronic lung and airway diseases

 overuse of sedatives

 obesity

Types of respiratory acidosis

There are no noticeable symptoms of chronic respiratory acidosis. This is

due to the fact that your blood slowly becomes acidic and your kidneys
adjust to compensate, returning your blood to a normal pH balance.

Acute respiratory acidosis comes on suddenly, leaving the kidneys no time

to adjust. Those with chronic respiratory acidosis may experience acute
respiratory acidosis due to another illness that causes the condition to
worsen.

Diagnosis of respiratory acidosis

A complete physical examination is necessary. Diagnostic testing may

include:

 arterial blood gas test

 metabolic panel

 pulmonary function test

 chest X-ray

Treatment of respiratory acidosis

A doctor should be seen immediately to treat acute respiratory acidosis, as

this can be a life threatening condition. Treatment is targeted to the cause.
Bronchodilator medications may be given to correct some forms of airway
obstruction. If your blood oxygen level is too low, you may require oxygen.
Noninvasive positive pressure ventilation or a breathing machine may be
necessary.

To treat chronic respiratory acidosis, the underlying cause needs to be

determined in order for proper treatment to take place. The cause could be
from an organ deformity, an infection, or some type of inflammation. Each
cause may require a different treatment ranging from antibiotics to a
breathing machine.

Complications of respiratory acidosis

Respiratory acidosis is serious and requires immediate medical attention.

Potential complications of untreated respiratory acidosis include respiratory
failure, organ failure, and shock.

Preventing respiratory acidosis

You can take steps to help prevent some of the conditions that lead to
respiratory acidosis. Maintain a healthy weight. Take sedatives only under
strict doctor supervision and never combine them with alcohol. Do not
smoke.

METABOLIC ACIDOSIS

Metabolic acidosis occurs either when your body produces too much acid,
or when your kidneys are unable to remove it properly.

Symptoms of Metabolic acidosis

Symptoms can include rapid breathing, fatigue, and confusion.

Causes of Metabolic acidosis

There are three main types of metabolic acidosis. Diabetic acidosis,

or diabetic ketoacidosis, is a buildup of ketone bodies. This is usually due
to uncontrolled type 1 diabetes. Hyperchloremic acidosis is when your body
loses too much sodium bicarbonate, often after severe diarrhea.

Lactic acidosis is when too much lactic acid builds up. This can be due to:

 prolonged exercise

 lack of oxygen

 certain medications, including salicylates

 low blood sugar, or hypoglycemia

 alcohol

 seizures

 liver failure

 cancer

 kidney disease

 severe dehydration

 poisoning from consuming too much aspirin, ethylene glycol, and

methanol

Diagnosing metabolic acidosis

Diagnostic testing may include serum electrolytes, urine pH, and arterial
blood gases. Once acidosis is confirmed, other tests may be necessary to
pinpoint the cause.

Treatment of metabolic acidosis

The underlying condition behind the acidosis must be treated. In some

cases, sodium bicarbonate is prescribed to return the blood to a normal pH.

Complications of metabolic acidosis

Severe cases can lead to shock and can be life threatening.

ALKALOSIS

Alkalosis is when alkaline levels are too high due to decreased carbon
dioxide or increased bicarbonate. There are five kinds of alkalosis.

Symptoms of alkalosis

Symptoms of alkalosis may include:

 muscle twitching, hand tremor, muscle spasms

 numbness and tingling

 nausea

 vomiting

 lightheadedness
 confusion

Causes and types of alkalosis

Respiratory alkalosis is when your blood has low levels of carbon dioxide.
This can be caused by a number of factors, including:

 lack of oxygen

 high altitude

 fever

 lung disease

 liver disease

 salicylate poisoning

When you have alkalosis your carbon dioxide levels are low. This causes
your body to release more bicarbonate to return your blood pH level back
to normal. This is called compensated alkalosis. Your blood pH levels will
test normal, however your kidneys are releasing more bicarbonate,
compensating for the lower levels of carbon dioxide.

When your blood has too much bicarbonate, it is called metabolic alkalosis.
This can happen from prolonged vomiting. Prolonged vomiting can also
make you lose too much chloride. This is called hypochloremic alkalosis.
Some diuretic medicines can cause you to lose too much potassium. This
is called hypokalemic alkalosis.

Diagnosing alkalosis
Along with a physical exam, diagnostic testing for alkalosis may include a
metabolic panel, blood gas analysis, urinalysis, and urine pH.

Treatment for alkalosis

Some medications (such as chloride and potassium) can help correct

chemical losses. Further treatment will depend on the cause. Your
physician will need to monitor your vital signs and create a proper plan to
correct your pH imbalance.

Complications of alkalosis

In severe cases, alkalosis can lead to heart arrhythmias or coma.

GLUCONEOGENESIS?
Gluconeogenesis is the synthesis of new glucose molecules from
pyruvate, lactate, glycerol, or the amino acids alanine or glutamine. This
process takes place primarily in the liver during periods of low glucose, that
is, under conditions of fasting, starvation, and low carbohydrate diets.

Certain key organs, including the brain, can use only glucose as an energy
source; therefore, it is essential that the body maintain a minimum blood
glucose concentration. When the blood glucose concentration falls below
that certain point, new glucose is synthesized by the liver to raise the blood
concentration to normal.

Gluconeogenesis is not simply the reverse of glycolysis. There are some

important differences. Pyruvate is a common starting material for
gluconeogenesis.
First, the pyruvate is converted into oxaloacetate. Oxaloacetate then serves
as a substrate for the enzyme phosphoenolpyruvate carboxykinase
(PEPCK), which transforms oxaloacetate into phosphoenolpyruvate (PEP).
From this step, gluconeogenesis is nearly the reverse of glycolysis. PEP is
converted back into 2-phosphoglycerate, which is converted into 3-
phosphoglycerate.

Then, 3-phosphoglycerate is converted into 1,3 bisphosphoglycerate and

then into glyceraldehyde-3-phosphate. Two molecules of glyceraldehyde-3-
phosphate then combine to form fructose-1-6-bisphosphate, which is
converted into fructose 6-phosphate and then into glucose-6-phosphate.
Finally, a series of reactions generates glucose itself. In gluconeogenesis
(as compared to glycolysis), the enzyme hexokinase is replaced by
glucose-6-phosphatase, and the enzyme phosphofructokinase-1 is
replaced by fructose-1,6-bisphosphatase. This helps the cell to regulate
glycolysis and gluconeogenesis independently of each other.

As part of lipolysis, fats can be broken down into glycerol, which can be
phosphorylated to form dihydroxyacetone phosphate or DHAP. DHAP can
either enter the glycolytic pathway or be used by the liver as a substrate for
gluconeogenesis.

HMP SHUNT PATHWAY?

(REFER 5 MARK QUESTION FOR ANSWERS)

UREA CYCLE?

(REFER 5 MARK QUESTION FOR ANSWERS)

ENZYME INHIBITION?
Enzyme inhibitors are molecules or compounds that bind to enzymes and
result in a decrease in their activity. An inhibitor can bind to an enzyme and
stop a substrate from entering the enzyme's active site and/or prevent the
enzyme from catalyzing a chemical reaction. There are two categories of
inhibitors.

1. irreversible inhibitors

2. reversible inhibitors
Inhibitors can also be present naturally and can be involved in metabolism
regulation. For example. negative feedback caused by inhibitors can help
maintain homeostasis in a cell. Other cellular enzyme inhibitors include
proteins that specifically bind to and inhibit an enzyme target. This is useful
in eliminating harmful enzymes such as proteases and nucleases.

Reversible inhibitors can bind to enzymes through weak non-covalent

interactions such as ionic bonds, hydrophobic interactions, and hydrogen
bonds. Because reversible inhibitors do not form any chemical bonds or
reactions with the enzyme, they are formed rapidly and can be easily
removed; thus the enzyme and inhibitor complex is rapidly dissociated in
contrast to irreversible inhibition.
Examples of reversible inhibition:

o competitive inhibition (Raises Km only)

o uncompetitive inhibition (Lowers Vmax and Km)

o noncompetitive inhibition (Lowers Vmax only)

Examples of irreversible inhibition:

o group specific: reacts only to certain chemical group.

o reactive substrate analogs (affinity label): inhibitor that are

structurally similar to the substrate and will bind to active site.
 o mechanism-based inhibitors (suicide inhibitors): enzymes
converts the inhibitor into a reactive form within active site.
Competitive inhibition can be overcome by increasing the concentration of
substrate while uncompetitive and noncompetitive inhibition cannot.

There are three kinds of reversible

inhibitors: competitive, noncompetitive/mixed, and uncompetitive inhibitors.

 Competitive inhibitors, as the name suggests, compete with

substrates to bind to the enzyme at the same time. The inhibitor has an
affinity for the active site of an enzyme where the substrate also binds
to. This type of inhibition can be overcome by increasing the
concentrations of substrate, out-competing the inhibitor. Competitive
inhibitors are often similar in structure to the real substrate.

 Uncompetitive inhibitors bind to the enzyme at the same time as the

enzyme's substrate. However, the binding of the inhibitor affects the
binding of the substrate, and vice-versa. This type of inhibition cannot be
overcome, but can be reduced by increasing the concentrations of
substrate. The inhibitor usually follows an allosteric effect where it binds
to a different site on the enzyme than the substrate. This binding to an
allosteric site changes the conformation of the enzyme so that the
affinity of the substrate for the active site is reduced.

 Non-competitive inhibitors bind to the other sites (Allosteric Sites), not

the active site, and stops the enzyme's activity by changing the shape of
the active site (caused by disruption to the normal arrangement of
hydrogen bonds and weak hydrophobic interactions holding the enzyme
molecule together in its 3D shape. This distortion ripples to the active
site making it unsuitable) . Therefore, concentration of the substrate is
meaningless unlike in competitive inhibition.


Few examples of Reversible inhibitors:
Acetylcholinesterase inhbitors: Often abbreviated AChEI or anti-
cholinesterase it is a chemical that inhibits the
enzyme Acetylcholinesterase from breaking down acetylcholine. This
ultimately leads to increase in both the level and longevity of action of the
neurotransmitter acetylcholine.
Reversible inhibitor of monoanime oxidase A(maoA): maoA inhibitors
compromise of a wide range of natural as well as psychiatric drugs that
inhibits the enzyme monoamine oxidase temporarily and reversibly. maoA
inhibitors are most commonly used to fight depression and dysthymia.

Irreversible inhibitors
Irreversible inhibitors covalently bind to an enzyme, cause chemical
changes to the active sites of enzymes, and cannot be reversed. A main
role of irreversible inhibitors include modifying key amino acid residues
needed for enzymatic activity. They often contain reactive functional groups
such as aldehydes, alkenes, or phenyl sulphonates. These electrophilic
groups are able to react with amino acid side chains to form covalent
adducts.

Ketosis and Ketone bodies?

Ketosis is a metabolic state in which some of the body's energy supply
comes from ketone bodies in the blood, in contrast to a state of glycolysisin
which blood glucose provides energy. Generally, ketosis occurs when the
liver is metabolizing fatty acids or ethanol at a high rate in the absence of
glucose and converting the product acetyl-CoA into ketone bodies.
The first ketone body to be produced is acetoacetate, followed by
enzymatic reduction by beta-hydroxybutyrate dehydrogenase to beta-
hydroxybutyrate with NADH and a proton as a cofactor, or loss of a carbon
dioxide molecule to form acetone. As the liver itself lacks the metabolic
machinery required to utilize them, the product ketone bodies are entirely
released into the blood for use by the rest of the body.
Ketosis is a nutritional process characterised by serum concentrations of
ketone bodies over 0.5 mM, with low and stable levels of insulin and blood
glucose. It is almost always generalized with hyperketonemia, that is, an
elevated level of ketone bodies in the blood throughout the body. Ketone
bodies are formed by ketogenesis when liver glycogenstores are depleted
(or from metabolising medium-chain triglycerides). Ketones can also be
consumed in exogenous ketone foods and supplements.
The main ketone bodies used for energy are acetoacetate and β-
hydroxybutyrate, and the levels of ketone bodies are regulated mainly
by insulin and glucagon. Most cells in the body can use both glucoseand
ketone bodies for fuel, and during ketosis, free fatty acids and glucose
synthesis (gluconeogenesis) fuel the remainder.
Longer-term ketosis may result from fasting or staying on a low-
carbohydrate diet (ketogenic diet), and deliberately induced ketosis serves
as a medical intervention for various conditions, such as intractable
epilepsy, and the various types of diabetes.
In glycolysis, higher levels of insulin promote storage of body fat and block
release of fat from adipose tissues, while in ketosis, fat reserves are readily
released and consumed. For this reason, ketosis is sometimes referred to
as the body's "fat burning mode.
The difference between ketosis and ketoacidosis is the level of ketones in
the blood. Ketosis is a physiological adaptation to a low carbohydrate
environment like fasting or a ketogenic diet. There are situations (such
as treatment-resistant epilepsy) where ketosis can be beneficial to health.
Ketoacidosis is an acute life-threatening state requiring prompt medical
intervention; its most common form is diabetic ketoacidosis where both
glucose and ketone levels are significantly elevated.

The two sources of ketone bodies in the body are fatty acids in adipose
tissue and ketogenic amino acids. The main formation of ketone bodies is
through ketogenesis.
Adipose tissue can be used to store fatty acids for regulating temperature
and energy.These fatty acids can be released by adipokine signaling of
high glucagon and epinephrine levels, which inversely corresponds to
low insulin levels. High glucagon and low insulin correspond to times of
fasting or to times when blood glucose levels are low.[23] Fatty acids must
be metabolized in mitochondria in order to produce energy, but free fatty
acids cannot penetrate biological membranes due to their
negative electrical charge. So coenzyme A is bound to the fatty acid to
produce acyl-CoA, which is able to enter the mitochondria.
Once inside the mitochondrion, the dominant way that the bound fatty acids
are used as fuel in cells is through β-oxidation, which cleaves two carbons
off of the acyl-CoA molecule in every cycle to form acetyl-CoA. Acetyl-CoA
enters the citric acid cycle, where it undergoes an aldol
condensation with oxaloacetate to form citric acid; citric acid then enters
the tricarboxylic acid cycle (TCA), which harvests a very high energy yield
per carbon in the original fatty acid.
Acetyl-CoA can be metabolized through the TCA in any cell, but it can also
undergo a different process in liver cells: ketogenesis, which
produces ketone bodies.Ketone bodies are also produced in mitochondria,
and usually occur in response to low blood glucose levels.
When glucose levels are low, oxaloacetate is diverted away from the TCA
cycle and is instead used to produce glucose de novo (gluconeogenesis).
But when oxaloacetate is unavailable to condense with acetyl-CoA, acetyl-
CoA cannot enter the cycle, and so the body has evolved an alternative
way to harvest energy from it.
In ketogenesis, two acetyl-CoA molecules instead condense to
form acetoacetyl-CoA via thiolase. Acetoacetyl-CoA momentarily combines
with another acetyl-CoA via HMG-CoA synthase to form hydroxy-β-
methylglutaryl-CoA. Hydroxy-β-methylglutaryl-CoA form the ketone
body acetoacetate via HMG-CoA lyase. Acetoacetate can then reversibly
convert to another ketone body—D-β-hydroxybutyrate—via D-β-
hydroxybutyrate dehydrogenase.
Alternatively, acetoacetate can spontaneously degrade to a third ketone
body (acetone) and carbon dioxide, although the process generates much
greater concentrations of acetoacetate and D-β-hydroxybutyrate. When
blood glucose levels are low, ketone bodies can be exported from the liver
to supply crucial energy to the brain.
Along with the fatty acids, deaminated ketogenic amino acids can also be
converted into intermediates in the citric acid cycle and produce ketone
bodies.

Tyrosine Metabolism?
Tyrosine (symbol Tyr or Y) or 4-hydroxyphenylalanine is one of the 20
standard amino acids that are used by cells to synthesize proteins. It is
a non-essential amino acid with a polar side group. The word "tyrosine" is
from the Greek tyros, meaning cheese, as it was first discovered in 1846 by
German chemist Justus von Liebig in the protein casein from cheese.It is
called tyrosyl when referred to as a functional group or side chain. While
tyrosine is generally classified as a hydrophobic amino acid, it is more
hydrophilic than phenylalanine.

Metabolism

Phosphorylation and sulfation

Some of the tyrosine residues can be tagged (at the hydroxyl group) with a
phosphate group (phosphorylated) by protein kinases. In its phosphorylated
form, tyrosine is called phosphotyrosine. Tyrosine phosphorylation is
considered to be one of the key steps in signal transduction and regulation
of enzymatic activity.
Phosphotyrosine can be detected through specific antibodies. Tyrosine
residues may also be modified by the addition of a sulfate group, a process
known as tyrosine sulfation
Precursor to neurotransmitters and hormones
In dopaminergic cells in the brain, tyrosine is converted to L-DOPA by
the enzyme tyrosine hydroxylase (TH). TH is the rate-limiting
enzyme involved in the synthesis of the neurotransmitter dopamine.
Dopamine can then be converted into other catecholamines, such
as norepinephrine (noradrenaline) and epinephrine (adrenaline).
The thyroid hormones triiodothyronine (T3) and thyroxine (T4) in
the colloid of the thyroid also are derived from tyrosine.

Precursor to alkaloids
The latex of Papaver somniferum, the opium poppy, has been shown to
convert tyrosine into the alkaloid morphine and the bio-synthetic pathway
has been established from tyrosine to morphine by using Carbon-14 radio-
labelled tyrosine to trace the in-vivo synthetic route.
Precursor to natural phenols
Tyrosine ammonia lyase (TAL) is an enzyme in the natural phenols
biosynthesis pathway. It transforms L-tyrosine into p-coumaric acid.
Precursor to pigments
Tyrosine is also the precursor to the pigment melanin.
Role of coenzyme Q10 synthesis
Tyrosine (or its precursor phenylalanine) is needed to synthesize the
benzoquinone structure which forms part of coenzyme Q10.
Digestion and Absorption of Proteins?

From mouth to stomach

The first step in food digestion (or any other protein food) involves chewing.
The teeth begin the mechanical breakdown of the large pieces into smaller
pieces that can be swallowed. The salivary glands provide some saliva to
aid swallowing and the passage of the partially mashed
through the esophagus. The mashed pieces enter the stomach through the
esophageal sphincter. The stomach releases gastric juices containing
hydrochloric acid and the enzyme, pepsin, which initiate the breakdown of
the protein. The acidity of the stomach facilitates the unfolding of the
proteins that still retain part of their three-dimensional structure after
cooking and helps break down the protein aggregates formed during
cooking. Pepsin, which is secreted by the cells that line the stomach,
dismantles the protein chains into smaller and smaller fragments. Proteins
are large globular molecules and their chemical breakdown requires time
and mixing. The powerful mechanical stomach contractions churn the
partially digested protein into a more uniform mixture called chyme. Protein
digestion in the stomach takes a longer time than carbohydrate digestion,
but a shorter time than fat digestion. Eating a high-protein meal increases
the amount of time required to sufficiently break down the meal in the
stomach. Food remains in the stomach longer, making you feel full longer.

Protein digestion requires the chemical actions of gastric juice and the
mechanical actions of the stomach.
From Stomach to Small Intestine

The stomach empties the chyme containing the broken down pieces into
the small intestine, where the majority of protein digestion occurs. The
pancreas secretes digestive juice that contains more enzymes that further
break down the protein fragments. The two major pancreatic enzymes that
digest proteins are chymotrypsin and trypsin. The cells that line the small
intestine release additional enzymes that finally break apart the smaller
protein fragments into the individual amino acids. The muscle contractions
of the small intestine mix and propel the digested proteins to the absorption
sites. The goal of the digestive process is to break the protein into
dipeptides and amino acids for absorption.
In the lower parts of the small intestine, the amino acids are transported
from the intestinal lumen through the intestinal cells to the blood. This
movement of individual amino acids requires special transport proteins and
the cellular energy molecule, adenosine triphosphate (ATP). Once the
amino acids are in the blood, they are transported to the liver. As with other
macronutrients, the liver is the checkpoint for amino acid distribution and
any further breakdown of amino acids, which is very minimal. Recall that
amino acids contain nitrogen, so further catabolism of amino acids releases
nitrogen-containing ammonia.
Because ammonia is toxic, the liver transforms it into urea, which is then
transported to the kidney and excreted in the urine. Urea is a molecule that
contains two nitrogens and is highly soluble in water. This makes it a good
choice for transporting excess nitrogen out of the body. Because amino
acids are building blocks that the body reserves in order to synthesize other
proteins, more than 90 percent of the protein ingested does not get broken
down further than the amino acid monomers.
Very little protein makes it to the large intestine if you are not eating
excessive amounts. If you have smelly flatulence, this may be a sign you
are eating too much protein because the excess is making it to the colon
where you gut microbes are digesting it and producing smelly gas.

Protein Absorption

In adults, essentially all protein is absorbed as tripeptides, dipeptides or

amino acids and this process occurs in the duodenum or proximal jejunum
of the small intestine. The peptides and/or amino acids pass through the
interstitial brush border by facilitative diffusion or active transport. Active
transport sodium and ATP to actively transport the molecule through the
cell membrane.
The R group determines the type of transporter used. Once passed through
the membrane, the amino acids or peptides are released into the intestinal
blood stream and are transported to the liver by the hepatic (liver) portal
vein. This is known as the enterohepatic circulation.

In the liver, 50-65% remain and are used to synthesize protein, nitrogen
containing compounds and form purine/pyrimidine bases. In some cases,
they may be converted to energy. The liver regulates the amino acid levels
in the blood. The amino acids that do not stay in the liver, pass through and
are transported to the rest of the body to be taken up and utilized by other
cells. Most branch chain amino acids pass through the liver.

Nitrogen Metabolism Overview

Amino acids are unique because they contain nitrogen. Several things can
happen to the nitrogen. First, it can remain on the molecule and be
incorporated into the product that cell is making, for example, a
polypeptide.

The nitrogen may be transaminated, in other words, the amine group (NH 2)
is transferred to another carbon skeleton to form a new amino acid. An
example would be the transfer of the amine from the non-essential amino
acid, alanine, to alpha-ketoglutaric acid to make glutamic acid, another
non-essential amino acid. The water-soluble vitamin B 6 is needed for this
process.
The amine group may be removed from the amino acid in a process known
as deamination. This process is used for the excretion of the nitrogen, and
the carbon skeleton is used to produce energy. Again, vitamin B6 is needed
for this process.
The nitrogen removed from amino acids is excreted via several different
routes. The most familiar path is urine where most of the nitrogen is in the
form of urea. Nitrogen is also excreted in the feces, skin, hair, and nails. In
skin, hair, and nails the nitrogen is bound to protein as this is the building
block of each.
Amino Acids Are Recycled

Just as some plastics can be recycled to make new products, amino acids
are recycled to make new proteins. All cells in the body continually break
down proteins and build new ones, a process referred to as protein
turnover. Every day over 250 grams of protein in your body are dismantled
and 250 grams of new protein are built.
To form these new proteins, amino acids from food and those from protein
destruction are placed into a “pool.” Though it is not a literal pool, when an
amino acid is required to build another protein it can be acquired from the
additional amino acids that exist within the body. Amino acids are used not
only to build proteins, but also to build other biological molecules containing
nitrogen, such as DNA and RNA, and to some extent to produce energy. It
is critical to maintaining amino acid levels within this cellular pool by
consuming high-quality proteins in the diet, or the amino acids needed for
building new proteins will be obtained by increasing protein destruction
from other tissues within the body, especially muscle. This amino acid pool
is less than one percent of total body protein content. Thus, the body does
not store protein as it does with carbohydrates (as glycogen in the muscles
and liver) and lipids (as triglycerides in adipose tissue).

Vitamin – A?

Vitamin A is a group of unsaturated nutritional organic compounds that

includes retinol, retinal, retinoic acid, and
several provitamin A carotenoids (most notably beta-carotene). Vitamin A
has multiple functions: it is important for growth and development, for the
maintenance of the immune system and good vision.
Vitamin A is needed by the retina of the eye in the form of retinal, which
combines with protein opsin to form rhodopsin, the light-absorbing
molecule necessary for both low-light (scotopic vision) and color
vision.Vitamin A also functions in a very different role as retinoic acid (an
irreversibly oxidized form of retinol), which is an important hormone-
like growth factor for epithelial and other cells.
Vitamin A can be found in two principal forms in foods:

 Retinol, the form of vitamin A absorbed when eating animal food

sources, is a yellow, fat-soluble substance. Since the pure alcohol form
is unstable, the vitamin is found in tissues in a form of retinyl ester. It is
also commercially produced and administered as esters such as retinyl
acetate or palmitate.

 The carotenes alpha-carotene, beta-carotene, gamma-carotene; and

the xanthophyll beta-cryptoxanthin (all of which contain beta-
ionone rings), but no other carotenoids, function as provitamin A
in herbivores and omnivore animals, which possess the enzyme beta-
carotene 15,15'-dioxygenase which cleaves beta-carotene in the
intestinal mucosa and converts it to retinol.

Vitamin A Deficiency

Vitamin A deficiency can occur as either a primary or a secondary

deficiency. A primary vitamin A deficiency occurs among children and adults
who do not consume an adequate intake of provitamin A carotenoids from
fruits and vegetables or preformed vitamin A from animal and dairy
products. Early weaning from breastmilk can also increase the risk of
vitamin A deficiency.

Vitamin A deficiency is "the leading cause of preventable childhood

blindness,"

Adequate supply, but not excess vitamin A, is especially important for

pregnant and breastfeeding women for normal fetal development and in
breastmilk. Deficiencies cannot be compensated
by postnatal supplementation. Excess vitamin A, which is most common
with high dose vitamin supplements, can cause birth defects and therefore
should not exceed recommended daily values.

Over Consumption

Since vitamin A is fat-soluble, disposing of any excesses taken in through

diet takes much longer than with water-soluble B vitamins and vitamin C.
This allows for toxic levels of vitamin A to accumulate. These toxicities only
occur with preformed (retinoid) vitamin A (such as from liver). The
carotenoid forms (such as beta-carotene as found in carrots), give no such
symptoms, but excessive dietary intake of beta-carotene can lead
to carotenodermia, a harmless but cosmetically displeasing orange-yellow
discoloration of the skin.

Sources

Cod liver oil, liver turkey, liver beef, pork, fish, liver chicken, ghee, sweet
potato, carrot, broccoli leaf, butter, collard greens, butternut squash,
spinach, pumpkin, cheddar cheese, egg, papaya, mango, tomatoes, pea,
milk etc.

Functions
Vitamin A plays a role in a variety of functions throughout the body,[3] such
as:

 Vision

 Gene transcription

 Immune function

 Embryonic development and reproduction

 Bone metabolism

 Haematopoiesis

 Skin and cellular health

 Teeth

 Mucous membrane

Vitamin D?
Vitamin D is a group of fat-soluble secosteroids responsible for increasing
intestinal absorption of calcium, magnesium, and phosphate, and multiple
other biological effects. In humans, the most important compounds in this
group are vitamin D3 (also known as cholecalciferol) and vitamin
D2(ergocalciferol). Cholecalciferol and ergocalciferol can be ingested from
the diet and from supplements. Only a few foods contain vitamin D. The
major natural source of the vitamin is synthesis of cholecalciferol in the skin
from cholesterol through a chemical reaction that is dependent on sun
exposure (specifically UVB radiation).

Vitamin D from the diet, or from skin synthesis, is biologically inactive. A

protein enzyme must hydroxylate it to convert it to the active form. This is
done in the liver and in the kidneys. vitamin D can be synthesized in
adequate amounts by most mammals exposed to sufficient sunlight.

Vitamin D has a significant role in calcium homeostasis and metabolism. Its

discovery was due to effort to find the dietary substance lacking in children
with rickets (the childhood form of osteomalacia). Vitamin D supplements
are given to treat or to prevent osteomalacia and rickets.

Deficiency

A diet deficient in vitamin D in conjunction with inadequate sun exposure

causes osteomalacia (or rickets when it occurs in children), which is a
softening of the bones. In the developed world, this is a rare disease.

However, vitamin D deficiency has become a worldwide problem in the

elderly and remains common in children and adults.Low blood calcifediol
(25-hydroxy-vitamin D) can result from avoiding the sun. Deficiency results
in impaired bone mineralization and bone damage which leads to bone-
softening diseases, including rickets and osteomalacia. Being deficient in
vitamin D can cause intestinal absorption of dietary calcium to fall to
15%. When not deficient, an individual usually absorbs between 60-80%.

Rickets, a childhood disease, is characterized by impeded growth and soft,

weak, deformed long bones that bend and bow under their weight as
children start to walk. This condition is characterized by bow legs, which
can be caused by calcium or phosphorus deficiency, as well as a lack of
vitamin D; today, it is largely found in low-income countries in Africa, Asia,
or the Middle East and in those with genetic disorders such as
pseudovitamin D deficiency rickets.
Maternal vitamin D deficiency may cause overt bone disease from before
birth and impairment of bone quality after birth. Nutritional rickets exists in
countries with intense year-round sunlight such as Nigeria and can occur
without vitamin D deficiency.

Osteomalacia is a disease in adults that results from vitamin D deficiency.

Characteristics of this disease are softening of the bones, leading to
bending of the spine, bowing of the legs, proximal muscle weakness, bone
fragility, and increased risk for fractures. Osteomalacia reduces calcium
absorption and increases calcium loss from bone, which increases the risk
for bone fractures.

Sources
Although vitamin D is not present naturally in most foods, it is
commonly added as a fortification in manufactured foods. In some
countries, staple foods are artificially fortified with vitamin D
Over Consumption
Vitamin D toxicity is rare. It is caused by supplementing with high doses of
vitamin D rather than sunlight. The threshold for vitamin D toxicity has not
been established; however, according to some research, the tolerable
upper intake level (UL) is 4,000 IU/day for ages 9–71 (100 µg/day).