BIOCHEMISTRY

SHORT NOTES AND SHORT ANSWERS

1. Polysaccharide:
• Polysaccharides are polymers consisting of hundreds or thousands of monosaccharide
units.
• They are also called glycans or complex carbohydrates.
• They may be either linear, (e.g. cellulose) or branched, (e.g. glycogen) in structure.
• Polysaccharides have high molecular weight and are only sparingly soluble in water.
• They are not sweetish and do not exhibit any of the properties of aldehyde or ketone
group.
• Polysaccharides are of two types.
i. Homopolysaccharides (homoglycans) - composed of single kind of monasaccharide
ii. Heteropolysaccharides (heteroglycans) - composed of two or more different types of
monosaccharide.

Homoglycans:
1. Starch:
- Reserve carbohydrate of plant kingdom and is present abundantly in potatoes, tapioca,
rice, wheat.
- When starch is treated with boiling water, 10-20% is solubilised this part is called
amylose and the insoluble part is called amylopectin.
- Amylase: is a linear polymer of D-Glucose units joined by alpha- 1,4 glycosidic linkage
- Amylopectin: structurally similar to amylase but with side chains joining them by alpha-
1,6 linkage
2. Glycogen:
- Reserve carbohydrate in animals.
- Stored in liver and muscle
- Glycogen is composed of glucose units joined by alpha-1,4 and alpha-1,6 glycosidic
linkages.
- Molecular weight of glycogen is about 5 million Daltons.
- Glycogen is more branched and more compact than amylopectin.
- The branching points are made by alpha-1, 6 linkages.
3. Cellulose:
- Chief carbohydrate in plants
- Cellulose constitutes 99% of cotton, 50% of wood and 40% of straw, and is the most
abundant organic material in nature.
- It is made up of glucose units combined with cellobiose bridges or beta -1,4 linkages. It
has a straight line structure, with no branching points.
4. Inulin:
- It is clinically used to find renal clearance value and GFR.

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Heteroglycans:
1. Agar:
- It is prepared from sea weeds and contains galactose, glucose and other sugars.
- Agar is used as a supporting medium for electrophoresis and immunoelectrophoresis.
2. Mucopolysaccharide/ Glycosaminoglycans

2. Mucopolysaccharide/ Glycosaminoglycans:
- Mucopolysaccharides or glycosaminoglycans (GAG) are carbohydrates containing uronic
acid and amino sugars.
- Because of the presence of charged groups, they attract water molecules and so they
produce viscous solutions.
- Some examples of mucopolysaccharides are given below.
Hyaluronic Acid
It is present in connective tissues, tendons, synovial fluid and vitreous humour. It contains
glucosamine and glucuronic acid.
Heparin
It is an anticoagulant widely used when taking blood in vitro for clinical studies. It is also
used in vivo in suspected thromboembolic conditions to prevent intravascular coagulation. It
contains sulphated glucosamine.
Keratan Sulphate
It is the only GAG which does not contain any uronic acid. The repeating units are
galactose and N-acetyl glucosamine in beta linkage. It is found in cornea and tendons.

3. Complications of Diabetes Mellitus:

Acute Metabolic Complications:
1. Diabetic keto acidosis: Ketone body formation. As oxaloacetate is diverted for
gluconeogenesis, the TCA cycle cannot consume all the acetyl-CoA. Hence, more acetyl-
CoA is converted to ketone bodies. This leads to accumulation of ketone bodies in blood
(ketonemia). The presence of ketone bodies in urine (ketonuria) is assessed by Rothera
test. The accumulation of acidic ketone bodies lowers the plasma pH. So, metabolic acidosis
occurs. The condition is called diabetic ketoacidosis. Smell of acetone in the breath is
noticed. If not treated promptly and properly, the condition may be fatal. Patient may
become unconscious, comatose and die.

Chronic Complications of Diabetes Mellitus

1. Vascular diseases: Atherosclerosis in medium sized vessels, plaque formation and
consequent intravascular thrombosis may take place. If it occurs in cerebral vessels,
the result is paralysis. If it is in coronary artery, myocardial infarction results.
2. Complications in eyes: Early development of cataract of lens is due to the increased
rate of sorbitol formation, caused by the hyperglycemia. Retinal microvascular
abnormalities lead to retinopathy and blindness.
3. Neuropathy: leads to chronic kidney disease and renal failure.

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4.Glucose Tolerance Test:
• The patient who is scheduled for oral GTT is instructed to eat a high carbohydrate
diet for at least 3 days prior to the test, and come after an overnight fast on the day of
the test.
• A fasting blood glucose sample is first drawn.
• Then 75 gm of glucose (or 1.75 gm per kg body weight) dissolved in 300 ml of water
is given orally.
• After giving glucose, blood and urine specimens are collected at half hourly intervals
for at least 2 hours.
• Blood glucose content is measured and urine is tested for glycosuria.
• A curve is plotted for time against blood glucose concentration and is called glucose
tolerance curve.
Intravenous Glucose Tolerance Test
• The intravenous glucose tolerance test is often used for persons with malabsorptive
disorders or previous gastric or intestinal surgery.
• Glucose is administered intravenously over 30 minutes using 20% glucose solution. A
glucose load of 0.5 gm/kg of body weight is used.

Normal glucose tolerance curve

Normal response to glucose load is as follows:
• Initial fasting glucose is within the normal fasting limits (70 to 100 mg%).
• Blood glucose level rises to a peak (120 to 140 mg%) at half to 1 hour after ingestion of
glucose.
• The blood glucose level then returns rapidly to the fasting normal limits in about 2 hours.
• Glucose should not be present in any of the urine specimens collected for 2 hours.
Decreased glucose tolerance
Decreased glucose tolerance means decreased ability of the body to utilize glucose.
In decreased glucose tolerance:
• Fasting glucose is higher than normal limits.
• The blood glucose level rises above 180 mg/100 ml (renal threshold) after ingestion of
glucose.
• The blood glucose remains high for a longer time and may not return to fasting level
even after 3 hours.
• The urine samples corresponding to blood glucose level over 180 mg/100 ml may show
urine Benedict’s test positive (glycosuria).
• Decreased glucose tolerance occurs in diabetes mellitus and certain endocrine disorders
like:
– Hyperthyroidism
– Hyperpituitarism
– Hyperadrenalism (Cushing’s syndrome).
Increased glucose tolerance
Increased glucose tolerance means increased ability of the body to utilize glucose.
In increased glucose tolerance:

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• Fasting blood glucose is lower than normal.
• Only small rise in blood glucose level may be observed (not more than 100 mg%) even
after glucose administration.
• A flatter type of curve is obtained.
• No appearance of glucose in urine.
• This type of curve is obtained in endocrine
hypoactivity like:
– Hypothyroidism (myxedema, cretinism)
– Hypoadrenalism (Addison’s disease)
– Hypopituitarism.

5. Competitive and Non competitive inhibition:

Competitive Inhibition
i. The inhibitor molecules are competing with the normal substrate molecules for
attaching with the active site of the enzyme.
E+S —> E-S —> E+P
E+I —> E-I
ii. SinceE-I(enzyme–inhibitor complex) can react only to reform the enzyme and
inhibitor, the number of enzyme molecules available for E-S formation is reduced.
iii. In competitive inhibition, the inhibitor will be a structural analog of the substrate.
There will be similarity in three-dimensional structure between substrate (S) and
inhibitor (I). For example, the succinate dehydrogenase reaction is inhibited by
malonate, which are structural analogs of succinate
iv. Competitive inhibition is usually reversible. Excess substrate abolishes the
inhibition. If substrate concentration is enormously high when compared to inhibitor,
then the inhibition is reversed.
v. From the graphs, it is seen that in the case of competitive inhibition, the Km is
increased in presence of competitive inhibitor but Vmax is not changed. Thus,
competitive inhibitor apparently increases the Km.
Clinical Significance
i. Pharmacological action of many drugs may be explained by the principle of
competitive inhibition. A few important examples are given below:
ii. Sulphonamides are commonly employed anti- bacterial agents. Bacteria synthesise
folic acid by combining PABA with pteroyl glutamic acid. Bacterial wall is
impermeable to folic acid. Sulpha drugs, being structural analogs of PABA, will
inhibit the folic acid synthesis in bacteria, and they die. The drug is nontoxic to
human cells, because human beings cannot synthesise folic acid. Preformed folic acid
is essential for man.
iii. Methotrexate (4-amino-N10 -methyl folic acid) is a structural analog of folic acid, and
so can competitively inhibit folate reductase enzyme. This is essential for DNA
synthesis and cell division. Therefore, methotrexate is used as an anticancer drug.

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Noncompetitive Inhibition
i. A variety of poisons, such as iodo acetate, heavy metal ions (silver, mercury) and
oxidising agents act as irreversible noncompetitive inhibitors.
ii. The inhibitor usually binds to a different domain on the enzyme, other than the
substrate binding site.
iii. Since these inhibitors have no structural resemblance to the substrate, an increase in
the substrate concentration generally does not relieve this inhibition.
iv. Cyanide inhibits cytochrome oxidase. Fluoride will remove magnesium ions and
will inhibit the enzyme, enolase, and consequently the glycolysis.
v. The inhibitor combines with the enzymes and the reaction becomes irreversible.
vi. The velocity(Vmax) is reduced. But Km value is not changed, because the
remaining enzyme molecules have the same affinity for the substrate.
vii. Increasing the substrate concentration will abolish the competitive inhibition, but will
not abolish non-competitive inhibition.

6. Essentail Amino Acid:

Essential amino acids cannot be synthesized by the body and must, therefore, be essentially
supplied through the diet.
Ten amino acids, essential for humans include:
• Phenylalanine
• Valine
• Threonine
• Tryptophan
• Isoleucine
• Methionine
• Histidine
• Arginine
• Lysine
• Leucine.
The mnemonics PVT. TIM. HALL or L.VITTHAL (MP).
Among the ten essential amino acids; arginine and histidine are known as semi-essential
amino acids since these amino acids are synthesised partially in human body. Arginine and
histidine become essential in diet during periods of rapid growth as in childhood and
pregnancy.
An essential amino acid must be provided in the diet. An absence of an essential amino acid
from the diet impairs protein synthesis and generally causes negative nitrogen balance, i.e.
the total nitrogen losses in the urine, feces and sweat exceed the dietary nitrogen intake.

7. Jaundice:
When bilirubin in blood exceeds 1mg/dL is called hyperbilirubinemia and when it reaches
a certain concentration approximately 2.2 to 5 mg/dL it diffuses into the tissues. The skin
and sclera appear yellowish due to deposition of bilirubin in the tissues. This clinical
condition is called jaundice (French : jaune = yellow) or icterus.

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Classification
Jaundice is classified into a. Pre hepatic jaundice. b. Hepatic jaundice. c. Post hepatic
jaundice based on causes.
a. Pre hepatic jaundice :Haemolytic jaundice is the other name given to this condition. It is
mainly due to increased hemolysis and hence name. Excess hemolysis leads to
formation of excess bilirubin. But liver is unable to conjugate excess bilirubin. So
accumulation of unconjugated free bilirubin occurs and plasma free bilirubin level is
elevated. Increased hemolysis is seen in hemoglobinopathies, incompatible blood
transfusion, hereditary spherocytosis and in malaria.
b. Hepatic jaundice: It is also known as hepato cellular jaundice. Because liver cell
damage is the main cause for this type of jaundice. Viral infections, toxins, chemicals
damage liver cells. Hepatitis virus, mushroom poisons, chloroform, carbon tetra
chloride and phosphorus damage hepatocytes. Antibiotics use and in cirrhosis also
hepatocytes are damaged. Functions of damaged hepatocytes are impaired. So
hepatocytes may not be able to conjugate or secrete bilirubin though the bilirubin
production is normal. If conjugation is impaired plasma level of unconjugated bilirubin
is elevated. If secretion of conjugated bilirubin is affected its level in plasma is elevated.
Hence in hepatic jaundice both conjugated and free bilirubin levels are increased.
c. Post hepatic jaundice: It is also known as obstructive jaundice. Bile duct obstruction
causes this disease. Stones in gallbladder and cancer of head of pancreas cause bile duct
obstruction. Due to block in bile flow conjugated bilirubin secreted into bile returns to
blood. Hence conjugated bilirubin level is elevated in obstructive jaundice.
• van den Bergh Reaction
It is based on Ehrlich's reaction. It is used for measurement of plasma bilirubin. In
this reaction a purple red color is produced. It is due to coupling of bilirubin with
diazo reagent or diazotized sulphanilic acid.

8. Gout:
Gout : It is common disease of purine nucleotide degradation. Plasma uric acid level is
elevated which is characteristic sign of gout.
Symptoms : Deposition of uric acid crystals occurs in soft tissues because uric acid is less
soluble in aqueous environment of body fluids.Tophi is the name given to urate crystals that
are found in joints, cartilage of fingers and toes. Arthritic type gouty attacks occurs in
affected individuals.
Causes :
1. Over production of uric acid causes gout. Increased purine nucleotide production
leads to excessive uric acid production. It occurs in HGPRT ase deficiency, Hyper
active PRPP synthetase, leukaemia, von Gierke’s disease and polycythemia.
2. Impaired removal of uric acid by kidneys causes gout. It is called as renal gout. It
occurs due to defective uric acid transport in renal tubular cells and
glomerulonephritis.

9.Clinical significance of enzymes:

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• Certain enzymes are used:
• For the diagnosis of the disease
• As therapeutic agents
• As analytical reagents

Diagnostic Use of Enzymes

Enzymes are known as marker of cellular damage and their measurement in plasma
is used in the investigation of diseases of liver, heart, skeletal muscle, the biliary
tract and the pancreas.

Alanine transaminase (ALT)

• Alanine transaminase was known formerly as glutamate pyruvate
transaminase (GPT).
• The plasma ALT normal value for adult is 10 to 40 U/L.
• ALT level is elevated in liver diseases (viral or toxic hepatitis),
jaundice and cirrhosis of liver
Aspartate transaminase (AST)
• It was known formerly as glutamate oxaloacetate transminase (GOT).
• The plasma AST normal value for adults is 10 to 30 U/L.
• Increased AST level occurs after myocardial infarction. The plasma AST
level starts increasing after 6 to 8 hours after the onset of chest pain with peak
values 18 to 24 hours and the values fall to normal level by the fourth or fifth
day.
• It is moderately elevated in liver disease.
Alkaline phosphatase (ALP)
• ALP hydrolyzes organic phosphate at alkaline pH.
• Normal serum level for adults is 3-13 KA units/dl.
• It is elevated in certain bone and liver disease
• Very high levels may be noticed in obstructive jaundice, bone diseases such
as Paget’s disease, rickets, osteomalacia, carcinoma of bone and
hyperparathyroidism.

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 Acid phosphatase (ACP)
• It hydrolyzes phosphoric acid ester at pH 5 to 6.
• Normal serum value for ACP is 0.5 to 4 KA units/dL.
• Prostatic acid phosphatase enzyme is useful for the diagnosis and prognosis of
prostate cancer. ACP is therefore an important tumor marker.
Amylase
• It catalyzes hydrolysis of starch and glycogen.
• Normal serum value is 50-120 U/L.
• The activity of serum amylase is increased in acute pancreatitis.
• Elevated activity of amylase is also found in urine of the patient of acute pancreatitis.
• Increase in serum levels are also seen in chronic pancreatitis, mumps and
obstruction of pancreatic duct.

10. Phospholipids:
• These are made up of fatty acid, glycerol or other alcohol, phosphoric acid and
nitrogenous base.
• Phospholipids are the major lipid constituents of cell
membranes.
A. Phosphatidates
i. These are derivatives of phosphatidic acid which is the simplest phospholipid.
ii. Phosphatidic acid is made up of one glycerol to which two fatty acid residues are
esterified to carbon atoms 1 and 2. The 3rd hydroxyl group is esterified to a phosphoric
acid
B. Amphipathic Nature
Phospholipids in general are amphipathic, parti- cularly lecithin. They have both
hydrophobic and hydrophilic portion in their molecule
C. Biomembranes
The molecules align themselves to form monolayers with the polar heads pointing in one
direction and the nonpolar tails in the opposite direction.They also act as detergents and
emulsifying agents. In vivo, they act as pulmonary surfactants.
D. Liposomes
Liposomes are microscopic spherical vesicles. When mixed in water under special
conditions, the phospholipids arrange themselves to form a bilayer membrane which
encloses some of the water in a phospholipid sphere. Drugs, proteins, enzymes, etc. may be
encapsulated by the liposomes which act as carriers for these substances to target organs.
Liposomes have important applications in cancer chemotherapy, antimicrobial therapy, gene
therapy, vaccines and diagnostic imaging.

11. Significance of HMP shunt pathway:

Significance of the HMP Shunt Pathway
1. Tissues
The oxidative phase of shunt pathway is seen in organs where fatty acid or steroid synthesis
is taking place, such as liver, mammary glands, testis, ovary, adipose tissue and adrenal
cortex. It has also an important role in RBCs and lens of eye.
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2. Generation of Reducing Equivalents
The major metabolic role of the pathway is to metabolic role of the pathway is to provide
cytoplasmic NADPH for reductive biosynthesis of fatty acids, cholesterol and steroids
3. ErythrocyteMembrane
NADPH is required by the RBC to keep the glutathione in the reduced state. NADPH,
glutathione and glutathione reductase are required to preserve the integrity of RBC
membrane.
4. Lens of Eye
Maximum concentration of NADPH is seen in lens of eye. For preserving the transparency
of lens, NADPH is required.
5. Availability of Ribose
Ribose and deoxyribose are required for DNA / RNA synthesis. Ribose is also necessary for
nucleotide coenzymes. Reversal of nonoxidative phase is present in all tissues, by which
ribose could be made available.
6. What about ATP?
ATP is neither utilized nor produced by the HMP shunt pathway. Cells do not use the shunt
pathway for energy production.
7. GPD Deficiency
The enzyme glucose-6-phosphate dehydrogenase (GPD) may be deficient in some persons.
It is the most common enzyme deficiency seen in clinical practice. It is an X-linked
condition. It will lead to drug-induced hemolytic anemia. The deficiency is manifested
only when exposed to certain drugs or toxins, e.g. intake of antimalarial drugs like
primaquin and ingestion of fava beans (Favism). Sulpha drugs and furadantin may also
precipitate the hemolysis. This is characterized by jaundice and severe anemia. GPD
deficiency is reported from almost all regions of India.
8. Met-hemoglobinemia
GPD deficient persons will show increased met- hemoglobin in circulation, even though
cyanosis may not be manifested.

12. Genetic code:

The information needed to direct the synthesis of protein is contained in the mRNA in the
form of a genetic code.
Codons are a group of three adjacent bases that specify the amino acids of protein.
Characteristics of Genetic Code
• Number of codons: There are 64 possible codon sequences. Because four nucleotide
bases A, G, C and U are used to produce the three base codons, there are therefore 43 or 64
possible codon sequences.
• Stop or termination or nonsense codons: Three of the 64 possible nucleotide triplets,
UAA, UAG and UGA do not code for any amino acids, they are called nonsense codons
that normally signal termination of polypeptide chains. These nonsense codons are
arbitarily named amber, ochre and opal.
• The code is degenerate but unambiguous: As there are 61 codons for 20 amino acids,
one amino acid has more than one codon and the code is referred to as degenerate,
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indicating that there are redundancies. Although an amino acid may have more than one
codon, each codon specifies only one amino acid. Thus, the genetic code is unambiguous.
Degeneracy minimizes the deleterious effects of mutations.
• Codons that designate the same amino acid are called synonyms. Two amino acids
methionine (AUG) and tryptophan (UGC) each have only one codon. The remaining amino
acids have multiple codons, e.g. arginine is specified by six different codons (Figure 20.9).
• The code is almost universal: That is, the meaning of each codon is the same in almost
all known organisms. Exceptions to the universality of the genetic code are found in human
mitochondria, where the code:
– UGA codes for tryptophan instead of serving as a stop codon
– AUA codes for methionine instead of isoleucine
– CUA codes for threonine instead of leucine.
• The code is non-overlapping and without punctuation: During translation, the code is
read sequentially, without spacer bases, from a fixed starting point, as a continuous
sequence of bases, taken 3 at a time, e.g. A U G C U A G A C U U U is read as:
AUG / CUA / GAC / UUU without “punctuation” between the codons.

13. Glycogen storage diseases:

This is a group of genetic diseases, that result from a defect in an enzyme
required for either glycogen synthesis or degradation and characterized by
deposition of either normal or abnormal glycogen in the speci c tissues.

14. Isoenzyme:

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fi
Isoenzymes or isozymes are multiple forms (isomers) of the same enzyme that catalyze the
same biochemical reaction. Isoenzymes show different chemical and physical properties like
electrophoretic mobility and kinetic properties.
Not all enzymes have isoenzymes. In fact, it was found that only those enzymes, which are
active in polymeric form demonstrate isoenzyme. For example:
1. Lactate dehydrogenase (LDH)
2. Creatine kinase (CK) (formerly called creatine phosphokinase (CPK).
Lactate Dehydrogenase (LDH)
Lactate dehydrogenase is a tetrameric enzyme that catalyzes the oxidation of L-lactate to
pyruvate.
• LDH has five isoenzymes:
– LDH1 ,– LDH2 ,– LDH3 ,– LDH4, – LDH5.
• Since LDH is a tetramer, made up of two types of polypeptide M (muscle) type and H
(heart) type, five combinations are possible with varying ratios of two kinds of
polypeptides.
• Five isoenzymes of LDH can be detected by electrophoresis as they have different
electrophoretic mobilities.
• LDH1 is the fastest moving fraction towards the anode and LDH5 is the slowest moving
isoenzyme of LDH
• LDH1 predominates in cells of cardiac muscle, and erythrocytes and LDH5 is the most
abundant form in the liver and in skeletal muscle
Clinical Applications of LDH
The LDH isoenzyme analysis may be useful in the following clinical situations:
• Significant elevation of LDH1 and LDH2 (LDH1>LDH2) occurs within 24 to 48 hours after
myocardial infarction.
• Predominant elevation of LDH2
and LDH3 occur in leukemia.
LDH3 is the main isoenzyme
elevated due to malignancy of
many tissues.
• Elevation of LDH5 occurs after
damage to the liver or skeletal
muscle.

Creatine Kinase (CK)

Creatine kinase isoenzymes are dimer that are made up of two types of polypeptide chains,
which may be either M (muscle) type or B (brain) type, generating three isoenzymes.
Clinical Application
• CK1 may be elevated in neonates particularly in damaged brain or very low birth weight
newborn.

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• Increased level of CK2 in blood is characteristic of damage of heart tissue from
myocardial infarction because cardiac tissue is the only tissue which has the mixed MB
(CK2) isoenzyme.
• Elevated levels of CK3 in serum occur in all types of dystrophies and myopathies.

15. Plasma lipoprotein:

• Lipoproteins are large water soluble complexes formed by a combination of lipid and
protein that transport insoluble lipids through the blood between different organs and
tissues.
• Lipoproteins consist of a lipid core containing nonpolar triacylglycerol and cholesterol
ester surrounded by a single layer of amphipathic phospholipids and free cholesterol
molecules with some proteins, (apoprotein).
• The protein components are referred to as an apoprotein or apolipoprotein. There are four
major types of apolipoproteins designated by letters A, B, C and E with subgroups given
in Roman numerals I, II, III, etc.
• Classes of Lipoproteins
Lipoproteins have been categorized into four major classes according to their physical and
chemical properties. These are :
1. Chylomicrons
2. Very low density lipoproteins (VLDL)
3. Low density lipoprotein (LDL)
4. High density lipoprotein (HDL).
• These lipoprotein complexes contain different proportions of lipids and proteins. The
density of these lipoproteins is inversely proportional to triacylglycerol content. As the
density increases, the diameter of the particle decreases.
• Chylomicrons containing about 1 percent protein and 99 percent triacylglycerol have the
lowest density .
• While HDL containing 50 percent of protein and 50 percent of lipid have the highest
density.
• Triacylglycerol is the predominant lipid in chylomicrons and VLDL. Cholesterol is the
predominant lipid in LDL, whereas phospholipid is the predominant lipid in HDL.
16. Inborn errors of amino acid:
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16. Inborn errors of amino acid metabolism:
Maple syrup urine disease (MSUD) or branched chain keto aciduria
- It is an inborn error or branched chain amino acids namely leucine, isoleucine and valine
catabolism
- Biochemical cause
• Maple syrup urine disease is due to inherited defect in the branched chain α-keto acid
dehydrogenase. Due to this defect α-keto acids of leucine, isoleucine and valine
cannot be further metabolized. As a result, the branched chain amino acids, leucine,
isoleucine and valine, and their α-keto acids accumulate in blood, urine and CSF.
• α-keto acids impart a characteristic sweet odor to the urine of the affected individuals
which resembles with maple syrup or burnt sugar hence the name.
- Symptoms
• Maple syrup urine disease is characterized by vomiting, dehydration severe metabolic
acidosis and a characteristic maple syrup odor to the urine.
• If untreated, it leads to mental retardation, coma and even death within one year after
birth.
- Diagnosis
• Maple syrup disease is diagnosed by estimating increased levels of branched chain
amino acids and their keto acids in plasma and urine.
- Treatment
• Treatment involves replacing dietary protein by mixture of amino acids that contain
low or no leucine, isoleucine and valine.
• To monitor the effectiveness of the dietary treatment, plasma and urinary levels of
branched chain amino acids should be measured constantly.
PHENYLKETONURIA (PKU)
i. Deficiency of phenylalanine hydroxylase is the cause for this disease. It is a recessive
condition.

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 ii. The child is mentally retarded with an IQ in the range of 25–50. About 20% inmates
of lunatic asylum may have PKU.
iii. Agitation, hyperactivity, tremors and convulsions are often manifested.
Laboratory Diagnosis
i. Blood phenylalanine level is drastically increased.
ii. Ferric chloride test: Phenylketone (phenyl pyruvate), phenyllactate and phenylacetate
are excreted in urine.This could be detected by adding a drop of ferric chloride to the
urine. A transient blue-green color is a positive test.
Treatment of Phenylketonuria
i. Early detection is very important.About 5 units of IQ are lost for each 10 week delay
in starting the treatment.
ii. The treatment is to provide a diet containing low phenylalanine. Food based on
tapioca (cassava) will have low phenylalanine content.
iii. This special diet is to be continued till 5 years of age; by which time brain
development is completed. After that the child can have a normal diet.
Alkaptonuria:
- Alkaptonuria is an autosomal recessive condition. The metabolic defect is the deficiency
of homogentisate oxidase. This results in excretion of homogentisic acid in urine. There
is no mental retardation.
- Diagnosis: Urine becomes black on standing when it becomes alkaline. The
homogentisic acid is oxidized to black colored alkaptone bodies. Ferric chloride test
will be positive for urine. Benedict's test is strongly positive. Therefore, alkaptonuria
comes under the differential diagnosis of reducing substances in urine.

17. Liver functional test:

Classification of Liver Function Tests:
Liver function tests can be classified into five classes according to the function of the liver
as given below:
Tests based on excretory function
It includes measurement of:
• Serum bilirubin
• Urine bilirubin
• Urine bile salts
• Bromosulphophthalein (BSP) dye tests.
Tests based on detoxification function
It includes determination of:
• Blood ammonia and bilirubin
• Hippuric acid test.
Tests based on synthetic function
It includes determination of:
• Plasma proteins, albumins and globulins
• Prothrombin time.
Tests based on metabolic function
It includes:
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• Test related to carbohydrate metabolism
– Galactose tolerance test.
• Test related to lipid metabolism
– Determination of serum cholesterol and ratio of free to esterified cholesterol.
• Test related to protein metabolism
– Serum protein estimation
– Serum ammonia estimation.
Determination of serum enzymes
• Serum alanine transaminase (ALT)
• Serum aspartate transaminase (AST)
• Serum alkaline phosphatase (ALP).

Dye excretion test for excretory function

In addition to excreting bilirubin, the liver is capable of eliminating various dyes or drugs by
the same excretory pathway as bilirubin.
- Bromosulfophthalein excretion test
A 5% solution of BSP is injected intravenously (the dose is 5 mg/kg body wt) and a
sample of blood is tested 45 minutes later for percentage of injected dye remaining in the
blood.
- Clinical interpretation
• In normals, the retention of BSP at 45 minutes is less than 5%. Impairment of liver cell
function causes an increase in BSP retention.
Tests Based on the Detoxification Function
The liver is involved in the detoxification and removal of potentially hazardous substances
from the body. These may be endogenous, e.g. ammonia and bilirubin (discussed earlier) or
exogenous chemicals and drugs.
Hippuric acid test
Hippuric acid test is based on detoxicating function of the liver. The liver removes benzoic
acid by conjugating it with glycine to hippuric acid which is excreted in the urine.
In hippuric acid test, a dose of sodium benzoate is given either orally or intravenously and
the amount of hippuric acid excreted in a fixed time is determined.
For the oral test, the patient ingests 6 gm sodium benzoate dissolved in about 250 ml water.
Urine collections are made for the next 4 hours and the amount of hippuric acid excreted is
estimated.
- Clinical Interpretation
• In normal subjects, at least 3 to 3.5 gm hippuric acid should be excreted in the bile
during the 4 hours period.
• It is decreased in hepatitis, tumors, cirrhosis and obstructive jaundice.
• Excretion is normal in hemolytic jaundice
Determination of blood ammonia
• Liver detoxicates ammonia to form urea. In a liver disease, the ability to remove
ammonia may be impaired. The normal level of blood ammonia is 40-70 µg/100 ml of
blood.

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- Clinical Interpretation
• High blood levels of ammonia are found in acute hepatitis and cirrhosis.
Tests Based on Synthetic Function
Liver is the main source of synthesis of plasma proteins, e.g. albumin, globulin (except γ-
globulins which are synthesized in the reticuloendothelial system), blood clotting factors,
e.g. fibrinogen, prothrombin and factors V, VII, IX, X. Impaired function of liver results in
decreased protein synthesis.
- Determination of serum albumin and globulin
The normal concentrations of serum proteins are given below:
• Total serum protein : 6 to 8 gm/dl
• Serum albumin : 3.5 to 5.5 gm/dl
• Serum globulin : 2 to 3.5 gm/dl
• Albumin/globulin ratio :1.2:1 to 2.5:1.
- Clinical Interpretation
• Hypoalbuminemia may occur in hepatocellular disease, e.g. cirrhosis.
• Hyperglobulinemia may be present in chronic inflammatory disorders such as cirrhosis
and in infectious hepatitis.
• In advanced stages of liver disease, albumin is decreased and globulins are increased,
so that the A/G ratio may be reversed. The concentration of total globulins increases
due to a rise in the γ- globulins (synthesized by reticuloendothelial system and not by
the liver) to compensate for a possible fall in the α-globulins by liver.
Enzymes in Diagnosis of Liver Disease
Liver cells contain several enzymes. In liver damage, these enzymes are released into blood
and levels of these enzymes increase in blood.
Serum transaminases
Liver is the richest source of:
– Aspartate transaminase (AST) which is previously called serum glutamate oxaloacetate
transaminase (SGOT).
Alanine transaminase (ALT) which is previously called serum glutamate pyruvate
transaminase (SGPT).
• The normal range for these enzymes are as follows:
• – AST or SGOT = 4–17 IU/L
• – ALT or SGPT = 3–15 IU/L.
• Although, both AST and ALT are commonly thought of as liver enzymes because of
their high concentrations in liver, only ALT is markedly specific for liver since AST is
widely present in myocardium, skeletal muscle, brain and kidney and may rise in acute
necrosis of these organs besides liver cell injury.
- Clinical interpretation
• Alanine transaminase (ALT) estimations are useful in early diagnosis to evaluate
severity and prognosis of liver disease.
• In hepatitis, the levels of both these enzymes (ALT and AST) are increased, which go
in thousand units, usually 500 to 1500 IU/L.

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 • In obstructive jaundice also an increase occurs but usually does not exceed 200 to 300
IU/L.
• In hemolytic jaundice, the level of these enzymes are normal.
Alkaline phosphatase (ALP)
ALP is produced by many tissues, especially bone, liver, intestine and placenta and is
excreted in the bile. Elevation in activity of the enzyme can thus be found in diseases of
bone, liver and in pregnancy. In the absence of bone disease and pregnancy, there are
elevated ALP levels generally due to hepatobiliary disease. The normal level of ALP in
the plasma is 3-13 KA units/100 ml (King Armstrong units).
- Clinical interpretation
• The greatest elevation (3 – 10 times normal) occurs in obstructive jaundice. The
enzyme ALP is normally excreted through bile. Obstruction to the flow of bile, causes
regurgitation of enzyme into the blood resulting in increased serum concentration.
• Slight to moderate increase is seen in hepatitis and cirrhosis.
• Normal serum ALP values are found in hemolytic jaundice.

18. Lipid profile - Significance of cholesterol:

Lipid profile tests are used to estimate increased risk of cardiovascular disease which
includes measurement of:
1. Total serum cholesterol
2. Serum triglycerides
3. HDL cholesterol
4. LDL cholesterol.
Total Serum Cholesterol
Enzymatic method for estimation of cholesterol
• Commercially available cholesterol reagents commonly combine all enzymes and other
required components into a single reagent.
• The reagent usually is mixed with 3 µL to 10 µL aliquot of serum or plasma, incubated
under controlled conditions for color development and absorbance is measured at about 500
nm.
• The reagents typically use a bacterial cholesterol ester hydrolase to hydrolyze cholesterol
esters to cholesterol and fatty acids.
• The 3-OH group of cholesterol is then oxidized to a ketone derivative and H2O2 by
cholesterol oxidase.
H2O2, is then measured in a peroxidase catalyzed reaction that forms dye.
Normal values and interpretation
• The normal range for healthy young adults is 150-270 mg/dL.
• It may be lower in children.
• The concentration increases with age.
• The concentration in the women is generally somewhat lower than in men upto the
time of menopause but then increase and may exceed that in men of the same age.
Increased concentration

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 • The total concentration is increased in:
• – Hypothyroidism
– Uncontrolled diabetes mellitus
– Nephrotic syndrome
– Extrahepatic obstruction of the bile ducts
– Various hyperlipidemias.
• Long time elevated cholesterol concentration (more than 240 mg/dL) is a serious risk
factor for the development of coronary artery disease.
• Lowering of plasma cholesterol concentration reduces the incidence of coronary
heart diseases.
• National Cholesterol Education Program (NCEP) defined the levels of serum
cholesterol believed to be desirable, tolerable or a serious risk factor for
development of coronary artery disease. The report classifies total cholesterol
concentration which is applicable to all individuals over 20 years age and sex.
Decreased concentration
Hypocholesterolemia is usually present in:
• Hyperthyroidism
• Hepatocellular disease
• Certain genetic defects, e.g. abetalipoproteinemia.

19. Fluorosis:
Consumption of drinking water containing excess fluorine leads to fluorosis
i. Fluoride level more than 2 ppm will cause chronic intestinal upset, gastroenteritis, loss of
appetite and loss of weight.
ii. Levels more than 5 ppm cause mottling of enamel, stratification and discoloration of
teeth.
iii. A level more than 20 ppm is toxic, leading to alternate areas of osteoporosis and
osteosclerosis, with brittle bones. This is called fluorosis.
iv. Ingested fluoride accumulates in bones. It is a cumulative toxin.
v. In fluorosis, blood concentration of fluoride increases to 50 microgram/100 ml; whereas
normal value is 4 microgram/100 ml.
vi. Fluorosis is characterized by joint defects; especially genu valgum. Due to increased
breakdown of bone matrix, excretion of hydroxy- proline in urine is enhanced.
There are two types of fluorosis. They are
1. Dental fluorosis and 2. Skeletal fluorosis.
• Dental fluorsis : Molted teeth is major symptom. Due to loss of enamel teeth appears dull,
patches and cavities are produced on surface of teeth.
• Skeletal fluorosis : Knock knees or genu valgum is main skeletal deformity seen. Major
joints becomes stiff and painful. Neurological disturbances due to spine deformities also
occurs.
Incidence of Fluorosis
Nellore, Nalgonda and Prakasam districts of Andhra Pradesh and Patiala district of Punjab
are badly affected by fluorosis. 25 million people in India are suffering from fluorosis,
spread in 15 states of India. In the vicinity of irrigation dams, the water level in wells will
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come up, along with salts including fluoride. This has resulted in widespread fluorosis in
Punjab, Rajasthan, UP, Delhi, Andhra Pradesh, Karnataka and Tamil Nadu. Water from deep
subsoil wells will also contain higher fluoride level.
Certain salts used in paan supari also have large content of fluoride.
Fluoride rich sources are sea fish, cheese, tea and jowar.
Fluorosis is highly prevalent in areas where jowar is the staple diet.
Fluorinated toothpaste contains 3,000 ppm of fluoride. Even ordinary toothpaste contains
fluoride about 700 ppm.
Prevention of Fluorosis
Provide fluoride free water, restriction of intake of jowar, supplementation of vitamin C and
avoiding fluoride containing toothpaste

20. Mutation:
i. An alteration in the genetic material results in a mutation. This may be either gross, so
that, large areas of chromosome are changed, or may be subtle with a change in one
or a few nucleotides.
ii. Mutation may be defined as an abrupt spontaneous origin of new character
iii. Statistically, out of every 106 cell divisions, one mutation takes place.
Classification of Mutations
A point mutation is defined as change in a single nucleotide. This may be subclassified as
(a) substitution; (b) deletion and (c) insertion. All of them may lead to missense, nonsense
or frameshift effects.
• Substitution
Replacement of a purine by purine ( A to G or G to A) or pyrimidine by pyrimidine (T to C
or C to T) is called Transition mutation. If a purine is changed to a pyrimidine (e.g. A to C)
or a pyrimidine to a purine (e.g. T to G), it is called a transversion. The point mutation
present in DNA is transcribed and translated, so that the defective gene produces an
abnormal protein.
• Deletion
Deletions may be subclassified into:
i. Large gene deletions, e.g. alpha thalassemia (entire gene) or hemophilia (partial).
ii. Deletion of a codon, e.g. cystic fibrosis (one amino acid, 508th phenyl alanine is
missing in the CFTR protein.
iii. Deletion of a single base,whichwillgiverise to frameshift effect.
• Insertion
Insertions or additions or expansions are sub- classified into:
i. Single base additions, leading to frameshift effect.
ii. Trinucleotide expansions. In Huntington's chorea, CAG trinucleotides are repeated
30 to 300 times. This leads to a polyglutamine repeat in the protein. The severity of
the disease is increased as the number of repeats are more.
EXAMPLE: For example, HbS or sicklecell hemoglobin is produced by a mutation of the
beta chain in which the 6th position is changed to valine, instead of the normal glutamate.

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Here, the normal codon GAG is changed to GUG. HbS has abnormal electrophoretic
mobility and subnormal function, leading to sickle-cell anemia.
Mutagens and Mutagenesis
Any agent which will increase DNA damage or cell proliferation can cause increased rate of
mutations also. Such substances are called mutagens. X-ray, gamma-ray, UV-ray, acridine
orange, etc. are well known mutagens. The rate of mutation is proportional to the dose of
irradiation

21. Dietary fibers and their role in human nutrition:

• Dietary fibre is mainly of plant origin. Indigestible plant polysaccharides like cellulose,
hemicellulose, pectin, gums and lignin constitutes dietary fibre. Importance of dietary
fibre in human health is recently recognized. Dietary fibre has several protective,
preventive and curative effects. It is required for good health.
• The incidence of colonic diseases like ulcerative colitis, piles, constipation and irritable
bowel syndrome decreases with use of fibre in the diet. Metabolic diseases like diabetes
mellitus, obesity, coronary artery disease, hypertension etc incidence decreases with use
of fibre diet. Dietary fibre lowers blood glucose, cholesterol and triglyceride levels.
However absorption of glucose, cholesterol and some minerals is slow in presence of
dietary fibre

22. Structure and functions of tRNA:

tRNA Function
• tRNA plays an important role in protein synthesis.
• It acts as an adapter molecule for linking amino acids to its specific codon present in
mRNA.
• Aminoacylation of tRNA is the first step in protein synthesis.
• tRNA is specific to each amino acid and carries them during the translation process in
the ribosomal subunits.

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 • The tRNA transfers the amino acid to the growing polypeptide chain in the
ribosomes, which has three binding sites for tRNA, namely A, P and E, which
correspond to aminoacyl, peptidyl and exit, respectively.
• This decoding of codons of mRNA by specific tRNAs continues until the entire
sequence for a polypeptide chain is translated.

23. Wald’s visual cycle:

i. Wald was awarded Nobel prize in 1967, for identifying the role of vitamin A in
vision. Rhodopsin is a membrane protein found in the photoreceptor cells of the
retina. Rhodopsin is made-up of the protein opsin and 11-cis- retinal.
ii. When light falls on the retina, the11-cis-retinal isomerises to all-transretinal (Fig.
16.3).
iii. Generation of Nerve Impulse: In visual pigments, the 11-cis retinal locks opsin in its
inactive form. The isomerisation and photo-excitation leads to generation of the nerve
impulse. This is a G-protein coupled reaction.
iv. A single photon can excite the rod cell. The photon produces immediate
conformational change in rhodopsin and all-transretinal is produced. The all-
transretinal is then released from the opsin protein.
Regeneration of 11-cis-retinal
i. After dissociation, opsin remains in retina; but transretinal enters the blood circulation.
The all-transretinal is isomerised to 11-cis-retinal in the retina itself in the dark by the
enzyme retinal isomerase. The 11-cis
retinal can recombine with opsin to
regenerate
rhodopsin.
ii. Alternatively, all-transretinal is
transported to liver and then reduced to
all-transretinol by alcohol
dehydrogenase (ADH), an NADH
dependent enzyme. ADH contains zinc,
and therefore, zinc is important in
retinol metabolism. The all-trans-retinol
is isomerized to 11-cis-retinol and then
oxidized to 11-cis- retinal in liver. This
is then transported to retina. This
completes the Wald's visual cycle

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Dark Adaptation Mechanism
i. Bright light depletes stores of rhodopsin in rods. Therefore when a person shifts
suddenly from bright light to a dimly lit area, there is difficulty in seeing, for
example, entering a cinema theater. After a few minutes, rhodopsin is resynthesized
and vision is improved. This period is called dark adaptation time.
ii. It is increased in vitamin A deficiency. Red light bleaches rhodopsin to a lesser extent;
so doctors use red glasses, during fluoroscopic X-ray examination of the patients.
Rods are for Vision in Dim Light
In the retina, there are two types of photosensitive cells, the rods and the cones. Rods are
responsible for perception in dim light. It is made up of 11-cis- retinal + opsin. Deficiency
of cis-retinal will lead to increase in dark adaptation time and night blindness.
Cones are for Color Vision
i. Cones are responsible for vision in bright light as well as color vision. They contain
the photosensitive protein, conopsin (photopsin).
ii. In cone proteins also, 11-cis-retinal is the chromophore. Reduction in number of
cones or the cone proteins, will lead to color blindness.

24. Structural organization of Proteins:

Primary structure of proteins :
1. Aminoacid sequence of a protein is known as primary structure of protein.
2. Peptide bonds and disulfide bonds are involved in primary structure.
Primary structure of insulin:
1. It consist of two polypeptide chains.
2. They are A chain and B chain.
3. Inter chain di sulfide bonds links two chains.
4. Further an intra chain disulfide bond is present in A chain.
5. Glycine is the N-terminal aminoacid and aspargine is the C-terminal amino acid in A
chain.
6. In the B chain alanine is C-terminal amino acid and phenyl alanine is the N-terminal
aminoacid.
Secondary structure of protein:
1. Two dimentional folding of polypeptide chain is known as secondary structure of
protein.
2. The folding of protein chain can be ordered or disordered.
3. The ordered secondary structures are α-helix and β-pleated sheet.
4. The disordered secondary structures are random coil and reverse turn or β-turn.
Alpha (α)Helix
It is the secondary structure found in α-Keratin of hair, nails and epidermis of the skin.
• Structural features of α-helix:
• Coiling of polypeptide or protein chain along long axis produce α- helix.
• α-helix is stabilized by intra chain hydrogen bonding.
• Peptide bonds are involved in hydrogen bonding.
• C=O and –N-H groups of peptide bond participate in hydrogen bonding.
There are 3. 6 aminoacids in one turn of α-helix.
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 • Peptide bonds that are four aminoacid residues away participate in hydrogen bonding
i. e. -N-H of second aminoacid residue and –C=O of sixth aminoacids are involved in
hydrogen bonding.
• α-helix of fibrous proteins is right handed.
• α-helix is destabilized by hydrophobic aminoacids.
• In contrast aromatic aminoacids stabilizes α-helix.
• α-helical regions are found in several other proteins.
Beta (β)Pleated Sheet
When two or more polypeptide chains line side by side along long axis beta pleated sheet is
formed. Adjacent segments of a protein or polypeptide chain may also form secondary
structure.
Structural features of β-Pleated Sheet
• Polypeptide chains are fully extended along long axis in beta pleated sheet.
• Inter chain hydrogen bonds stabilizes beta pleated sheet.
• Based on direction β-pleated sheet is divided into
i. Antiparallel β-pleated sheet and ii. Parallel β-pleated sheet.
• In antiparallel β-pleated sheet polypeptide chains run in opposite direction.
• In parallel β-pleated sheet polypeptide chains run in same direction.
• Pleated sheet is found in many proteins. Albumin and hemoglobin of blood contains
β- pleated sheet.
• Antiparallel β pleated sheet is found in β-Keratin of silk fibroin, spider web and
amyloid protein found in the brain of Alzheimer's disease patients.
• β-pleated sheet content varies among proteins.
Tertiary structure of protein:
1. It is formed due to three dimentional folding of polypeptide chain of protein in space.
2. Tertiary structure of protein contains ordered and disordered secondry structures i. e.
α-Helix, β-pleated sheet, random coil conformation etc.
Forces involved in maintenance:
1. Several non covalent bonds stabilizes tertiary structure.
2. Usually it refers to native conformation of a protein.
3. Internal hydrogen bonds, electrostatic, hydrophobic and van der Waals interactions
are bonds that keep tertiary structure intact.
4. In the case of proteins that are made up of only one polypeptide chain tertiary
structure is the final level of protein structure.
Quaternary structure of protein:
1. Proteins which are made up of more than one polypeptide chain contains quaternary
structure.
2. Such proteins are known as
oligomeric proteins and
constituent polypeptide chains
are referred as sub units or
protomers.

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25. Electron transport chain:
i. In the electron transport chain, or respiratory chain, the electrons are transferred from
NADH to a chain of electron carriers. The electrons flow from the more
electronegative components to the more electropositive components.
ii. All the components of electron transport chain (ETC) are located in the inner
membrane of mitochondria. There are four distinct multi- protein complexes.
iii. Complex I is also called NADH-CoQ reductase or NADH dehydrogenase complex.
Complex II is also named as Succinate-Q- Reductase. Complex III is known as
Cytochrome Reductase. Complex IV is Cytochrome Oxidase
iv. These are connected by two mobile carriers, coenzyme Q (CoQ) and cytochrome c.
CoQ connects complex II and III. Cytochrome c is in between complex III and IV.

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26. Structure and function of DNA:
Deoxyribonucleic acid (DNA) is composed of four deoxy ribonucleotides, i.e.
deoxyadenylate (A), deoxyguany- late (G), deoxycytidylate (C), and deoxythymidylate (T).
These units are combined through 3' to 5' phosphodiester bonds to polymerize into a long
chain. The nucleotide is formed by a combination of base + sugar + phosphoric acid. The 3'-
hydroxyl of one sugar is combined to the 5'-hydroxyl of another sugar through a phosphate
group
Watson-Crick Model of DNA Structure
The salient features of Watson-Crick model of DNA are given below (Figs 24.2 and 24.3):
1. Right Handed Double Helix
DNA consists of two polydeoxy ribonucleotide chains twisted around one another in a right
handed double helix similar to a spiral staircase. The sugar and phosphate groups comprise
the handrail and the bases jutting inside represent the steps of the staircase. The bases are
located perpendicular to the helix axis, whereas sugars are nearly at right angles to the axis.
2. The Base Pairing Rule
Always the two strands are complementary to each other. So, the adenine of one strand will
pair with thymine of the opposite strand, while guanine will pair with cytosine. The base
pairing (A with T; G with C) is called Chargaff's rule, which states that the number of
purines is equal to the number of pyrimidines.
3. Hydrogen Bonding
The DNA strands are held together mainly by hydrogen bonds between the purine and
pyrimidine bases.
There are two hydrogen bonds between A and T while there are three hydrogen bonds
between C and G.
4. Antiparallel
The two strands in a DNA molecule run antiparallel, which means that one strand runs in
the 5' to 3' direction, while the other is in the 3' to 5' direction. Other features are:
i. The spiral has a pitch of 3.4 nano meter sperturn.

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 ii. ii. Within a single turn, 10 base pairs are seen.Thus, adjacent bases are separated by
0.34 nm.

27. Iron:
1. Distribution of Iron
Total body iron content is 3 to 5 gm, 75% of which is in blood. Iron is present in almost all
cells.
Heme containing proteins are hemoglobin, myoglobin, cytochromes, cytochrome oxidase,
catalase. Non-heme iron containing proteins are transferrin, ferritin, hemosiderin.
Blood contains 14.5 g of Hb per 100 ml. About 75% of total iron is in hemoglobin, and 5%
is in myoglobin and 15% in ferritin. Normal iron kinetics are shown in Figure 18.5.
2. Requirement of Iron (ICMR, 1990)
i. Daily allowance for iron for an adult Indian is 20 mg of iron, out of which about 1–2
mg is absorbed. In Western countries, requirement is less (15 mg/day) because the
diet does not contain inhibitory substances.
ii. Pregnant women need 40 mg/day. Transfer of iron and calcium from mother to fetus
occurs mainly in the last trimester of pregnancy. Therefore during this period,
mother's food should contain surplus quantities of iron and calcium.
iii. In the first 3 months of life, iron intake is negligible because milk is a poor source of
iron. During this time, child is dependent on the iron reserve received from mother
during pregnancy. In premature babies, the transplacental transfer of iron might not
have taken place. Hence, such babies are at a risk of iron deficiency. After 3 months
of life, diet supplementation with cereals is essential for supplying the iron
requirement.
3. Sources of Iron
i. Leafy vegetables are good sources.
ii. Cereals contain moderate quantity of iron. In a typical Indian diet, the major quantity
of iron is received from cereals because of the bulk quantity.
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 iii. Liver and meat contain moderate quantities.
iv. Jaggery is a good source for iron.
v. Cooking in iron vessels will help to get iron.
vi. Milk is a very poor source of iron.
4. Factors Influencing Absorption of Iron
i. Reduced form of iron: Only Fe++ (ferrous) form (reduced form) is absorbed. Fe+++
(ferric) form is not absorbed.
ii. Ascorbic acid: Ferric ions are reduced with the help of gastric HCl, and ascorbic acid.
Therefore these will favour iron absorption.
iii. Interfering substances: Iron absorption is decreased by phytic acid (in cereals) and
oxalic acid (in leafy vegetables) by forming insoluble iron salts. Calcium, copper, lead
and phosphates will inhibit iron absorption.
5. Storage of Iron
The storage form is ferritin. It is seen in intestinal mucosal cells, liver, spleen and bone
marrow. In iron deficiency anemia, ferritin content is reduced.

IRON DEFICIENCY ANEMIA

Causes for Iron Deficiency
i. Nutritional deficiency of iron.
ii. Hookworm infection: This may be the most important cause, especially in rural
areas, where sanitation is poor.
iii. Repeated pregnancies: About 1 g of iron is lost from the mother during one delivery.
iv. Chronic blood loss: Hemorrhoids (piles), peptic ulcer, uterine hemorrhage.
Manifestations of Iron Deficiency Anemia
i. Iron deficiency is characterized by microcytic hypochromic anemia. Anemia results
when hemoglobin level is less than 12 g/dl.
ii. When the level is lower than 10 g, body cells lack oxygen and patient becomes
uninterested in surroundings (apathy). All metabolic processes become sluggish.
Anemia will lead to impaired attention, irritability, lowered memory and poor
scholastic performance. Anemia and apathy go hand in hand.
Iron Toxicity
i. Hemosiderosis: Iron excess is called hemosiderosis. Hemosiderin pigments are
golden brown granules, seen in spleen and liver. Hemosiderosis occurs in persons
receiving repeated blood transfusions. Here the regulation at the level of intestine is
circumvented leading to iron overload.
ii. Primary Hemosiderosis: It is also called hereditary hemochromatosis. In these
cases, iron absorption is increased and transferrin level in serum is elevated. Excess
iron deposits are seen.
iii. Hemochromatosis: When total body iron is higher than 25–30 g, hemosiderosis is
manifested. In the liver, hemosiderin deposit leads to death of cells and cirrhosis.
Pancreatic cell death leads to diabetes. Deposits under the skin cause yellow-brown
discoloration, which is called hemochromatosis. The triad of cirrhosis,
hemochromatosis and diabetes are referred to as bronze diabetes.

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28. Define glycolysis. Enumerate how glucose is completely oxidised:
i. Glycolysis is derived from the
Greek word, glykys = sweet; and
lysis = splitting. In this pathway
glucose is converted to pyruvate
(aerobic condition) or lactate
(anaerobic condition), along with
production of a small quantity of
energy.
ii. All the reaction steps take place in
the cytoplasm. It is the only
pathway that is taking place in all
the cells of the body.
iii. Glycolysis is the only source of
energy in erythrocytes.
iv. In strenuous exercise, when
muscle tissue lacks enough
oxygen, anaerobic glycolysis
forms the major source of energy
for muscles.
v. The glycolytic pathway may be
considered as the preliminary step
before complete oxidation.
vi. The glycolytic pathway also
provides carbon skeletons for
synthesis of certain nonessential
amino acids as well as glycerol
part of fat.
vii. Most of the reactions of the
glycolytic pathway are reversible,
which are also used for
gluconeogenesis.

Complete oxidation of glucose: Pyruvate is later oxidatively decarboxylated to acetyl-

CoA, which enters into the citric acid cycle. Complete oxidation of glucose through
glycolysis plus citric acid cycle will yield a net 32 ATPs

29. Function, sources and diseases of thiamine:

Sources
Cereals (whole wheat flour and unpolished handpound rice) are rich sources of thiamine.
When the grains are polished, aleurone layer is usually removed. Yeast is also a very good
source. Structure of thiamine is shown in Figure 17.1.
Physiological Role of Thiamine

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 i. Pyruvate dehydrogenase: The coenzyme form is thiamine pyrophosphate (TPP). It
is used in oxidative decarboxylation of alpha keto acids, e.g. pyruvate decarboxylase,
a component of the pyruvate dehydrogenase complex. It catalyzes the breakdown of
pyruvate, to acetyl-CoA, and carbon dioxide.
ii. Alpha ketoglutarate dehydrogenase: An analogous biochemical reaction that
requires TPP is the oxidative decarboxylation of alpha ketoglutarate to succinyl
CoA and CO2
iii. Transketolase in the hexose mono-phosphate shunt pathway of glucose
iv. The main role of thiamine (TPP) is in carbohydrate metabolism. So, the requirement
of thiamine is increased along with higher intake of carbohydrates.

Deficiency Manifestations of Thiamine

• Beriberi: Deficiency of thiamine leads to beriberi. The early symptoms are anorexia and
weakness.
• Wet Beriberi: Here cardio vascular manifestations are prominent. Edema of legs, face,
and serous cavities are the main features. Death occurs due to heart failure.
• Dry Beriberi: In this condition, CNS manifestations are the major features. Peripheral
neuritis with sensory disturbance leads to complete paralysis.
• Wernicke-Korsakoff syndrome: It is also called as cerebral beriberi. Clinical features
are those of encephalopathy plus psychosis. It is seen only when the nutrition is severely
affected.
• Polyneuritis: It is common in chronic alcoholics. Alcohol utilization needs large doses of
thiamine. Polyneuritis may also be associated with pregnancy and old ag

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