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Biochemistry and Clinical Pathology

In Easy English
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Contents
1 Introduction To Biochemistry 3
2 Carbohydrate 5
3 proteins 12
4 Lipids 18
5 Nucleic Acid 24
6 Enzymes 28
7 Vitamins 33
8 It is provided separately
9 Minerals 42
10 Water and Electrolytes 47
11 Introduction to Biotechnology 50
12 Organ Function Test 51
13 Introduction to Pathology of Blood and Urine 56

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Biochemistry And Clinical Pathology Chapter 1

Introduction To Biochemistry
Introduction To Biochemistry
The Word Biochemistry is formed by Bio means living , and chemistry means chemical
reaction or chemical molecule .
so we can say:
Biochemistry is a branch of science in which we study about chemical reaction or
biomolecule inside the living organism .

Biomolecules
The cell of living organism is composed of a few elements like C , H, O ,and N etc. are called
biomolecules .
More than 10,000 or 100,000 biomolecules are present in human body but some them
recognised . example
1) amino acids 2) monosaccharide 3) nucleotide

Scope Of Biochemistry In Pharmacy

1) Drug Formulation : The biochemistry helps in drug formulation .
2) Drug Improvement : The biochemistry helps in the improvement of drug .
3) half Life : The biochemistry helps to know the half life of a drug , and biochemical test is
performed to check how long a drug is stable on a given temperature .
4) Drug Storage : The storage condition required for a drug is estimated by biochemical test
.
5) Expiry : The biochemistry helps to know the expiry of a drug .

Cell and Its biochemical organisation

Cell
Cell is the smallest structural and functional unit of living organism , About 100 trillion cells
are found in a normal human being .

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Biochemical organisation of Cell
As the tissues are made up of cells , in the same way cells are made up of molecules (
biomolecules ) . In other words we can say as cells are building blocks of tissues ,
biomolecules are building blocks of cells .

Biomolecules
Biomolecules are those elements( like C,H, O , N) which make different type of chemical
compounds and structures .
More than 10,000 or 100,000 biomolecules are present in human body but some them
recognised .
when different small organic compounds like amino acids , monosaccharide , nucleotides
get together and form chemical compounds or structures called complex Biomolecules ,
major complexx biomolecules are following
1) Carbohydrate 2) Protein 3) lipids 4) Nucleic Acid

Biomolecules Building Blocks Major Functions

Protein Amino acids Form basic frame work of cell
DNA Deoxyribonucleotides Carry the genetic material
RNA Ribonucleotides Carry information for protein
synthesis
Carbohydrates Monosaccharide It stored in the form of glycogen
and
fulfill short term energy
requirement
Lipids Fatty acids glycerol It is second major source of
energy and fulfill long term
requirement .

Micromolecules : small molecule of low molecular weight , we can say monomers are
micromolecules .
Macromolecules : Large molecules with high molecular weight are Macromolecules , or we
can say polymers are macromolecules .

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Biochemistry And Clinical Pathology Chapter 2

Carbohydrate
Definition
Carbohydrates are organic substances which have C ,H , O atoms .and H and O are in the
ratio 2:1 as found in H2 O , with general formula Cn ( H2 O)n .
Carbohydrates have following Functional groups .
1) Alcoholic hydroxyl group --OH
2) Aldehyde Group --CHO
3) Ketone Group C O
Carbohydrates are classified in three ways :
1) on the basis of complexity
2) on the basis of functional groups
3) On the basis of reactivity .

Classification Of Carbohydrates
Carbohydrates

On the basis Of complexity On the basis of functional group On the basis of reactivity
( aldose , ketose ) ( reducing and non reducing sugar )

Monosaccharide oligosaccharide Polysaccharide

( 1 sugar ) ( 2 - 10 sugar ) ( more than 10 sugars )

Monosaccharides ( simple sugar )

Monosaccharides are basic unit of carbohydrates and can not be broken down by
hydrolysis into other simple sugar . example Glucose fructose and galactose .

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Monosaccharide is derived from Greek word "mono " which means single and "saccharide"
which means sugar .
They have a sweet flavour . They are water soluble . they have a crystalline appearance .

Structure Of Glucos ,Galactose and fructose(Open / straight/ Chain or Fisher Projection)

Closed / Ring Chain or Haworth Projection

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α & ß Glucose

D & L Glucose

( D - glucose )

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Olygosaccharides
Oligosaccharides are a type of carbohydrates , these are made up of 2-10 units of
monosaccharides . it can be further classified In disaccharides , trisaccharides etc.

Disaccharides
Disaccharides are a type of carbohydrate and made of tow monosaccharides , and on
hydrolysis it gives on molecule of glucose and one molecule of fructose .
Example

1) Maltose 2) lactose 3) sucrose .

Structures of maltose ( Glucose + Glucose )

Structure of Lactose ( galactose + Glucose )

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Structure Of Sucrose ( Glucose + fructose )

Polysaccharide
Polysaccharide is a type of carbohydrate and it is made up of more than 10 monosaccharide
units . example Starch , glycogen .
they do not have a sweet flavour .
They are water insoluble
They are carbohydrate with high molecular weight

Chemical Nature Of Starch

1)Glucose is stored in plants in the form of starch .
2) It is composed of two components " Amylose "and " Amylopectin"
3) Amylose is made up of 250 - 300 glucose , that are joined together by α -1 , 4 glycosidic
bond .
4) Amylose chains are unbranched and coiled .
5) Amylose are consist about 15 - 20 %
6) Amylopectin is a glucose polymer with α 1, 4 glycosidic linkage
7) side chains with about 12 glucose are linked to the main chain with α -1,6 glycosidic
linkage .

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Chemical Nature Of Glycogen
1) Glucose is stored in the form of glycogen in animals .
2) It is found mostly in liver and muscles .
3) It is often called animal starch .
4) It converted in to glucose and provide energy in short term requirement .
5) It is a polymer which is highly branched , and 8 - 10 glucose units present per branch .
6) The chain of glycogen are joined together by α 1, 4 glycosidic link .
7) and branches are joined to the main chain by α 1, 6 glycosidic bond .
8) The structure of glycogen is similar to amylopectin but it is more highly branched .

Reducing And Non - Reducing sugar

Reducing Sugar : All those carbohydrates which have a free aldehyde or ketonic group and
reduce Fehling' solution and Tollen's reagent are called reducing sugar . e.g. Glucose .
Non Reducing Sugar : The sugar which does not have free aldehyde or ketonic group is
called non reducing sugar . e.g. sucrose
Chemical properties Of carbohydrates
1 ) when Carbohydrates are heated with strong sulphuric acid it burns with a characteristic
sugar smell .
2) Reducing : Monosaccharides which have free aldehydes or ketones reduce the Fehling's
or Bendict's reagent , thus the alkaline cupric ion ( blue colour ) present in this reagent
precipitated as reddish brown cupric oxide .
Cu2 + + aldose or ketone Cu2 O + Oxidation product
(Bendict Solution ) ( cuprous Oxide )
( blue ) ( reddish brown )
3) Oxidation : Oxidation of an aldose with bromine water , converts the aldehyde group to a
carboxyl group .
CHO ( aldehyde Group ) COOH ( carboxyl group )

( CHOH )4 Br2 + H2 O ( CH OH )4

CH2 OH CH2 OH
( Glucose ) ( Gluconic Acid )

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Qualitative Test
The qualitative tests are performed for identification of carbohydrates are following :

1) Fehling's Test :
In this test , 1-2 ml each of the Fehling's solution A and B are added to few drops of the test
solution and boiled for a few minutes . A Yellowish red colour appears that confirms the
presence of reducing sugar ( carbohydrate )

2) Tollen's test :
In this test , 2-3ml of Tollen's reagent is added to 2-3 ml of aqueous solution of
carbohydrate , and boiled in a water bath for 10 minutes . A shining silver mirror indicates
the presence of reducing carbohydrates .

Biological Roles Of carbohydrates

1) They are source of energy for living organism .
2) They are used as dietary fibers like cellulose .
3) they are used as flavouring and sweating agent .
4) They are stored in our body in the form of glycogen in liver and muscles and converted
into glucose to provide energy according to need .
5) They are important component of brain cells .
6) They are important component of DNA & RNA ( Deoxyribose and Ribose sugar )
7) They act as an anticoagulant in the form of Heparin .
8) They are major component of cartilage , tendon and bones.
9) They are used in clearance test in the form of Inulin .
10 ) They are used in the treatment of heart diseases in the form of Glycosides .

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Biochemistry Chapter 3

proteins
Definition
Proteins are organic compounds with high molecular weight , made up of long polypeptide
chain of different amino acids . or
proteins are hetropolymer compounds .

Amino Acids
these are building blocks of proteins , made up of carbon , hydrogen, amino group( NH2 )
and Carboxyl group ( COOH ), and R ( variable group ) .
H C is alpha carbon / chiral carbon .
H2 N C COOH
R ( side chain / variable group )

Different amino acids : about 300 amino acids are recognised , and actual number of amino
acids is unknown . some examples of different amino acids .
H
H2 N C COOH
H R1 ( Glycine )

H
H2 N C COOH
CH3 R2 (Alanine )
H
H2 N C COOH ( Serine )
CH2 -- OH R3

Polypeptide Chain :
When a large number of amino acids ( one hundred or more ) are linked together by
polypeptide bonds , is called polypeptide chain .
H H
H2 N C CO HN C COOH
R 1 ( Peptide bond) R2

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Classification of amino acids on the basis of chemical
nature
1 ) neutral amino acids : the amino acids have equal amount of amino and carboxylic group
are called neutral amino acids , examples : Glycine , alanine .
2) acidic amino acids : the amino acids have negative R group or chain are called acidic
amino acids , examples : aspartic acid , glutamic acids .
3) basic amino acids : the amino acids have positive R group or chain are called basic amino
acids , examples : arginine , lysine
4) Sulphur containing amino acids : methionine , cystine .
5) Aromatic amino acids : the amino acids have aromatic ring are called aromatic amino
acids . example phenylalanine .

Classification of amino acids on the basis of Nutritional

requirement
1) Essential Amino Acids : the amino acids are not synthesised in the body and are obtained
from dietary sources are called Essential Amino Acids .( VIPHALLMTT)
2) Non Essential Amino acids : the amino acids are synthesised in the body and are Non
Essential Amino acids .
Essential Amino Acids Non Essential Amino Acids
Valine Glycine
Isoleucine Tyrosine
Phenylalanine Proline
Histidine Cysteine
Arginine Aspartic acid
Leucine Alanine
Lysine Serine
Methionine Hydoxyproline
Tryptophan Cystine
Threonine Glutamic acid

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Biological Role of Amino Acids
1) they are building blocks of Proteins .
2) They are source of energy , during long term starvation , after consumption of stored
carbohydrates and fats .
3) Some of them are used as neurotransmitter.

Classification Of Proteins on the basis of Composition and

Solubility
Protein

Simple Conjugated Derived

globular Fibrous Nucleoprotein primary secondary

albumin collagens glycoprotein Proteins proteoses
Globulin elastins Mucoprotein peptons
Glutelins keratins Lipoprotein Proteans peptides
prolamines Phosphoprotein metaprotein
histones Chromoproteins
Globins metalloproteins
protamine

Simple Proteins :
* Simple proteins are made up of only amino acids , and they are classified as globular and
fibrous on the basis of axial ratio .
* Globular are soluble and fibrous are insoluble .

Conjugated Proteins :
* these proteins are made up of amino acids and non proteins substance ( prosthetic or
conjugated group ) .

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Derived proteins :
* These proteins are derived ( changed form ) from simple or conjugated proteins by
denaturation ( by heat, acid ,alkali, X-ray ) and Hydrolysis . It is in two types
1) Primary derived : In primary derived bonds are broken by denaturation except peptide
bonds .
2) Secondary derived : in secondary derived peptide bonds also broken by hydrolysis .

Structure Of Proteins ( Fore level of organization of protein structure ) .

1 ) Primary structure 2) secondary structure
3)tertiary structure 4) quarternary structure

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1) Primary Structure : The primary structure of protein indicate the arrangement of
different amino acids and they are joined by covalent peptide bond.
2) Secondary structure : it is the advanced form of primary structure, along with peptide
bond, hydrogen bond also found in secondary structure . Hydrogen bond is held between
amino groups and carboxyl oxygen of peptide bonds.
3) Tertiary structure : it is the more complex( folded ) form of secondary structure, tertiary
structure involves three dimensional folding of polypeptide chain, the polypeptide chain
are tightly folded into a complex form due to the following bonds :
hydrogen bonds, disulphide bonds , electrostatic or ionic bond and Hydrophobic bonds .
4) Quarternally structure: this is more advanced form of tertiary structure in this structure
two or more polypeptide chains interact with each other with non covalent interactions
and form a large complex of protein .
Qualitative test of protein
1) Ninhydrin test : amino acids and proteins a give Blue to violet colour with Ninhydrains
solution.
2) Biuret test: When biuret is treated with dilute copper sulphate in alkaline medium a
purple colour is obtained
3) Millon's test : when proteins are treated with Millon's reagent a white precipitate is
formed .
Biological role of Proteins
1)Protein as enzyme: proteins act as enzymes and play an important role in the regulation
of all the biological process in the body there are about 2000 different types of enzymes .
2)Protein as hormones: there are some proteins that function as hormone like growth
hormone , insulin.
3) Protein are associated with heredity characters: they help in transformation of genetic
information from one cell to another and from one generation to another
4) Protein as immune system: Immunoglobulins are made of proteins that make antibodies.
5) Transport : They transport specific types of molecule through the blood
6) Contraction : proteins like actin and myosin are responsible for the contraction of
muscles in living tissues .
7) Protein as receptor: proteins are present on the surface of cells and act as receptors
8) Protein as structural material: hairs, nails , wool are made up of keratin sulphate .
9) In bone : collagen proteins are found in cartilage .
10) in cell membrane : proteins are important part of membranes like plasma membrane ,
nuclear membrane.

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Diseases Related to Malnutrition( deficiency) of proteins
1) kwashiorkor : this disease occurs due to severe deficiency of dietary proteins .The
affected person have abnormally thin or week body parts , but have swollen ankles , feet
and belly due to fluid retention in them .

Symptoms
1) loss of weight .
2) growth and weigh gain stop .
3) Ankles , feet and belly swell .
4) Immune system is damage , and get infection easily .
5) The natural colour of skin and hair changes to a rust color .

Treatment
Kwashiorkor can be treated by increasing the proteins in diet .
2) Marasmus : This disease occurs due to severe deficiency of all macronurients ( proteins ,
carbohydrates , fats ) . Children are mostly suffer from this disease .

Symptoms
1) loss of weight .
2) growth and weigh gain stop .
3) The natural colour of skin and hair changes to a rust color .
4) chronic diarrhoea
5) respiratory infection

Treatment
Marasmus can be treated by increasing the proteins , carbohydrates , fats and other
essential nutrients in diet .

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Biochemistry chapter 4

Lipids
Definition : Liquids are Greasy , waxy or oily organic compounds with general formula Cn H2n
O2 , which are insoluble in water and soluble in organic solvent like ester , chloroform etc .

Classification Of Lipids
Lipids

Simple lipids Compound lipids Derived lipids

Triglyceride Phosphoglyceride steroids ( C29 , C28 )
Beeswax Kerasin terpenes ( mono, di , tri)
Carotenoids ( lycopene )

1) Simple lipids: these are esters of fatty acids which containing glycerol or alcohol and
carry no other substance .
2) Compound Lipids: these are Easter of fatty acids which containing an alcohol and other
groups also .
3) Derived Lipids : These are changed form of simple or compound lipids by hydrolysis.
Organization of lipids

Glycerol Fatty Acids

CH2 OH (C3 H8 O3 ) COOH --R

CH OH O

CH2 OH HO C R ( expended Form )

CH2 O C R

CH OH

CH2 OH ( Monoglyceride )

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O

CH2 O C R1

CH O C R 2

CH2 OH ( Diglyceride )

CH2 O C R1

CH O C R 2

CH2 O C R3 ( triglyceride )

Fatty Acids
Definition: Chemically, fatty acids are monocarboxylic acid with aliphatic carbonic chain .

Classification Of Fatty Acids On the basis of chemical nature

1) Saturated Fatty Acids : The fatty acids contain straight chain with single C--C band and
have no C C Double bonds are called saturated fatty acids . Examples Stearic acid ,
Palmitic Acid , Arachidic Acid .

COOH --R O H H H H H H H H H H H H H H H H H H
HO C C-- C--C--C --C--C--C--C--C--C--C--C--C--C--C--C--C--C --H
(Structure of fatty acid) H H H H H H H H H H H H H H H H H H
Omega Carbon
( Aliphatic Carbonic chain ) ( Omega 3 fatty acid )

2) Unsaturated Fatty Acids : The fatty acids have one or more C C

bonds in carbon chain are called unsaturated fatty acids . examples
Linoleic Acid , Linolenic Acid , Arachidonic acid

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Types of unsaturated fatty acids

a) monounsaturated fatty acids ( MUFA) : The fatty acid have one
double bond in carbon chain.
b) Polyunsaturated fatty acids ( PUFA ) : The fatty acid has two or more double
bond in carbon chain. example
O = Oleic Acid ( one double bond )
L = Linoleic Acid ( 2 double bond )
L = Linolenic Acid ( 3 double bond )
A = Arachidonic Acid ( 4 double bond )

Saturated fatty acids Unsaturated fatty acids

1 Fats Oils
2 Found in animal Found in Plants
3 Increase cholesterol level in blood Decrease cholesterol level in blood
4 Increase the risk of cardiovascular Lower the risk of cardiovascular
disease disease
5 Solid at room temperature Liquid at room temperature
6 Rancidity is high Rancidity is low
7 Butter, cream , meat etc. Almond oil , olive oil , coconut oil etc.

Nutritional requirement of fatty acid

1) Essential fatty acid: these are the fatty acids which are not produced in human body they
are must be taken through food (diet). examples Linoleic Acid , Linolenic
Acid , Arachidonic acid .
2) Non essential fatty acids : the fatty acids are produced in human body and no need to
take through foods ( diet ) . Examples Stearic acid , Palmitic Acid , Arachidic Acid .

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Structure And properties of triglyceride

Triglyceride are organic compounds in which three different fatty acids are linked to the
glycerol by Easter bond .

CH2 O C R1

CH O C R 2

CH2 O C R3 ( Triglyceride )

Properties of triglyceride
1) Non-polar: they are non polar( they does not have charge at the end)
2) In soluble in water: they are insoluble in water.
3) Soluble in organic substance: like Easter chloroform
4) Hydrophobic: they are hydrophobic in nature .
Structure and Functions of Cholesterol in the body

Cholesterol
Cholesterol is a type of lipids that found in all the cells of the body .

Structure Of Cholesterol

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Functions Of Cholesterol
1) It is a component of cell membrane .
2) It maintains the permeability of cell membrane .
3) It helps in bile production ..
4) It helps in the formation of vitamin D .
5) It metabolize ( break down ) fat soluble vitamins .
6) It helps in the formation of male and female sex hormones .
7) it insulate the nerve fiber .

Lipoproteins
Lipoprotein : Lipoproteins are combination of lipid and protein or we can say conjugated
protein .

Types of Lipoproteins
On the basis of electrophoresis separation , five types of lipoproteins are found in human
plasma .
1) Chylomicrons : This types of lipoproteins are synthesized in the intestine and have the
highest lipid content (98% ) and lowest protein content (2%) , and have the lowest density
and are biggest in size . (have 4% of cholesterol of 98% of lipids)
2) Very low density lipoproteins (VLDL): these proteins are produced in the liver and
intestine , and have 10% of proteins and 90% of lipids . (have 24% of cholesterol of 90% of
lipids) .
3) Low Density lipoproteins( LDL ) (Bad Cholesterol ) : these lipoproteins are produced in
the blood stream from VLDL and transport cholesterol from liver to the rest of the body,
and have 20% of proteins and 80% of lipids . (have 56% of cholesterol of 80% of lipids)
4) High density Lipoproteins ( HDL ) (good cholesterol) : These lipoproteins are produced in
the liver and transport cholesterol from peripheral tissues (skeletal muscle , adipose ) to
liver , and they have 40% of proteins and 60% of lipids . (have 40% of cholesterol of 60% of
lipids).

Function of lipoproteins In the body

1) Absorption and transportation of lipids in small intestine .
2) transportation of lipids from liver to tissues .
3) transportation of lipids from tissues to liver .

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Qualitative Test of Lipids
1) Grease spot Test : In this test one drop of oil and a drop of water are put on a paper . at
first , both areas look wet but after some time water is evaporates and the spot dries
whereas lipid spot remains visible .
2) Solubility test : In this test we add some drops of sample of lipid to organic substance
and water and observe . it will miscible with organic substances like chloroform , ether and
immiscible with water .

3) Emulsification test

4) Saponification Test

Function Of lipids
1) Source of energy : it is a best source of energy it provide energy more that
carbohydrates and proteins .
2) Cell membrane : they are important component of the cell membrane (phosphoric)
3)Membrane permeability: they regulate membrane permeability .
4) Vitamins: they store fat soluble vitamins (KEDA)
5) Enzyme: they are components of various types of enzymes.
6) Thermal insulation: fat deposited in the subcutaneous layer provide insulation and
protection from cold .
7) Transport : lipoproteins proteins transport cholesterol and triglyceride from their origin
to site of use .
8) Electrical insulation : they act as electrical insulator to the nerve fibres in the form of
myelin sheath .
9) Storage : they Store meaning of compounds .
10) Signalling : they act as signaling molecule .

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Biochemistry Chapter 5
Nucleic Acid

Definition Of Nucleic Acids :

Nucleic Acids are intracellular organic substances , made up of polymerization
of Nucleotide . They store genetic information and enable protein synthesis
or production . composition of Nucleotide

Phosphate group pentose sugar nitrogenous base

( deoxyribose , ribose )

Nitrogenous Bases
Purines : they contains two, carbo - nitrogen rings one of them is six
membered and other is 5 membered . example Adenine , Guanine .
Pyrimidine : It is a 6 membered carbo-nitrogen ring , at the 1st and 3rd
position it contains N . example Cytosine , thymine , Uracil .

Component Of Nucleoside and Nucleotide

Nucleoside : it is made up with the combination of pentose sugar and
nitrogenous base by glycosidic bond .

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Nucleotide : nucleotides are organic substances made of nucleoside and
phosphate .
or nucleotides are organic substances made of pentose sugar , nitrogenous
base and
phosphate group.

( Phosphoester bond) ( glycosidic bond )

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Structure Of DNA ( Waston and Crick model ) and Function .

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Function Of DNA
1) Genetic Information : It transfer genetic information from mother to the
child .
2) Replication : It makes its copy during cell division.
3) Transcription : When RNA is formed from DNA it is called transcription .
4) DNA finger print : It help to determine the identity of suspected person on
the basis of nucleotide sequences .

Structure and Function of RNA

Function Of RNA
1) mRNA : It takes genetic massages from DNA and help in protein synthesis .
2) t-RNA : It transfer amino acids to the codes information of mRNA .
3) r-RNA ( ribose RNA ) : It play an important role in protein synthesis .

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Biochemistry Chapter 5
Enzymes

Definition : The specific type of proteins that c atalyse ( increase the rate of )
chemical reactions in living cells are called enzymes . or
Enzymes are catalysts(a thing that causes changes) that catalyse the chemical
reaction .

Properties Of Enzymes
1) Enzymes increase the rate of reaction .
2) they are always specific for their substrate .
3) Enzymes are thermolabile ( means heat sensitive , they can be denatured
by heat )
4) Enzymes are amphoteric ( they have positive and negative charge due to
basic and acidic functional groups ) .
5) they are unstable and their activity can be decreased or enzymes can be
destroyed by variety of conditions like temperature , wide changes in pH .
6) they are proteins with high molecular weight ranging between 10000 to
millions .
7) All enzymes are protein s , except the group of catalytic of RNA .

IUB ( International Union of Biochemistry )and MB (molecular Biology)

Classification
1) Oxidoreductases: these type of enzymes involve in oxidation( loss of
electron) reduction( gain of electron ) example s dehydrogenases , oxidases .

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2) Transferases : these type of enzymes involve in transferring of chemical
group such as alcohol, carboxyl , sulphate , phosphate, aldehyde, ketone .
example transaminases , kinases .
3) Hydrolyses : These type of enzymes involve in breaking of different bonds
.Examples Lipase , peptidase , glycosidase .
4) Lyases : These type of enzymes involve addition or removal of the
elements of water (hydrogen, oxygen), ammonia (nitrogen, hydrogen), or
carbon dioxide (carbon, oxygen) at double bonds. For example,
decarboxylases remove carbon dioxide from amino acids and dehydrases
remove water.

5) Isomerases : These type of enzymes are involve in isomerization reaction

( Isomerization is a process in which one isomer is formed from another. An
isomer is any of the two or more forms of a molecule with the same chemical
formula but with a different stereochemical arrangement of the atoms).

6) Ligases : these type of enzymes involve in biding or joining of two molecules

together.

e.g. Synthetases , carboxylases .

Factors Affecting Enzymes Activity

1) Concentration of Substrate .
2) Concentration of Enzymes .
3) Effect of temperature .
4) Effect of pH .
5) Effect of UV rays .
6) Effect Of Inhibitors
7) effect of Activators .
1) Concentration of Substrate .
If the concentration of substrate is increased by keeping the concentration of
enzyme constant , initially rate of reaction increases , but when all enzymes
are occupied by substrate , further increase in concentration of subs trate
does not increases the rate of reaction .
2) Concentration of Enzymes :
If the concentration of enzymes is increased by keeping the concentration of
substrate constant , initially rate of reaction increases , but when all

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substrates are occupied by Enzymes , further increase in concentration of
enzymes does not increases the rate of reaction .
3) Effect of temperature .
The rate of enzyme reaction increases initially as temperature increase . But
this increase is obtained till optimum Temperature ( 35 - 38 ° C ) . further
increase in temperature may denature the Enzymes and rate of action will
decrease .
4) Effect of pH .
There is a range of optimum pH in which enzymes maximum Activity , this
range is generally from pH 6.4 - pH 7.5 . Exceptionally there are some
enzymes for them optimum pH is Highly acidic or slightly acidic or alkaline
.They are given Below .

Enzymes Optimum pH
1) Acid Phosphatase 5.5
2) Pepsin 2. 0
3) Amylase 6 to 7
4) Lipase 7.0
5) Papain 5 to 6
6) Trypsin 6 to 8.5
7) alkaline Phosphatase 9.2

5) Effect of UV rays .
UV rays denature the enzymes so enzyme activity decrease .

6) Effect Of Inhibitors
Inhibitors prevent the action of enzymes , so they decrease the rate of
reaction .

7) effect of Activators .
Activators increase the action of enzymes , so they increase the rate of
reaction.
Mechanism of Action of Enzymes
Step 1 : - The enzyme attracts the substrate to its active site and becomes
Enzyme - Substrate complex .
E+S ES complex
Step 2 :- a process called catalysis occurs
Step 3 :- Now The enzyme release the Substrate and now substrate is called
Product (p) .
ES E+P

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There are two models which explained the Me chanism of action of enzyme :
1) Lock and Key Hypothesis ( Emil Fisher )
2) Induced fit Hypothesis ( Koshland ) .
Enzyme Inhibitors
The agents which prevent or decrease the action of Enzymes are called
Enzyme inhibitors :
Types of Enzyme Inhibitors
1) Reversible / competitive Inhibitors : This type of Inhibitors have structural
similarity with substrate , and bind at the active site of enzymes and inhibit the action of
enzyme .
If the concentration of Substrate increase then the inhibition is revers ed .
2) Non - Competitive or Irreversible Inhibitors : this type of Inhibitors bind
with functional group other than the active site , and inhibit the action of
enzymes for ever .
If the concentration of Substrate increase then the inhibition does not
reverse

Therapeutic and Pharmaceutical Importance

Therapeutic Importance :
1) Galactosidase is used for treatment of Lactose intolerance in children .
2) L- asparaginase is used to treat Cancer( Leukaemia )
3) Pepsin , Lipase , amylase , Peptidase ar e used to treat gastrointestinal
digestive disorder .
4) trypsin ,is used to treat Inflammation .
5) Nattokinase is used to increase production of Plasma .

6 ) streptokinase are used to treat thrombosis .

7) collagenase is used to treat skin ulcer .

8) Uricase is used to treat Gout .

9) Lysozyme is used as antibiotic .

10) urokinase is used to dissolve blood clots .

Pharmaceutical Importance enzymes ( used in manufacturing of drugs )
1) Glucose isomerase is used for production of High fructose syrup .
2) Gama glutamyl Transpeptidase is used for diagnosis of alcohol abuse and
infective hepatitis .

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3) Penicillin acylase is used in production of penicillin .
4) Lactase Dehydrogenase is used for diagnosis of infective hepatitis
,Leukaemia
5) Amylase is used for diagnosis of Pancreatitis .( inflammation of pancreas ,
in which serum amylase level increase ) .
6) Gama glutamyl Transpeptidase is used for diagnosis of alcohol abuse and
infective hepatitis .

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Biochemistry Chapter 7
Vitamins

Definition
Vitamins are those Organic Substances that maintain the normal structure and
function of cells .
Vitamins are not synthesised by human body , that is why they are taken in
food from external source ( plants , animals ) .
Classification
1) Fat soluble Vitamins : Vitamin A, D,E , K ( KEDA) are fat soluble vitamins .
They are hydrophobic in nature and are stored in liver .
These are not excreted out from body , so excessive intake can cause toxic
effect which is called Hype rvitaminosis .
2) Water soluble Vitamins : Vitamin B Complex and Vitamin C are water
soluble Vitamins .
These are not stored in the body , therefore they are required daily in small
quantity .

Scientific name of Vitamins

Fat soluble Water soluble
Vit. A ( Retinol ) B 1 ( Thiamine )
VIt. D ( Calciferol ) B 2 ( Riboflavin )
Vit. E ( TocoPherol ) B 3 ( Niacin )
Vit . K ( Phylloquinone ) B 5 ( Pantothenic Acid )
B 6 ( Pyridoxine )
B 7 ( Biotin )
B9 ( Folic acid )
B 1 2 ( Cobalamin , Cyanocobalamin )

Vitamin A ( retinol )
Functions
1) It is necessary for normal Vision .
2) It is important for body growth .
3) It helps in growth of bones and Neurological System .
4) It keeps the skin , Kidneys , and other org ans perfect .

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Deficiency Diseases
1) Night Blindness 2) xErophthelmia ( dryness of eye )
3) Keratomalacia ( ulcernation of cornea ) 4) Dryness of skin .
Daily Dietary requirement
600 mcg /d
Sources :
Green Vegetables , fruits , cereals , liver , milk , Butter , and egg yolk .
overuse ( Hypervitaminosis A )
Overuse cases Headache , nausea , Vomiting , tiredness etc.
Vitamin D ( Calceferol )
Functions
1) It increases the absorption of Calcium and Phosphorus from Intestine
2) It promotes the normal growth and development of bones .
3) It keeps bones healthy .
4) It promotes the normal growth of the body .
Deficiency Diseases
1) Rickets in children 2) Osteomalacia
Daily Dietary requirement
100 units / day ( 2.5 mcg) . an d lactating mother 400 units /d
Sources :
Cod liver oil , fish liver Oil , egg yolk , animal liver .
Overuse
Over intake of Vitamin D ( 100 mcg or 1000 IU ) on regular basis can cause
Kidney stone , anorexia ( loss of appetite ) , Polyuria , Constipa tion , excessive
thirst , weight loss etc.
Vitamin E ( Tocopherol )
Functions
1) it is helpful for healthy reproductive system .
2) it is an good antioxidant .
3) it prevents liver cells from toxicity .
4) it protect nerve fibres from demylination
5) it prevents rancidity .

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Deficiency diseases
1) Tooth discoloration 2) weakness of RBC
3) Thrombosytosis and oedem 4) Sterility in male , death of fetus in
uterus in female
Daily Dietary requirement
10 mg/day and for pregnant and lact ating woman 35mg /d .
Sources :
Meat , liver , fish , chicken , vegetable oils , green leafy vegetables ,egg yolk .
Vitamin K ( phylloquinone )
Functions
1) it increases the activity of blood clotting factors .
2) it helps the liver to manufacture th e enzyme that catalyses the formation
of prothrombin .
Deficiency diseases
1) Delay in blood clotting 2) improper functioning of liver .
Daily Dietary requirement
140-200mg/d
Sources
green leafy vegetables( Spinach , Cabbage etc.) , Soya beans , wheat germ .
Vitamin B 1 ( Thiamine )
Functions
1) It is necessary for maintaining good appetite .
2) It helps in utilizing the carbohydrates in body .
3) it is needed for brain metabolism .
Deficiency diseases
1) Beri-Beri ( nerve system problems , oedema , weakness , headache ,
Insomnia , GIT disorders .)
2) Anorexia ( loss of appetite )
Daily Dietary requirement
1.5mg /d
Sources :
rice bran , wheat bran , whole grains , nuts , pulses , beam , yeast , liver , eggs
, fish , meat , and milk .

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Vitamin B 2 ( Riboflavin)
Functions
1) It helps the regulation of some carbohydrate - metabolising hormones.
2) In presence of light , it stimulate the optic nerve .
Deficiency diseases
1) injury of lips 2) Cracks at the mouth' s angle )
3) Eczema on face ( red and dry skin and itch ) 4) Hair loss
5) red eye reproductive problems 6) cataract
Daily Dietary requirement
0.5mg/d
Sources
Milk , liver , eggs yolk , sprouts cereals .
Vitamin B 3 ( Niacin )
Functions
1) It perform metabolic function
2) It converts carbohydrates into glucose .
3) it keeps nervous system working properly .
Deficiency diseases
1) dermatitis ( Itchy inflammation of skin )
2) Diarrhoea
3) Demantia ( memory loss , neuronal disorder )
Daily Dietary requirement
6 mg/d
Sources
Liver , fish , bean , and peanuts .
Vitamin B 5 ( Pantothenic Acid )
Functions
1) It is essential for growth of infants and children .
2) It is required for Breakdown of fasts and carbohydrates for energy .
3) It is required for formation of RBCs .
Deficiency diseases
1) Anaemia 2) growth problem 3) Liver problems
4) nausea 5) Vomiting 6) Fatty liver .

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Daily Dietary requirement
6mg/d
Sources
Honey , cereals , nuts , eggs , liver ,fish , milk , meat , bran .
Vitamin B 6 ( Pyridoxine )
Functions
1) It is needed for Infant growth .
2) It is required for formation of dopamine , serotonin etc.
3) Formation of RBCs and WBCs
Deficiency diseases
its deficiency rarely occurs
1) depression 2) confusion 3) decrease immune function . 4 anaemia
Daily Dietary requirement
1.6-2 .0mg/d
Sources
Milk , liver , eggs yolk , meat , leafy vegetables etc.

Vitamin B 7 ( Biotin )
Functions
1) It helps in metabolism of carbohydrates , lipids , pr oteins and produces
energy .
Deficiency diseases
1) alopecia ( hair loss ) 2) hair graying .
Daily Dietary requirement
100-300mcg /d
Sources
Honey , Milk , liver , eggs yolk , meat , fish etc.
Vitamin B 9 ( Folic acid )
Functions
1) it help in Protein metabolism ,.
2) Controls Homocysteine level
3) it break Homocysteine amino acid .
Deficiency diseases
1) affect DNA Synthesis
2) Megaloblastic anaemia . ( abnormal large Red Blood cells ) .
Daily Dietary requirement
300-400 mcg /d
Sources

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leafy vegetables liver , eggs etc .
Vitamin 1 2 ( Cobalamine )
Functions
1) it is required for formation of genetic materials .
2) For formation and maturation of RBCs
3) it is needed for maintenance of nervous system .
Deficiency diseases
1) neurological disorder 2) Infertility 3) Demyelination
4) Megaloblastic anaemia .
Daily Dietary requirement
5mcg /D
Sources
liver, milk, moulds ,eggs .

Vitamin C ( ascorbic acid )

Functions
1) It is necessary for Iron absorpti on .
2) It provides immunity against viruses and cold .
3) It keeps gums and capillaries healthy .
4) It helps in wound Healing .
Deficiency diseases
1) Scurvy disease
a) bleeding from gums and decaying teeth .
b) loosing teeth
c) haemorrhage due to weakness of capillaries .
d) delay in wound healing .
Daily Dietary requirement
30mg/D for infants and 70mg/D for adults .
Sources
Citrus Fruits , grapes , apple , papaya , guava , vegetables etc

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Co-Enzymes

Definition
Co-Enzymes are a non protein Compound that bind with Enzymes and increase
the function of Enzymes .
All the water soluble vitamins and two of the fat soluble vitamins A and K act
as Co-enzyme or Cofactors .

Co-enzyme Forms of Vitamins

Water soluble Coenzyme Form

B 1 ( Thiamine ) Thiamine Pyrophosphate ( TPP )
B 2 ( Riboflavin ) Flavin Mono Neucleotide ( FMN) , Flavin
Adenine Dineucleotide ( FAD )
B 3 ( Niacin ) Nicotinamide Adenine Dineucleotide ( NAD )
Nicotinamide Adenine Dineucleotide
Phosphate
B 5 (Pantothenic Acid )
Coenzyme A
B 6 ( Pyridoxine )
Pyridoxal Phosphate
B 7 ( Biotin )
Biotine Carboxyl - carrier Protein
B9 ( Folic acid )
Tetrahydrofolic Acid
B 1 2 ( Cobalamin ,
Co -enzyme B 1 2

Chemical Nature of Vitamins

Vitamin A
It is insoluble In water , soluble in ethanol and freely soluble in organic solvent like fats and
oils .
It is an antioxidant .
Low sensitive light , heat and Iodine
Retinoids occurs in these three forms
a) Alcoholic Retinol b) aldehyde retinol c) Acid retinoic Acid .
Vitamin D

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It is insoluble in water , moderately soluble in fats ,oils and ethanol , freely soluble in aceton
, ether , and petroleum ether .
It is sensitive to oxygen , light and iodine .
heating , or mild acidity can converts it to the inactive forms .
Vitamins E
It is insoluble in water ,and soluble in fats and oils .
they are good antioxidants .
they are resistant to acid .
they are stable in air .
it is acidic in nature ( acetic acid )
it is oil at room temperature .
Vitamins K
Insoluble in water , and slightly soluble in ethanol and freely soluble in ether , chloroform ,
fats and oils .
It is sensitive to light .
Vitamin B1
It is highly soluble in water moderately soluble in methanol and glycerol , and insoluble fats
and oils .
free thiamine is unstable , that is why hydrochloride and mononitrate forms are used .
it is colourless crystal
Vitamins B2
It is moderately soluble in water
insoluble in fats and oils
It is light sensitive , so should be handle in dark ,
It can not be extracted with usual organic solvent , it is extracted with chloroform .
Vitamins B3
It is of amphoteric nature and forms salts with acids as well as with bases .
It is soluble in water .and insoluble in fats .
It is stable in dry form
Vitamin B5
It is yellow viscous oil .
It is soluble in water and ether , and in soluble in organic solvents like fats acids .

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It is unstable to heat and acidic or alkaline condition .
Vitamins B6
It is colourless crystal at room temperature .
It is highly soluble in water and insoluble in chloroform and fat .
it is stable in solution
Vitamins B7
It is white crystalline substance .
It is stable to air , heat and light in dry form .
It degraded under strongly acidic or basic condition .
It is soluble in water , and insoluble in fat .
Vitamin B 9
it is an organic yellow crystalline substance
soluble in water and insoluble in fats .
unstable to light .
Vitamin B 12
it is red , red -orange or yellow crystalline substance .
Soluble in water and insoluble in fats .
stable to heat
It is more stable in presence of ascorbic acid .
Vitamin C
* It is soluble in water and insoluble in fats .
* In aqueous solution , Dehyroascorbic acid is unstable .
* Dehydroascorbic acid reacts with several amino acids .

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Biochemistry and clinical Pathology Chapter 9
Minerals

Definition
Minerals are the elements which are necessary for different types of Physiological functions
and many of biochemical process .
Types
1) Macro-Minerals or Essential or Major Minerals : The minerals are required more than
100mg per day as a daily requirement are called Macro-minerals such as : Sodium ,
Potassium , Calcium , chloride , Magnesium , Phosphorus .
2) Micro-Minerals or Trace Minerals : The minerals are required less than 100mg per day as
a daily requirement are called Macro-minerals such as : Iodine , Fluorine , Copper , Cobalt ,
Manganese , iron , zinc, Selerium , Sulphate , Arsenic , Vanadium .

Functions of Minerals
They are required for :
1) for transport of Oxygen ( Fe)
2) For maintenance of tissues and bones . ( Ca++ )
3) For working of nervous system . ( Ca++ )
4) For muscle contraction . ( Ca++ )
5) For blood coagulation ( Ca++ )
6) For Cardiac activity ( K+ )
7) For acid-base balance ( Na+ )
8) For thyroid Hormone synthesis ( Iodine )
9) for formation of sperms ( Zn )
10) for maintenance of Osmotic pressure of blood ( Na+ , K+ , Cl - )
Macro-Minerals
1) Calcium: Calcium is a main component of bone and teeth , calcium is taken in diet in the
form of Calcium Phosphate and calciumcarbonate . It is the most abundant mineral in
human body ( 1200 g approximately).

Functions
 For formation and development of bone and teeth .
 Calcium is necessary for bone calcification ( it is a process in which calcium builds up
in the bones and teeth and make them harden .

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 It is helps in the blood coagulation , helps in conversion of prothrombin into
thrombin .
 It helps the body in muscles contraction .
 It activates enzymes .
 It acts as an intracellular Messenger of different Hormones .
 It increase the permeability of Plasma membrane .
 It plays an important role in action Potential .
Deficiency Diseases
1) Hypocalcemia ( Low blood calcium level )
2) Osteoporosis ( weak and fragile )
3) Tetany ( Involuntary muscles contraction due to lack of calcium in blood )
4)Rekets ( in children )
5) osteomlacia ( In adult )
Recommended Dietary Requirement
500m g /day for a healthy adult , 1.5 - 1.0 g /d for Pregnant women .
2) Phophorus : It is also a major component of bones and used for Energy production .
Functions
 Formation and development of bone and teeth .
 For formation of nucleic acids and Phospholipids .
 for formation of Co-enzymes like NADP,
 For formation of ATP , ADP, AMP .
Deficiency Diseases
1) hypophosphatemia .
2) Rikets
3) Osteomlacia
Recommended Dietary Requirement
500-700 mg /d for a healthy adult .
3) Sodium : It is the chief cation of the extra cellular compartment , around 50mmol/kg
sodium is found in a healthy man .
Function
 It involves in maintaining body fluid through osmosis .
 It retains water in the body .
 it helps in excitability of nerves .
 it helps in acid - base balance .
 it helps in maintaining viscosity of blood .

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 it maintains electrolyte balance .
Deficiency Diseases
1) Hyponatremia
Recommended Dietary Requirement
5-6 g / for a healthy adult
4) Potassium : It is major intracellular Cation , and performs following functions :
Functions :
 For acid-base balance
 It helps in cardiac muscles function .
 For neuromuscular function activity .
Deficiency diseases
Hypokalemia

Recommended Dietary Requirement

2-2.4 g for a healthy adult .
5) Chloride : It is major anoin in the body . Around 125mmol/l chloride is found in
cerebrospinal fluid ( CSF) . about 80 g chloride ions found in a healthy adult .

functions
 It involves in acid-base balancing .
 it helps in secretion of HCL in stomach .
 it helps in maintaining fluid balance by osmosis .
Deficiency Disease
Hypochloremia

Recommended Dietary Requirement

5-10 g for a normal adult
6) Magnesium
Functions
 It helps in activation of enzymes .
 it is required for DNA replication , transcription , and translations .
Deficiency diseases
Fatigue , Weakness , appetite loss , nausea and vomiting .
Daily requirement
350 mg/d for a adult man .
Micro-Minerals
1) Iron : It very important for formation of haemoglobin , around 3-5 g oiron is found in a
healthy adult .

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Functions
 It required For formation of Hb /RBCs
 It required For formation myoglobin .( it store oxygen in muscle cells )
 it required for synthesis of DNA .
 It transport O2 Co2 .

Deficiency diseases
Iron deficiency anemia

Recommended Dietary Requirement

15-20 mg /d for a healthy adult
2) Iodine :
functions
 It required For synthesis of thyroid Hormone
 It required for growth and development of body .

Deficiency diseases
Goiter , Hypothyrodism , Cretinism ( stopped physical and mental growth ) .

Recommended Dietary Requirement

120- 140 mcg

3) Copper

functions
 It involves in haemoglobin synthesis
 it required for melanin formation .

Deficiency diseases
Hypochromic ,

Recommended Dietary Requirement

about 900 mcg /d for a normal adult .
4) fluorine
functions
Tooth development , Dental health , Bone development ( it increase calcium and
phosphate reaction ) .
Deficiency diseases
Dental caries
Recommended Dietary Requirement
2-4 mg /d for adults

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5) Zinc
functions
It requires for enzyme formation
it requires for sperm formation .
it helps in production of DNA and proteins .
It helps in wound healing

Recommended Dietary Requirement

8 - 11 mg for adults

Deficiency diseases
Diarrhoea , Skin disorders , obesity , diabetes mellitus. Delay in wound healing

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Biochemistry chapter 10
Water and Electrolytes
Introduction
Water is the most important component of the human body .It is necessary
for normal function of the body. Chemically water is an inorganic compound ,
its molecular formula is H 2 O . It forms approximately 55-660% mass of an
average adult's Body .
The total water content decrease in older people due to reduced muscles
mass .
Distribution of Water in the Body
Total Body Weight

Total Body water 42 L ( 60%) of TBW Other things ( 40%)

Intracellular Fluid ( ICF) 28 L 65% of T.B.water Extracellular Fluid (ECF) 14 L ( 35 % ) of TBW

Intestinal Fluid 10 .5 L ( 25% tbw.) Plasma Fluid 3.5 L ( 8 %)

Distribution Of water in different body organs

1) Brain and heart are composed of 73 % of water
2) Lungs are composed of 83 % of water
3) Skin are composed of 64 % of water
4) Muscles and kidneys are composed of 79 % of water
Function of Water in the body
There are a lots of function of water in the body s ome them are following :
 It maintains body temperature .
 It provides lubrication in joints and protects the bones .
 It makes the blood plasma thin and helps in proper flowing to tissues .
 It helps in excretion of wastes from body .
 It helps in intracellular transportation .
 it prevents dryness of skin .
 it helps in occurrence of many enzymatic reaction .
 it helps in formation of saliva .
 it prevents constipation .
 It helps in maintenance of blood pressure .
 It helps kidney to perform properly .
 It helps in the maintenance of pH of the body .

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Water turnover and Balance
The water turnover is defined as " the amount of water entering in the body
in comparison to the amount of water loss in a specific time periods " . and
having water in appropriate amount is called Water balance .
Normal Water level ( 2500ml/d)
In the normal resting state , input of water through ingested fluid is
approximately ( 1200ml/d) . from ingested food ( 1000ml/d) and from aerobic
respiration 300ml/d . Totaling 2500ml /d ( 2.5 L ) .
The measurement of Hydration
Plasma osmolality , urine osmolality are widely used markers for measuring
the hydration level of the body .
Osmolality : the concentration of particles dissolved in fluid .
Electrolytics
 Electrolytes are minerals which dissolved in water and release ions .
 They are found in body fluids also like sodium , Chloride , potassium ,
and maintain osmotic pressure .
 They maintain amount of water in the body , particularly Sodium
maintains water balance through Osmosis .
Electrolyte Balance
If electrolytes are present in proper amount as body needs it is called
electrolyte balance .
Electrolyte Composition of Body Fluids
Cations (mmol/l) Plasma Intestine Intracellular
Na 142 139 14
K 4.2 4.0 140
Ca 1.3 1.2 0
Mg 0.8 0.7 20
Anions (mmol/l)
Cl 108 108 4.0
HCO3 24.0 28.3 10
Protein 1.2 0.2 4.0
HPO4 2.0 2.0 11

Dehydration
Dehydration is a condition in which the body 's Intravascular , Intestine and
intracellular fluid levels decrease , and cause defi ciency of water and
Electrolytes .

Causes
1) heavy loss of gatrointestinal fluids due to vomiting , Diarrhoea or by any
other reason .
2) heavy blood loss .
3) storing of fluids in an area of body where it does not performs normal
physiological process .

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Oral Rehydration Therapy
Oral rehydration therapy is a very simple and effective therapy to treat
dehydration . In this therapy ORS is used and it can be made at home using
salt and sugars in clear water .
formula to make ORS at home .
Clean water 1 liter
Sugar 6 level teaspoons ( 30 g)
salt half level teaspoon ( 2.5 g)

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Biochemistry and clinical Pathology Chapter 11

Introduction to Biotechnology
Biotechnology is a branch of science in which using biology (living cell or bacteria or any
part of them) and technology or scientific process , a new product is developed to
improve human health and environment .
Biotechnology is also called "biotech ".
there are many subfields of biotechnology ,the main subfields are these :
1) Medical ( red ) biotechnology
2) Agriculture ( green) biotechnology
3) Industrial (white) biotechnology
4) Marine (blue) biotechnology
1) Medical biotechnology : It is used for medicinal purpose for example gene therapy is
used to treat genetic or acquired disease like cancer this therapy utilizes normal genes for
replacing the defective gene .
2) Agriculture biotechnology : this is related to agriculture product and processes for
example one or two genes are combined together and developed a new variety of crop for
increasing the yield
3) Industrial biotechnology: this is related to industrial process for example replacing genes
of a microorganism a new organism id developed to produce a useful chemical .
4) Marine biotechnology : this field involve in Marine resources to develop a novel
pharmaceutical , drugs , chemicals products , enzymes or other industrial product .

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Chapter 12

Organ Function Test

Function of kidney
1) Regulation of blood volume : they regulate blood volume by conserving the water or by
excreting water in urine
2) Regulation of blood pressure: They regulate BP by regulating Blood volume .
3) Regulation of renin enzyme : renin is an enzyme which is produced by kidney and helps
in the formation of angiotensin , which causes vasoconstriction and increased blood
pressure .
4) Regulation of blood pH : they regulate the pH of blood by excreting h+
ions into the urine and conserve HCO3-- ions , (it is an important buffer ions in the blood)
5) Regulation of blood ionic composition : the regulate the concentration of some ions like
Na+ , K+ , Ca++ , Cl-- etc.

6) Production of hormone : they produce Calcitriol (the active form of vitamin D

)which regulate the calcium homeostatasis ( the maintenance of constant concentration of
calcium ion in the extracellular fluid )

7) Stimulation of RBC production : They produce erythropoietin Hormone which

stimulate RBC production .
8) Regulation of blood glucose level : they take part in maintenance of blood glucose level
as they increase blood glucose level by gluconeogenesis process
9) Excretion of Wastes from body : they excrete out waste materials from body like
ammonia , urea , bilirubin , creatinine , uric acid , drug and toxins.
Renal Function Tests (KFT Test)
Kidney or renal function tests are performed using urine ( and some test are
performed using blood like urea clearance test) , and the presence of
different substances in abnormal amount in urine or in blood indicates the
kidney problems .

Physical examination of urine

Colour
Odour
pH ( 4.5-8 )
Turbidity

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Chemical examination of Urine
1 ) Proteins : Globulin less than 5 mg/dl and albuline less than 30 mg/dl .
2) Abnormal Glucose level : normal is 0-0.8 mmol /L .
3) Ketone Bodies : Under normal condition , < 1 mg of Ketone Bodies is
excreted in 24 hours .
4) Blood : blood in urine indicates Sore of kidneys , UTI and nephritis .
5) Urine Osmolality Test : Urine osmolality is the number of dissolved
particles in urine( creatinine , urea, potassium , sodium etc.) . Normal Range :
500-850 mOsm/kg .

Significance :
Increased level indicates Kidney probl ems and congestive heart failure . and
cause dehydration .
6) Urine Concentration Test : kidneys maintain the osmolarity of body fluid (
290-300 mOsmol/L) , and excrete urine with average 500 -850 mOsm/l
normally

significance :
High concentrated urine indicates Kidney problems , dehydartion , heart
failure .

some blood sample test are performed to Evaluate kidney function .

1) Blood Urea nitrogen test ( BUN ) : Urea is obtained as a by-Product of
protein metabolism . It is formed in liver and excreted ou t in urine through
filtration of blood by kidneys.
Normal range 10-12mg /dl
significancec : increased level of Blood urea nitrogen indicates kidney problems
.
2)Creatinine Test : Creatinine is a waste product of body metabolism which
present in blood excreted out by Kidneys .
Normal Range Creatinine in blood
1-2mg/dl or 0.6-1.2 mg/dl

Significance
Increased level in blood and low level in urine indicates kidneys dysfunction .
Urine Creatinine normal Range
for adult man : 0.74 - 1.35 mg/dl
For adult woman : 0.59 - 1.04 mg/dl

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3) Creatinine Clearance Test
This test shows the ability of kidneys to clear( Excrete out) creatinine from
blood through urine. In this test 24 hours collected urine sample is taken.
along this test blood creatinine level also examined .
Normal Range of Creatinine clearance /min in urine
in adult men : 90-139 ml/min
In adult women : 80- 125 ml / min

Significance :
Lower than normal creatinine excretion indicates the kidney problems .

4) Urea Clearance test

This test is also performed to check the kidneys functions . In this test using
blood , the amount of urea in blood Checked , and along with two urine
sample are collected with a gap of one hour to determine the amount of urea
filtered by the kidneys into urine .

Normal Range of urea clearance

12-20 gm/24 hours

Significance
Low level of urea than normal range indicates kidney problems , Protein deficiency in diet .
High level than normal range indicates excessive protein metabolism , or too much protein
intake in diet .

Function of liver
1) Metabolism of carbohydrate : liver Plays an important role in the
metabolism of carbohydrates and release glucose into blood , in case blood
glucose level is high liver convert glucose into glycogen and store it .
2) Metabolism of protein and lipids : In case glycogen is not enough to fulfill
the body requirement of glucose , lever makes glucose from protein and fat
which is called gluconiogenesis .
3) Detoxification : Liver detoxifies the toxic substance like al cohol drug and
steroid hormone and prevents other tissue from damage .
4) Storage : liver stores glycogen , certain vitamins (fat soluble ) and
minerals (iron and copper)

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5) Phagocytosis : the aged RBCs , WBC s and some bacteria undergo
phagocytosis by kuffer cells of liver and destroyed .
6) Formation of urea : the ammonia is obtained during metabolism of protein
which is highly toxic is converted into urea which is less toxic.
7) Formation of RBC in foetal life .
8) Destruction of aged RBC and f ormation of bile pigment .
9) Formation of plasma protein : like albumin , globulin prothrombin and
fibrinogen .
10) Formation of heparin : it is a natural anticoagulant present in the blood .

Liver Function Tests (LFT Test)

Liver function test is performed to estimate the dysfunctions of liver several
testes are performed under liver function test .Like Serum Bilirubin , serum
(Plasma) proteins , Alkaline Phosphatase ( ALP) , Serum Glutamic Oxaloacetic
transaminase (SGOT) or Aspartate transaminase (AST), Serum Glutamate
Pyruvate Transaminase ( ALT) etc.
1 ) Serum Bilirubin : It is the breakdown product of haemoglobin and a
major pigment in bile .
Normal Range
Indirect Bilirubin : 0.3-1.3 mg/dl
Direct Bilirubin : 0.1- 0.4 mg /dl
Significance :
high level indicates haemolysis , jaundice ( liver problem ) .
2) Serum (plasma ) protein : Albumins and globulins proteins are
major protein of plasma and produced by Liver .
Normal Range
Albumins : 3.5-5.1 gm/dl
globulins : 1.8-3.1 gm /dl

Significance
Increased level indicates dehydration Liver problems .
Deceased level Indicate Oedema, haemorrhage , Increased protein break
down .
3) alkaline Phosphatase ( ALP) : This enzyme produced in liver ,
bones , small intestine and kidneys , It catal yses splitting of phosphate group
from monophosphoric ester .
Normal range : 29-92 IU/L
High level indicates Rikets , Osteomlacia , abnormal absorption of vitamin D .

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Low level indicates Hypophosphatacia .
4) Serum Glutamic Oxaloacetic transaminase (SG OT) or
Aspartate transaminase (AST) : this enzyme is produced by liver
and it helps in energy production .
Normal range : 0.40 U/L

significance :
Increased level of SGOT indicates liver disease ( hepatitis , cirrhosis )
5) Serum Glutamate Pyruvate Transaminase ( ALT) : this
enzyme produced by liver and helps in formation of alanine .
Normal Range : 5-36 U/L
significance : Its increased level indicates Liver cell damage .

Serum Cholesterol test( Lipid Profile )

: this test is performed to check triglyceride and Cholesterol level in blood .
Normal Range
a) Total cholesterol level is < 200 mg /dl
b) LDL cholesterol level < 100 mg/dl
c) HDL Cholesterol level is equal or more than 60 mg/dl
d) Triglycerides level is < 150 mg/dl

Significance :
increased level of LDL and triglyceride indicates the risk of heart diseases ,
blockage of arteries .

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Chapter 13
Introduction to Pathology of Blood and Urine

Pathology of Blood
the study of blood related to diseases , causes and progression is called Pathology of blood .
The tests are performed for pathology of blood are called " Haematological Tests "
Different types of tests are performed for pathology of blood are following :
1) Haemoglobin Test : (Hb test )
This Test is performed to check Haemoglobine level of the body .

Normal Range
Male : 13.2 to 16.6 g/dl
Female : 11.6 to 15 g /dl
Children 6-12 years : 11.2 to 14. 5 g /dl

Significance and interpretation

Normal level is necessary for enough oxygen supply to tissues and transport
of CO2 to lungs .
Low level than normal range indicates Anaemia , Leukemia . and High level of
haemoglobin indicates polycythemia .( High concentration of RBCs).
2) RBCs Count : RBCs Contain Haemoglobin which transports O 2 and CO 2 , it
hlps in Blood clotting . This Test is performed to check Count of RBCs in blood

Normal range
Male : 4.5- 5.5 million/mm 3
Female : 3.5 - 5.5 million /mm 3
Children : 4.0-5.5 Million /mm 3
Significance :
Low level of RBCs than normal range in blood indicates Anaemia and
leukemia .
High rage of RBCs indicates Polycythemia .

3) Hematocrit (Hct) :
this test Indicates percentage of RBCs in whole blood sample. For example
40% Hct indicates that a 100ml sample has 40ml of red blood cells .

Normal range
Male : 40-50%
Female : 37 - 47 %

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Reason for high RBCs , High haemoglobine , and hematocrit .
1) Dehydration
2) lake of Oxygen ( due to lungs problems , smoking )
3) Kidney problems ( kidneys makes a hormone erythropoietin which trigger
the Bone marrow to make more red blood cells ) .

Reason for low RBCs , High haemoglobine , and hematocrit .

1) Heavy Blood loss .
2) Bone marrow failure .( Infection , tumor )
3) Any kind of Cancer treatment .
4) leukemia
5) Haematolysis ( break down of RBCs )
6) Kidney problems
4) White Blood Cell or Total Leucocyte Count (TLC) : This test is performed to
check conditions like : Infection , allergic reaction , Inflammation , Blood
cancer ( leukemia

Normal Range
male : 4500 to 11000 / ml
Female : 4500 to 11000 / ml

Significance :
 High Level of WBCs indicates Infection like( Appendicitis (
inflammation of appendix ) , Leukemia , Pneumonia , , meningitis etc. )
Allergic reaction , Inflammation , Blood cancer.
 High level may be due to theses reason also : Certain drugs ( antibiotics )
,Smoking , Removal of spleen , blood cancer , Inflammation .
 Low level of WBCs is called Leucopenia it indicates typhoid , Hepatitis ,
influenza , measles , anxiety .
 Low level of WBCs is may be due to these reasons also Bone marrow
problems , Cancer treatment , certain viral illness etc.

5) Different Leucocyte Count : (DLC )

Neutrophils 60-70%
Granulocytes Eosinophils 1-4%
Basophils 0-2%
Monocytes 5-10%
Lymphocytes 20-30%
Significance :
 High Neutrophils count(Neutrophilia) indicates severe bacterial
infection , whooping Cough ,UTI ( Urinary Tract Infection )

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 High Eosinophils Count ( Eosinophilia ) Indicates Intestinal Parasites ,
allergic condition .
 High basophils Count indicates granulocyte leukemia .
 High monocyte Count indicates bacterial Infection like malaria , and
Monocytic Leukemia .
 High Lymphocite Count indicates bacterial , viral or other infection(
whooping cough , mumps , Influenza , tuberculosis ) autoimmune
disorder .
6) Erythrocyte Sedimentation Rate ( ESR )
It indicates the rate at which eryrocytes sediment ( settle ) to the bottom ,
this rate of settlement is called RSR . the distance they cover in 1 hour is
recorded .
Normal range
Male : 0-15 mm/hr
female : 0-20 mm/hr

significance :
This test is performed to estimate inflammation in body , or infection .
if it is more than normal range indicates Inflammation in the body .

7) Platelets Count : they are necessary for blood clotting

Normal range
1.5 lac to 4.5 lac

Significant :
 this test is performed to check number of platelets . and before surgery
procedure this test is performed
 Low range than normal indicates bone marrow problems , leukemia , or
infection such as Hepatitis C , Dengue fever , Chikungunya , HIV . etc
a minor reduction occurs in pregnancy .
 High count of Platelets indicates Bleed ing , Cancer, iron Deficiency
,bone marrow problems .

8) Coagulation or Blood Clotting Time :

this test indicates the time blood takes to clot .
Normal Range
1) Slide method : 2-6 minutes
2) Capillary method : 2-6 minutes

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Significance :
Increased clotting time indicates Haemopholia , Vitamin K Deficiency ,
increased heparin level and pneumonia .
Decreased clotting indicates thrombosis and embolism .

9) Blood Sugar level

Normal range
Fasting : 70 - 99 mg /dl
after two hrs meals : less than 1 80 mg /dl
Random : 110-140 mg/dl

10) Blood Uric acid test :

Normal range
2-6 mg /100 ml
Significance : High uric acid indicates gout .renal failure .

Lymphocytes and platelets - Role in health in diseases

Lymphocytes
Lymphocytes are a type of WBC and a part of immune system . they are found
in lymph that is why they are called lymphocytes , They are 20 - 30 % of WBCs .
there are two main types of lymphocytes B lymphocytes ( B-Cells ) and t -
lymphocytes
( T -Cells )
B- lymphocytes produces antibodies that attack invading bacteria , viruses and
toxins
t- cells destroy infected body' s cells ( host cells) destroy tumor cells and
activate other immune sales
Count of lymphocytes
B- lymphocytes : 100-600 cells / µL of blood .
T- lymphocytes : 500-1200 cells / mm3 ( Cubic milliliter )
Functions
1) They Help immune system .
2) they produce antibodies .
3) they fight against foreign substances ( bacteria , viruses etc. )
4) they destroy infected cells .

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5) they kill tumor cells .
Abnormal Lymphocyte Count
an increase in lymphocyte count more than (4000 / µ L) in an adult , it is called
lymphocytiosis .
High Lymphocyte count indicates any of following condition :
1) infection
2) cancer of lymphocytes ( Lymphocytic leukemia )
3) autoimmune disorder .
Platelets
Platelets or thrombocytes are small and colourless blood cells . They are formed
by bone marrow , They mainly promote blood clotting and stop bleeding .
Role of platelets
1) Vasoconstriction : they secrete serotonin which causes constriction of blood
vessels .
2) Blood Coagulation : it plays an important role in blood clotting .
3) Fibrinolysis : Platelets help in breakdown of the clot and healing of damage
blood vessels .
Abnormal Platelets Count
Blood Platelet count below normal range is called Thrombocytopenia when high
level is called Thrombocythemia .
Low level leads to
 improper blood clotting
 Prolonged bleeding from cuts
 Usually heavy menstrual flows
 Blood in urine or stools
 Fatigue
 Bleeding from gums or nose
High level of platelets can cause dangerous clots in blood vessels , increasing
risk of stroke or heart attack .
Role of blood cells in health and Diseases

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Erythrocytes - Abnormal Cells and their Significance

Normal RBCs are round , disc-shaped , biconcave cells which don't have nucleus
.
Different type of abnormalities occurs in RBCs , some of them are :
1) Variation in size
2) Variation in Shape
3) Alteration in colour
Variation in size
a normal erythrocyte has an average diameter of 7.2 µ m ( 6.8 - 7.5 µ m ) . the normal size
cells are called normocyte .
anisocytosis a common term is used for an increased RBCs cells size .
Variation in Shape
Variation in Shape of a mature RBC have the shape other than normal Round ,
Disc shape and biconcave appearance . it is termed Poikilocytosis .
Poikilocytosis may be different types like Acanthocytes , Blister Cells etc.
Alteration in colour
the normal RBCs have a red color , which is provided by haemoglobin , the
alteration in colour is occurs with the change in concentration of Heamoglobin .
Note : any type of abnormality disturb the function of Erythrocytes .
Significance of Erythrocytes
1) transport of Oxygen from lungs to tissues .
2) transport of carbon dioxide from tissues to lungs .
3) transport of wastes from tissues to kidney .
4) Dilation of blood vessels : it release the ATP which cause relaxation of vessels
5) Immunity : when bacteria break the haemoglobin it release free radicals that
rupture the bacterial cell wall and membrane which causes death of bacteria .
Disorder of Erythrocytes and their significance
1) Anaemia : deficiency of RBCs or Haemaglobin in blood .
a) Iron deficiency anaemia

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b) magaloblastic anaemia ( Vit. B 12 and B 9 deficiency )
c) pernicious anaemia ( vit. B 12 deficiency )
d) Haemolytic anaemia
2) polycythemia ( increased RBCs count )
3) Hemoglobinopathies ( sickle cell means misshape of RBCs )
4) Red cell enzyme deficiency ( G6PD )
5) Hemochromatosis ( iron overload , it can poison organs and lead to cancer )

Normal Constituents of Urine

Urine is an aqueous solution , a by-product of the body, formed by the kidneys

and excreted out through urethra by a process called micturition .
Normal urine have 95% of water and 5% of other constituents .
Normal constituents of Urine
1) urea ( 9.3 gm / lt)
2) Chloride ( 1.87 gm /L )
3) Sodium ( 1.17 gm /L )
4) Potassium ( 0.750 gm /L )
5) Creatinine ( 0.670 gm / L )
6) Some dissolved ions and other compounds ( hormones , oxalic acid , hippuric
acid etc.)

Abnormal constituents of urine and their significance

SN Abnormal Constituents Significance diseases
1 Proteins Proteinuria
2 Sugar Glycosuria
3 Ketone bodies Ketonuria
4 Bile pigments and salts Jaundice
5 Blood Haematuria
6 Pus Pyuria

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