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Download MBBS Biochemistry PPT 50 Hemoglobin Metabolism Lecture Notes

Download MBBS (Bachelor of Medicine, Bachelor of Surgery) 1st year (First Year) Biochemistry ppt lectures Topic 50 Hemoglobin Metabolism Notes. - biochemistry notes pdf, biochemistry mbbs 1st year notes pdf, biochemistry mbbs notes pdf, biochemistry lecture notes, paramedical biochemistry notes, medical biochemistry pdf, biochemistry lecture notes 2022 ppt, biochemistry pdf.

This post was last modified on 05 April 2022

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vHeme Synthesis

qPorphyrias (Disorders Of Heme Synthesis)

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vGlobin Synthesis

qAbnormal Hb variants/
Hemoglobinopathies(Disorders of Globin Synthesis)

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Hemoglobin Breakdown

qFormation and Fate of Bilirubin
qHyperbilirubinemia

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Jaundice : Causes Types and Diagnosis

Hemoglobin Biosynthesis
Hemoglobin biosynthesis

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includes biosynthesis of:

vHeme
vGlobin Polypeptide chains

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Amount of Hb biosynthesized=

6.25 gm/day
Site For Hemoglobin

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Biosynthesis

Organs Involved In

Hb Biosynthesis

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Bone Marrow-
Immature Erythrocytes ? 85%
Liver ? 15 %
Requirements For

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Hemoglobin Biosynthesis

Normal biosynthesis of

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Hemoglobin depends upon an

Quality and Quantity of :

qAmino acids

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qMinerals
qVitamins
Heme Biosynthesis

OR

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Porphyrin Pathway

Biosynthesis

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Of

Ferroprotoporphyrin


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Site For Heme Biosynthesis

Organs

Bone Marrow-

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Immature Erythrocytes ? 85%
Liver ? 15 %

Cellular Site

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Mitochondrial Matrix
Cytosol
Requirements For

Heme Biosynthesis

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Metabolic Precursors for Heme Biosynthesis:

? Glycine and Succinyl-CoA

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Vitamins (5 Hematopoietic Vitamins):

vPantothenic acid (Vitamin B5)
vPyridoxine (Vitamin B6)
vFolate (Vitamin B10)

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vCyanocobalamin (Vitamin B12)
vVitamin- C (Ascorbic acid)

Minerals for Heme Biosynthesis:

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?Iron ( Fe++)
?Copper (Cu++)
?Zinc ( Zn ++)
Stages and Steps

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Of

Heme Biosynthesis

3 Stages Of Heme Biosynthesis

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1. Synthesis of -Amino Levulinic Acid

( ALA)

(In Mitochondrial Matrix)

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2. Synthesis of CoproPorphyrinogen?III

( CPG-III)

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(In Cytoplasm)

3. Synthesis of ProtoPorphyrin IX and

Incorporation of Fe++ to Form Heme

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(In Mitochondrial Matrix)

7 steps in Heme Biosynthesis

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Step 1 in Mitochondrial Matrix
Steps 2,3,4 in Cytoplasm
Steps 5,6 and 7 in

Mitochondrial matrix

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?Important Intermediates of Heme

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Synthesis Pathway:

-Aminolevulinic acid

(ALA)

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Porphobilinogen

(PBG = Pyrrole derivative)
Uroporphyrinogen III

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( UPG? Heme precursor)
Protoporphyrin IX

(Direct Heme precursor)

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-Aminolevulinic Acid (ALA)

? Synthesis of Heme starts in Mitochondrial matrix
? Succinyl-CoA and Glycine undergo a condensation ALA
? Reaction is catalyzed by enzyme ALA Synthase

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? ALA Synthase requires

?Vitamin B6 (PLP)
? Copper ions

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PLP used in first step of

Heme biosynthesis

activates Glycine.

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Presence of free Heme inhibits

the synthesis of an enzyme

- ALA Synthase.

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This represents a Feedback

mechanism for Heme synthesis.

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This first step is a Rate limiting

step of Heme synthesis:

vStimulated by the presence of

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Globin chains.

vInhibited by the presence of free

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Heme groups.


Rate of Heme

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biosynthesis has good

coordination with

Globin synthesis.

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Porphobilinogen (PBG)

? ALA leaves the Mitochondria Reach Cytoplasm
? 2x ALA condense together to form Porphobilinogen (PBG).

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? Reaction is catalyzed by Porphobilinogen Synthase
/(ALA dehydratase)
Mitochondrial -Aminolevulinic acid

(ALA) is transported to the cytoplasm,

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where

ALA Dehydratase /Porphobilinogen

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Synthase- Zinc containing enzyme.

Dimerizes 2 molecules of ALA to produce
The Pyrrole ring compound is

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Porphobilinogen (PBG).

PBG then biosynthesizes Porphyrin

ring.

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The reactions involved for its

synthesis are extremely complex

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but can be summarized as follows:
The condensation of four PBG

molecules

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Form an asymmetric cyclic

Uroporphyrinogen III(UPG III).

Synthesis of UPG III requires the

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presence of two enzymes:

v Uroporphyrinogen I Synthase
v Uroporphyrinogen III Cosynthase

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During UPG synthesis there

involves the formation of

short-lived intermediate

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Hydroxy Methyl Bilane

(HMB).

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UPG I Synthase/PBG

Deaminase /HMB Synthase

Transforms 4 molecules of PBG

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to linear Tetrapyrrole

Hydroxy Methyl Bilane

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(HMB)
HMB spontaneously

cyclizes to form UPG III

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by UPG III Cosynthase

Conversion of

Uroporphyrinogen II Coproporphyrinogen II

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4 Acetate residues of

Uroporphyrinogen III are

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Decarboxylated into 4 Methyl

groups Coproporphyrinogen III

Coproporphyrinogen III returns to

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the Mitochondria again.
UPG III is converted to Coproporphyrinogen III

(CPGIII) by Decarboxylation of the Acetate side

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chains

To Methyl groups under the influence of the

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enzyme Uroporphyrinogen Decarboxylase.
CPG III enters the Mitochondria where it is

converted to Protoporphyrinogen IX (PPG IX) by an

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unknown mechanism.

This reaction is catalyzed by the enzyme

Coproporphyrinogen Oxidase.

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Coproporphyrinogen III Protoporphyrinogen IX

In Mitochondria CPG III is oxidized to

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PPG IX.

Two Propionyl residues transformed

to Two Vinyl residues.

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Removal of two CO2 molecules.
Reaction catalyzed by CPG Oxidase.
Protoporphyrinogen is a

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colorless compound

It contains Methylene

bridges in tetrapyrrole

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ring structure.

Methylene bridges of

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Protoporphyrinogen

are oxidized to

Methenyl bridges to

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form ProtoPorphyrin
Protoporphyrinogen IX Protoporphyrin IX

? Oxidation of protoporphyrinogen IX produces the

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conjugated Methenyl bonds of Protoporphyrin IX

Final Formation of Heme

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? Fe2+ is incorporated into Protoporphyrin IX

? Reaction is catalyzed by enzyme Ferrochelatase/Heme

Synthase to Form Heme.

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Iron is chelated within Porphyrin

ring to form Heme by catalytic

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activity of Ferrochelatase.

Heme is incorporated into

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Proteins to become biologically

functional Hemoproteins.


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Heme

MetalloPorphyrin /Ferroprotoporphyrin

Heme forms various: Hemoproteins

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Hemoglobin
Myoglobin
Cytochromes
Catalase and Peroxidase

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Tryptophan Pyrrolase


Chlorophyll is a Magnesium

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containing Porphyrin present

in plants.

Chlorophyll is involved in

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photosynthesis of plants.
Regulation Of Heme Biosynthesis

ALA Synthase is an

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Allosteric regulatory

Enzyme of Heme

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biosynthetic pathway.
ALAS 1 occurs in Hepatocytes
ALAS 2 is found in Erythroid tissue

Rate of Heme biosynthesis is

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flexible.

Heme biosynthesis changes

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rapidly in response to a wide

variety of external stimuli.
Mechanisms and Factors

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Regulating Heme Biosynthesis

Feed Back Inhibition
Repression of ALA Synthase
Inhibition of transport of ALA

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Synthase from Cytosol to

Mitochondrial matrix.

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Erythropoietin levels
Iron levels
ALA Synthase is a key regulatory

enzyme of Heme biosynthesis.

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It is an allosteric enzyme that is

inhibited by an end product-Heme

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(Feedback inhibition)

Requires Pyridoxal Phosphate as

a coenzyme

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Erythropoietin Stimulates

Heme Biosynthesis
Erythropoietin is a Protein

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produced by Kidneys .

Erythropoietin stimulates

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the ALA Synthase activity.

Erythropoietin Synthesis

increased in high altitude

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dwellers.

Erythropoietin decreased

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in chronic renal failure.

(Associated with Anemia)
Iron Levels Affect Heme Synthesis

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The amount of cellular

Iron determines

The affinity for Iron

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Responsive Element-

Binding Protein(IRE-BP).
When Iron levels are low

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There is a high binding

affinity of IRE-BP with

mRNA of ALA.

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Which serves to inhibit the

translation of ALA mRNA

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Results in decrease of Heme

biosynthesis.

When Iron levels are

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sufficient.

There is a low binding affinity

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of IRE-BP with mRNA of ALA.

Thus allowing translation of

ALA mRNA

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Stimulation of Heme synthesis.


How Iron Levels Affect

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Heme Synthesis

Iron and Hemoglobin

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Iron deficient red

blood cells

Low number of red

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blood cells

Note the hollow and

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blanched appearance

of the red blood cells.
If either Heme or Globin

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synthesis is impaired

Iron is not utilized and

accumulates in the RBC.

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When Heme

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biosynthesis is impaired

Iron is underutilized
Mitochondria or Nucleus

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of Erythroblasts become

encrusted with Iron.


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A Sideroblast is

a Nucleated Erythrocyte

Containing Iron granules in

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its cytoplasm in the bone

marrow.

Sideroblast

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Sideroblast is

an Erythroblast with Iron granules

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(Pappenheimer bodies) seen in

bone marrow stained by
Prussian blue or Perl stains.
A precursor Red blood

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cell (Immature RBC)

with a ring of Iron

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around the nucleus is

called a ringed

Sideroblast.

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Siderocyte

Siderocyte is a non-

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nucleated red cell with

Iron granules
(Pappenheimer bodies)
A mature RBC with an

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accumulated Iron is

termed as Siderocyte .

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Siderocytes

are abnormally increased in :

Sideroblastic Anemia

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Hemosiderosis
Hemoglobinopathies


The Iron within

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Sideroblasts and

Siderocytes can be

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visualized by staining with

Prussian blue stain.


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Ringed Sideroblast


RINGED SIDEROBLASTS AND

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SIDEROCYTE

Effect Of Drugs and Other

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Substances On Heme Biosynthesis
Certain Drugs and

Steroid Hormones

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increases Heme

biosynthesis.

Ingestion of drugs like

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Phenobarbitals ,Insecticides,

certain chemical carcinogen,

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Markedly increases ALA

Synthase (ALAS1 of Hepatocytes)

activity.

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This increases production of Heme.
The biosynthesized Heme in

response to drug administration

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Is used for production of

Cytochrome P450

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Cyt P450 ?A Hemoprotein is

responsible for drug detoxification.

Lead Poisoning Affects

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Heme Biosynthesis

and

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Causes Anemia
Following Enzymes of Heme biosynthetic

pathway are inhibited by Lead ions

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(Pb2+) in cases of Lead poisoning.

ALA Dehydratase / Porphobilinogen

Synthase

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Ferrochelatase / Heme Synthase

Thus Lead

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poisoning

Inhibit Heme

biosynthesis and

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Leads to Anemia.
Porphyrins

Porphyrins are

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chemically cyclic tetra

Pyrrole ring structures

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with substituted

groups.
Porphyrins has

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conjugated ring system

Alternate single and

double bonds (Methenyl

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bonds).

Porphyrins are colored and

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Fluorescent compounds with

Methenyl bridges/ Methyne

bonds in it.

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The double bonds in Porphyrins

absorb visible light and appear

colored compounds.

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The Conjugated bonds of

Porphyrins in UV light shows

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fluorescence intense reddish

pink color.

Types Of Porphyrins

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Based on arrangements of

Substituted groups on

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Tetrapyrrole Rings
Types Of Porphyrins

Type I Porphyrins
Type III Porphyrins

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Type I Porphyrins

has symmetric

arrangements of

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substituted groups

in tetra pyrrole ring

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structure.

Type III Porphyrins

have asymmetrical

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distribution of the

substituted groups in

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tetra pyrrole rings.
Type III Porphyrins

are most predominant

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in biological system.

ProtoPorphyrin IX is of

type III Porphyrins

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Fischer placed

ProtoPorphyrin in 9th

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series of 15 possible isomers.
In Disorders of Porphyrias

Porphyrins are

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abnormally elevated

In blood and excreted in

urine.

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Porphyrias

Gk- Porphyria= Purple
Porphyrias

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Disorders Due To

Defective

Heme Biosynthesis

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Porphyrinurias
What Are Porphyrias?

The Porphyrias are

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group of disorders

Associated to

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defective Heme

biosynthesis.
Basic Cause Of Porphyrias

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Metabolic block in

Heme biosynthesis

leads to Porphyrias.

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Defect in any one

Enzyme of Heme

biosynthesis

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Defect in Enzyme of Heme

Biosynthesis may be:

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?Inherited

?Acquired
Most of the Porphyrias

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are of Autosomal

Dominant inheritance .

Types and Classification

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Of Porphyrias
6 Common Types Of Porphyrias

S.No Type Of Porphyrias

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Enzyme Defect

1

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Acute Intermittent

UPG I Synthase/

Porphyria (AIP)

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PBG Deaminase

2

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Erythropoietic

UPG III Cosynthase

Porphyria

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3

Cutanea Tarda

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UPG Decarboxylase

4

Coproporphyria

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CPG Oxidase

5

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Variegate Porphyria

PPG Oxidase

6

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Protoporphyria

Ferrochelatase
Pneumonic To Remember

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6 Type Of Porphyrias

All Elephants Can

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Catch Ved Pathak.

Mitochondria

PORPHYRIAS

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GLYCINE + SuccinylCoA

Agent Orange

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ALA Synthase

d Aminolevulinic acid(ALA)

3p21/Xp11.21

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ALA-Dehydratase

ALA Dehydratase

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Deficiency

9q34

Porphobilinogen (PBG)

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Porphyria
Acute intermittent

PBG Deaminase

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Porphyria

Hydroxymethylbilane

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11q23

Uroporphyrinogen III

Congenital Erythropoietic

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Cosynthase

Porphyria

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Uroporphyrinogen III

10q26

Uroporphyrinogen

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Porphyria

Decarboxylase

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Cutanea tarda

Coprophyrinogen III

1q34

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Coproporphyrinogen

Herediatary

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oxidase

coproporphyria

Protoporphyrinogen IX

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9

Protoporphyrinogen

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Variegate

Protoporphyrin IX

Oxidase

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Porphyria

1q14

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Ferrochelatase

Erythropoietic

Heme

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18q21.3

protoporphyria
Different Basis For

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Classification Of Various

Types Of Porphyrias
On Basis Of Cause

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Primary/Congenital Porphyrias
Secondary/Acquired Porphyrias

On Basis Of Organ

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Hepatic Porphyria
Erythropoietic Porphyria
Inherited Porphyria

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Erythropoietic Porphyria - results from

excessive production of Porphyrins in the

bone marrow.

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Hepatic Porphyria - results from

excessive production of Porphyrins in the

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Liver.

Acquired Porphyria

Lead Intoxication - interferes with

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Protoporphyrin synthesis

?Chronic Alcoholic Liver disease

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On Basis Of Symptoms

Neurological Porphyrias/Acute Porphyrias

Acute Intermittent Porphyria

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Coproporphyria

Variegate Porphyria

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?Autonomic Dysfunction
?Abdominal pain
?Chest pain
?Confused Thoughts
?Depression and Psychosis

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Photosensitive Porphyria/Chronic

Porphyria

q Erythropoietic Porphyria

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q Cutanea Tarda

Porphyrins below skin exposed to

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sunlight shows

vRedness
vSwelling
vItching

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vBurning Sensation

Biochemical Alterations

And

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Consequences Of Porphyrias
Enzyme Defect Of Heme

Pathway

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Blocks the Reaction Of Heme Pathway

Accumulates

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Porphyrins

Intermediates of

Heme Pathways

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High levels of Porphyrin in blood

,Tissues and Urine (Porphyrias)
Porphyrinogens are oxidized

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to Porphyrins.

Porphyrins are coloured

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pigments.

Porphyrins are more stable

products.

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Accumulate in blood, tissues

and get excreted out through

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urine

Effects of Accumulated

Porphyrins and their Precursors

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?Porphyria Sufferers Shows

qSevere Anemia
qNeurological Disturbances
qExtreme sensitivity to sunlight

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Porphyria sufferers has no

normal Heme biosynthesis.

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No normal Hb to transport

Oxygen to cells.

Hence suffer from Anemia.

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Accumulation of

Porphyrinogens

in Brain and Skin can lead to:

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qNeurological symptoms

q Photosensitivity

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Enzyme block early in

Porphyrin pathway prior to

formation of Porphyrinogens.

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Accumulates ALA and PBG

Exhibits abdominal pain and

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neuropsychiatric symptoms.
Enzyme block occur later in

the Porphyrin pathway

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Accumulates Porphyrinogens-

CPG/PPG beneath skin.

Causes Photosensitivity

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when Porphyrins exposed to

light about 400 nm.

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The Porphyrins have

no useful function

Act as highly reactive

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oxidants, damaging to

tissues.
Porphyrins get excited at

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400 nm.

Shows sharp absorption band

--- Content provided by FirstRanker.com ---

near 400nm (Soret band)

Porphyrins reacts with

molecular Oxygen

--- Content provided by⁠ FirstRanker.com ---


To form highly reactive

oxygen free radicals.

--- Content provided by​ FirstRanker.com ---

Injure Lysosomes and other

cellular organelles.
Damaged Lysosomes release

--- Content provided by​ FirstRanker.com ---

their degradative enzymes .

Causing variable degrees of

skin damage including

--- Content provided by FirstRanker.com ---


scarring.

Porphyrias are characterized by

--- Content provided by​ FirstRanker.com ---

?Extreme sensitivity to light

(exposure to sunlight causes

vesicular erythema),

--- Content provided by​ FirstRanker.com ---


? Excretes reddish-brown urine
? Possess reddish-brown teeth,

and ulcers

--- Content provided by⁠ FirstRanker.com ---

Destruction of cartilage and

bone

Causing the deformation of the

--- Content provided by‌ FirstRanker.com ---


nose, ears, and fingers.

Mental aberrations, such as

--- Content provided by⁠ FirstRanker.com ---

hysteria, manic-depressive

psychosis, and delirium.

Porphyrias are cruelly referred to

--- Content provided by‌ FirstRanker.com ---


as a Vampire's disease.

Thought to be a cause of the

--- Content provided by FirstRanker.com ---

madness of King George III.

Can be caused by lead poisoning:

The fall of the Roman Empire!

--- Content provided by‍ FirstRanker.com ---





Porphyria

--- Content provided by‍ FirstRanker.com ---

Diagnosis Of Porphyrias

Porphyrias are rare, but

frightening conditions:

--- Content provided by⁠ FirstRanker.com ---


Hard to diagnose and there is

no cure for Porphyrias.
Porphyrins Excreted

--- Content provided by FirstRanker.com ---


In Urine and Feces

Uroporphyrin excreted in

--- Content provided by​ FirstRanker.com ---

urine.

ProtoPorphyrin excreted in

feces.

--- Content provided by FirstRanker.com ---


Coproporphyrin excreted

either in urine /feces.

--- Content provided by⁠ FirstRanker.com ---

Porphyrins are Colored

and Fluorescent.
Porphyrias are diagnosed by analysis of

--- Content provided by FirstRanker.com ---

Porphyrins in the laboratory.

Spectrophotometry
Fluorometry

--- Content provided by​ FirstRanker.com ---

Woods lamp- Fluorescence in aqueous

layered viewed.

Based on quantitative Ehrlich's reagent

--- Content provided by FirstRanker.com ---


Watson Schwartz
Hoesch Test

Defective enzymes of

--- Content provided by‍ FirstRanker.com ---


Porphyrias can be assayed

by various methods.

--- Content provided by​ FirstRanker.com ---

Enzyme Assay- HPLC.
Acute Intermittent

Porphyria

--- Content provided by​ FirstRanker.com ---

(AIP)

Acute Intermittent Porphyria

vThe most common type of

--- Content provided by⁠ FirstRanker.com ---


Porphyria.

vAutosomal Dominant trait.
vSymptoms more common in

--- Content provided by​ FirstRanker.com ---


females than males.
Acute Intermittent Porphyria

Type Of Porphyria-

--- Content provided by⁠ FirstRanker.com ---


Acute/Hepatic/
Neurological Porphyria

Enzyme Defect Of AIP

--- Content provided by‍ FirstRanker.com ---


UPG I Synthase/PBG Deaminase

Biochemical Alteration In AIP

--- Content provided by FirstRanker.com ---

No conversion of PBG to

UPG III.

PBG and -ALA are

--- Content provided by FirstRanker.com ---


abnormally elevated in

blood, tissues and urine.

--- Content provided by‌ FirstRanker.com ---

Manifestations Of AIP
ALA and PBG

accumulates in CNS.

--- Content provided by⁠ FirstRanker.com ---

This causes excitation of

visceral pain fibers

Leads to acute pain crises.

--- Content provided by FirstRanker.com ---


ALA blocks the

action of GABA.

--- Content provided by‍ FirstRanker.com ---

Possess neurological

symptoms.
Symptoms of AIP are Acute

--- Content provided by FirstRanker.com ---

and Intermittent.

Symptoms does not occur

before puberty and shown at

--- Content provided by​ FirstRanker.com ---


Adolescence.

(Due to Steroidal Hormone action)

--- Content provided by‌ FirstRanker.com ---

Person with AIP has

Affected GIT, Heart and Brain.

Abdominal colic pain

--- Content provided by‍ FirstRanker.com ---


No abdominal tenderness

Vomiting, Constipation

--- Content provided by‌ FirstRanker.com ---

Tachycardia, Hypertension

Neuro toxicity

Behavioral changes, seizures

--- Content provided by​ FirstRanker.com ---

AIP symptoms gets aggravated

during:

vInfections

--- Content provided by‌ FirstRanker.com ---

vFasting
vIntake of drugs

Diagnostic Test For AIP

--- Content provided by​ FirstRanker.com ---

Watson and Schwartz Test

using Woods lamp ( UV lamp)

Detects urine Porphobilin.

--- Content provided by‌ FirstRanker.com ---

Treatment of AIP

Infusion of Hematin
Represses ALA Synthase

--- Content provided by‌ FirstRanker.com ---

synthesis.

Administration Of

Glucose.

--- Content provided by​ FirstRanker.com ---


High cellular Glucose

prevents induction of ALA

--- Content provided by​ FirstRanker.com ---

Synthase.


Use of Sunscreens that

--- Content provided by FirstRanker.com ---

filter out visible light,

Can be used in

management of

--- Content provided by‍ FirstRanker.com ---


Photosensitive Porphyrias

The Madness of Inbreeding

--- Content provided by‌ FirstRanker.com ---

King George III : Severe abdominal pain, mental confusion,

dark urine.
Enzyme Defects Responsible for The Porphyrias

--- Content provided by​ FirstRanker.com ---

Type

Enzyme Involved

Major Symptoms

--- Content provided by‌ FirstRanker.com ---


Laboratory tests

Acute intermittent Uroporphyrinogen

--- Content provided by‌ FirstRanker.com ---

Abdominal pain

urinary Porphobilinogen

Porphyria

--- Content provided by​ FirstRanker.com ---


synthase

Neuropsychiatric

--- Content provided by FirstRanker.com ---

Congenital

Uroporphyrinogen

Photosensitivity

--- Content provided by FirstRanker.com ---


urinary uroporphyrin

Erythropoietic

--- Content provided by FirstRanker.com ---

III Cosynthase

Porphyria

Porphobilinogen

--- Content provided by​ FirstRanker.com ---


Porphyria cutanea

UPG

--- Content provided by‌ FirstRanker.com ---

Photosensitivity

urinary uroporphyrin

tarda

--- Content provided by FirstRanker.com ---


Decarboxylase

Porphobilinogen

--- Content provided by‍ FirstRanker.com ---

Variegate Porphyria

PPG Oxidase

Photosensitivity

--- Content provided by‍ FirstRanker.com ---


urinary uroporphyrin

Abdominal pain

--- Content provided by FirstRanker.com ---

Neuropsychiatric

fecal coproporphyrin
fecal Protoporphyrin

--- Content provided by‍ FirstRanker.com ---

Erythropoietic

Ferrochelatase

Photosensitivity

--- Content provided by​ FirstRanker.com ---


fecal Protoporphyrin

protoporphyria

--- Content provided by‌ FirstRanker.com ---

red cell Protoporphyrin
Globin Biosynthesis

The biosynthesis of Hemoglobin

--- Content provided by‌ FirstRanker.com ---

Globin Polypeptide chains is

under genetic control.

Using Protein synthetic machinery.

--- Content provided by‌ FirstRanker.com ---


Globin chain biosynthesis occurs

in cytosol on Polyribosomes.
Number And Types Of Globin

--- Content provided by‍ FirstRanker.com ---


Chains Biosynthesized

In Human Life

--- Content provided by FirstRanker.com ---

6 different types of Globin chains are

associated with Normal Hb variants :

Globin

--- Content provided by​ FirstRanker.com ---


Globin

Globin

--- Content provided by‌ FirstRanker.com ---

Globin

Globin

Globin

--- Content provided by FirstRanker.com ---

To biosynthesize these 6 types of

Globin chains

Human being normally carry

--- Content provided by FirstRanker.com ---


8 functional Globin genes


Arranged in two duplicate gene

--- Content provided by‌ FirstRanker.com ---


clusters.

Globin Gene Clusters
The -like cluster located on the

--- Content provided by​ FirstRanker.com ---


short arm of chromosome 11

The -like cluster is located on

--- Content provided by⁠ FirstRanker.com ---

the short arm of chromosome 16

Globin polypeptide

chain biosynthesis

--- Content provided by​ FirstRanker.com ---


begins in the yolk sac

At about 3 weeks' of

--- Content provided by‌ FirstRanker.com ---

gestation.
Ontogeny of Globin Synthesis

Ontogeny of Globin Synthesis

--- Content provided by‌ FirstRanker.com ---

Time

Region

Type of Globin

--- Content provided by​ FirstRanker.com ---


Normal Hb Variant

Gene

--- Content provided by‌ FirstRanker.com ---

Type of Hb

3 weeks of

Yolk Sac

--- Content provided by​ FirstRanker.com ---


&

(Hb Gower I ( )2

--- Content provided by FirstRanker.com ---

Gestation

5 weeks of

Yolk Sac

--- Content provided by FirstRanker.com ---


&

Hb Portland ( )2

--- Content provided by⁠ FirstRanker.com ---

Gestation

Hb G

ower II ( )

--- Content provided by⁠ FirstRanker.com ---


2

6-30 weeks of

--- Content provided by‌ FirstRanker.com ---

Liver & spleen

& &

Hb F ( )2

--- Content provided by FirstRanker.com ---


Gestation

30 weeks of

--- Content provided by‌ FirstRanker.com ---

Liver

Hb A2 ( )2

Gestation

--- Content provided by FirstRanker.com ---


At Birth

Bone Marrow

--- Content provided by⁠ FirstRanker.com ---

& &

Hb A ( )2

Hb F ( )2

--- Content provided by​ FirstRanker.com ---

Synthesis of Globin

Primary Structure Of

Globin

--- Content provided by FirstRanker.com ---


The primary structure of globin

refers to the amino acid

--- Content provided by⁠ FirstRanker.com ---

sequence of the various chain

types.

Numbering from the N-terminal

--- Content provided by‍ FirstRanker.com ---


end identifies the position of

individual amino acids.
The specific number

--- Content provided by‍ FirstRanker.com ---


,and sequence of amino

acids in Globin chains

--- Content provided by​ FirstRanker.com ---

Is very important for the

normal structure and

function of Hemoglobin.

--- Content provided by‌ FirstRanker.com ---


Secondary Structure of Globin

The secondary structure of all Globin

--- Content provided by​ FirstRanker.com ---

chain types comprised of:

9 Non-helical sections joined by 8

Helical sections.

--- Content provided by​ FirstRanker.com ---


The Helical sections of Globin Chains

are identified by the letters A-H
While the non helical are

--- Content provided by⁠ FirstRanker.com ---


identified by a pair of letters

corresponding to the adjacent

--- Content provided by⁠ FirstRanker.com ---

helices

e.g. NA (N-terminal end to the

start of A helix), AB (joins the A

--- Content provided by‌ FirstRanker.com ---


helix to the B helix) etc.

Tertiary Structure of Globin

--- Content provided by​ FirstRanker.com ---

The secondary structure is

further folded and bended on

its own to form 3 dimensional

--- Content provided by​ FirstRanker.com ---


subunit.

To form a Tertiary structure of

--- Content provided by⁠ FirstRanker.com ---

Globin.
Heme gets incorporated in

the Heme pocket formed

--- Content provided by⁠ FirstRanker.com ---

inside each Globin subunit.

Quaternary Structure Of Hb

It is native conformational

--- Content provided by​ FirstRanker.com ---


state of Hb/ Functional

form of Hb.

--- Content provided by FirstRanker.com ---


Four subunits of tertiary

structure ,non covalently

--- Content provided by‌ FirstRanker.com ---

linked

To form quaternary level

of organization of Hb.

--- Content provided by FirstRanker.com ---


Assembly Of Hemoglobin
Although Heme and Globin

synthesis occur separately within

--- Content provided by⁠ FirstRanker.com ---


developing red cell precursors,


Their rates of synthesis are

--- Content provided by⁠ FirstRanker.com ---


carefully coordinated to ensure

optimal efficiency of Hb

--- Content provided by‍ FirstRanker.com ---

assembly.

Synthesis of Hemoglobin


--- Content provided by FirstRanker.com ---

HEMOGLOBIN SYNTHESIS

Normal structure of Hb

includes the structure and

--- Content provided by FirstRanker.com ---


proportion of Globin chains


Which are necessary for the

--- Content provided by FirstRanker.com ---


normal function of

Hemoglobin
Decreased Concentration of

--- Content provided by⁠ FirstRanker.com ---


Hemoglobin in the red blood

cells

--- Content provided by⁠ FirstRanker.com ---

Caused due to any abnormality
Results into a clinical

situation called Anemia

--- Content provided by FirstRanker.com ---

Mechanisms Regulating

Hemoglobin Synthesis
Formation of Hemoglobin is

--- Content provided by FirstRanker.com ---

regulated by several

mechanisms:

The rate of Globin

--- Content provided by‌ FirstRanker.com ---


biosynthesis is

directly related to the

--- Content provided by​ FirstRanker.com ---

rate of Heme

biosynthesis.
Heme stimulates

--- Content provided by FirstRanker.com ---

Globin biosynthesis by


Inactivating an

--- Content provided by FirstRanker.com ---

inhibitor of Globin

translation.

Negative feedback of

--- Content provided by FirstRanker.com ---


Heme.

High concentrations of

--- Content provided by FirstRanker.com ---

Heme

Prevent the mitochondrial

import of the first enzyme in

--- Content provided by‌ FirstRanker.com ---


Heme synthesis, ALA

Synthase ( ALAS).
The Concentration Of Iron

--- Content provided by​ FirstRanker.com ---


An Iron Responsive Element-

binding protein (IRE-BP)

--- Content provided by⁠ FirstRanker.com ---

binds to mRNA

Iron Response Elements (IRE-

BP) affects the translation of the

--- Content provided by‍ FirstRanker.com ---


mRNA for ALAS, Ferritin and

Transferrin receptors.

--- Content provided by‌ FirstRanker.com ---

Low Iron Levels =
Low Heme Synthesis

Sufficient Iron Levels=

--- Content provided by‌ FirstRanker.com ---

Adequate Heme Synthesis
Disorders Associated

To

--- Content provided by‌ FirstRanker.com ---

Globin Chain Synthesis

Of Hemoglobin

Hemoglobinopathies

--- Content provided by‌ FirstRanker.com ---


Caused By

Abnormal Hb Variants
What are Abnormal Hb variants ?

--- Content provided by‍ FirstRanker.com ---


Abnormal Hb variants are

Hemoglobin's with:

--- Content provided by​ FirstRanker.com ---

Normal Heme and Altered Globin Chain

Abnormal Hb variants are

structurally abnormal

--- Content provided by‌ FirstRanker.com ---


The Abnormal Hb variants may

be abnormal functionally
Approximately 400

--- Content provided by⁠ FirstRanker.com ---


abnormal Hb variants

are detected out.

--- Content provided by⁠ FirstRanker.com ---

But not all abnormal Hb

variants affect the

normal function of Hb.

--- Content provided by​ FirstRanker.com ---


Basic Cause For Formation Of

Abnormal Hb Variants
Mutations In Globin Genes

--- Content provided by‌ FirstRanker.com ---


Altered /Mutated

Globin Genes leads to

--- Content provided by‍ FirstRanker.com ---

form Abnormal Hb

variants.

Abnormal Hb Variants

--- Content provided by​ FirstRanker.com ---


Occurs due to Mutation

in Globin Genes.

--- Content provided by‍ FirstRanker.com ---

Leads to defective

Globin chain synthesis.
Formation Of

--- Content provided by FirstRanker.com ---

Abnormal Hb Variants

Leads To

Hemoglobinopathies

--- Content provided by⁠ FirstRanker.com ---


What are Hemoglobinopathies?
Hemoglobinopathies are

genetic disorders

--- Content provided by⁠ FirstRanker.com ---


associated to


Structurally and

--- Content provided by⁠ FirstRanker.com ---


Functionally Abnormal

Hemoglobin variants.

--- Content provided by‌ FirstRanker.com ---



Structurally and

Functionally Abnormal

--- Content provided by‌ FirstRanker.com ---


Hb variants in human

body leads to

--- Content provided by‍ FirstRanker.com ---

Hemoglobinopathies
Types Of

Abnormal Hemoglobin

--- Content provided by‌ FirstRanker.com ---

Variants

and

Hemoglobinopathies

--- Content provided by FirstRanker.com ---


Broadly two types

of Hb abnormalities
Qualitative

--- Content provided by FirstRanker.com ---


Hb Abnormalities:

Mutations in Structural

--- Content provided by‍ FirstRanker.com ---

Globin genes

e.g. HbS-Sickle cell anemia.

Quantitative

--- Content provided by FirstRanker.com ---


Hb Abnormalities:

Mutations in Regulatory

--- Content provided by‍ FirstRanker.com ---

Globin Genes

e.g. a Thalassemia

b Thalassemia

--- Content provided by​ FirstRanker.com ---

Qualitative

Abnormal Hb variants:

Caused due to mutations in structural

--- Content provided by‌ FirstRanker.com ---


Globin gene.

Has altered amino acid sequence in

--- Content provided by‍ FirstRanker.com ---

Globin polypeptide chain.

Has altered Globin subunits in Hb

structure

--- Content provided by‍ FirstRanker.com ---


But Has Normal Heme Structure.

Examples of Common

--- Content provided by​ FirstRanker.com ---

Abnormal Hb variants

And Corresponding

Hemoglobinopathy

--- Content provided by‌ FirstRanker.com ---


Due to Structural Globin Gene

Mutations

--- Content provided by⁠ FirstRanker.com ---

OR

Symptomatic Abnormal

Hb Variants

--- Content provided by FirstRanker.com ---

Abnormal Hb

Globin

Amino acid Altered

--- Content provided by​ FirstRanker.com ---


Variants

Gene/Chain

--- Content provided by‍ FirstRanker.com ---

In Globin Chain

Altered

Hb S

--- Content provided by FirstRanker.com ---


6 GLU VAL

Sickle cell Hb

--- Content provided by​ FirstRanker.com ---

Hb C

6 GLU LYS

Cooley's Hb

--- Content provided by‍ FirstRanker.com ---


Hb D

121 GLU GLN

--- Content provided by‌ FirstRanker.com ---

Punjab Hb

Hb E

26 GLU LYS

--- Content provided by‍ FirstRanker.com ---


Hb M

87 HIS TYR

--- Content provided by‍ FirstRanker.com ---

Hb has Fe+3

Proximal

If noted most common abnormal Hb

--- Content provided by⁠ FirstRanker.com ---


variants has:

Altered Globin genes and Globin

--- Content provided by⁠ FirstRanker.com ---

chains.

Substitution of Polar amino acid

"GLUTAMATE" with another amino

--- Content provided by​ FirstRanker.com ---


acid.
Non Symptomatic

Abnormal Hb Variants

--- Content provided by⁠ FirstRanker.com ---


Abnormal Hb Variants

?Hb P
?Hb Q

--- Content provided by⁠ FirstRanker.com ---

?Hb N
?Hb J
Consequences Of

Abnormal Hb Variants

--- Content provided by FirstRanker.com ---


Presence of

Symptomatic Abnormal Hb Variants

--- Content provided by⁠ FirstRanker.com ---

In RBCs

Alters normal structure and

function of Hb

--- Content provided by​ FirstRanker.com ---


Alters morphology of RBC's
Make RBC's fragile.
Causes Hemolysis, reduces Hb

--- Content provided by‍ FirstRanker.com ---

content and affects its function.

Leads to Hemolytic Anemia
Increases Unconjugated serum

--- Content provided by​ FirstRanker.com ---

Bilirubin

Causes Hemolytic Jaundice
Possess Splenomegaly -

--- Content provided by⁠ FirstRanker.com ---

Increased function of Spleen to

remove defective Erythrocytes

from the blood circulation.

--- Content provided by‍ FirstRanker.com ---


Detection Of

Abnormal Hb Variants
CBC and Blood Film Evaluation

--- Content provided by​ FirstRanker.com ---

Solubility Test
Electrophoresis
(Cellulose Acetate and Citrate Agar)
DNA Technology- PCR based

--- Content provided by⁠ FirstRanker.com ---

techniques:

DNA Finger Print Technique
Hybridization Technique

--- Content provided by‍ FirstRanker.com ---

Hemoglobinopathy-Antenatal

Diagnosis

Check the Test partners of

--- Content provided by⁠ FirstRanker.com ---


heterozygous or affected

individuals

--- Content provided by⁠ FirstRanker.com ---

Antenatal diagnosis from DNA

is obtained by chorionic villus

sampling, or by Amniocentesis

--- Content provided by​ FirstRanker.com ---

Common

Abnormal Hb Variant

Causing

--- Content provided by‍ FirstRanker.com ---


Hemoglobinopathy

Sickle Cel Hemoglobin (HbS)

--- Content provided by​ FirstRanker.com ---



Hb S is most

commonly occurring

--- Content provided by⁠ FirstRanker.com ---


abnormal Hb variant.
Hb S leads to Hemoglobinopathy

Sickle Cell Anemia

--- Content provided by‌ FirstRanker.com ---


Biochemical Defect TO Form HbS
Formation of

Sickle Cell Hemoglobin (HbS)

--- Content provided by‍ FirstRanker.com ---




Is a classic example of

--- Content provided by FirstRanker.com ---

point mutation

(Transversion)

Point mutation is a

--- Content provided by⁠ FirstRanker.com ---


substitution of

Nitrogen base in a

--- Content provided by‍ FirstRanker.com ---

normal Globin gene

sequence.
Substitution Of Nitrogen

--- Content provided by FirstRanker.com ---

Base Which Forms Hb S

Altered Nitrogen base

sequence in Beta Globin gene.

--- Content provided by​ FirstRanker.com ---


There is substitution of

Nitrogen base Thymine to

--- Content provided by⁠ FirstRanker.com ---

Adenine (T to A).

On transcription of mRNA it

has altered codon, GAG to

--- Content provided by⁠ FirstRanker.com ---


GUG

Altered amino acid

--- Content provided by​ FirstRanker.com ---

substitution in the beta Globin

chain.

Glutamate substituted by

--- Content provided by⁠ FirstRanker.com ---


Valine
On translation at 6th position

of Globin chain polar amino

--- Content provided by⁠ FirstRanker.com ---


acid GLU is substituted by

non polar amino acid VAL .

--- Content provided by‍ FirstRanker.com ---

This transforms HbA1 to HbS.

Deoxystate of HbS

Affects Solubility

--- Content provided by⁠ FirstRanker.com ---

HbS ? Is sickle cell Hb.
Altered HbS affects

the solubility of Hb

--- Content provided by FirstRanker.com ---

at Deoxystate in RBCs.

HbS at low oxygen tension /

deoxy state forms Deoxy HbS.

--- Content provided by​ FirstRanker.com ---


DeoxyHb S looses its polarity

/solubility

--- Content provided by‍ FirstRanker.com ---

Deoxy HbS forms protrusion on

the globin chain.

Sticky patch appears on HbS at

--- Content provided by FirstRanker.com ---


deoxy state.


Each Hb S fits into this

--- Content provided by‌ FirstRanker.com ---


complementary site of another

globin chain.

--- Content provided by​ FirstRanker.com ---

Many Hb S polymerizes inside

the RBC's

Forming a network of fibrous

--- Content provided by‌ FirstRanker.com ---


polymers.


HbS aggregates into long, rigid

--- Content provided by‌ FirstRanker.com ---


polymers are called Tactoids.

This makes HbS relatively

--- Content provided by‍ FirstRanker.com ---

insoluble and non functional.

EM of Red

Blood Cell

--- Content provided by‌ FirstRanker.com ---


showing

`Tactoids'
Tactoids stiffen and distort

--- Content provided by⁠ FirstRanker.com ---


the red blood cells.

RBC's changes morphology

--- Content provided by⁠ FirstRanker.com ---

and appear sickled/crescent

shaped.

Thus HbS Leads To

--- Content provided by FirstRanker.com ---


Sickling of Erythrocytes and

hemolysis.
Sickled Erythrocytes

--- Content provided by FirstRanker.com ---


may return to their

original shape when

--- Content provided by‌ FirstRanker.com ---

oxygenated.

Effect of Sickled RBC's

And its

--- Content provided by‍ FirstRanker.com ---


Associated Complications
HbS Causes

Reduction of RBC life span to

--- Content provided by FirstRanker.com ---


just 20 days.

Sickling of RBC's

--- Content provided by‍ FirstRanker.com ---

Distortion and lysis of RBC's

Hemolysis

Sickling distorts and make

--- Content provided by⁠ FirstRanker.com ---


RBC's fragile.

After several sickling

--- Content provided by‌ FirstRanker.com ---

episodes of RBC's

There is irreversible damage

to RBC membrane.

--- Content provided by‌ FirstRanker.com ---

Thus Sickling Of RBC's

causes Sickle Cell

Anemia/Hemolytic

--- Content provided by‌ FirstRanker.com ---


Anemia.

Sickled cells are phagocytized

--- Content provided by‍ FirstRanker.com ---

by macrophages.

In the spleen, Liver or bone

marrow.

--- Content provided by‌ FirstRanker.com ---

Sickling of RBC's

makes blood viscous

Lowers the rate of

--- Content provided by‌ FirstRanker.com ---


blood circulation.

Sickled cells has

--- Content provided by⁠ FirstRanker.com ---

increased tendency to

adhere to blood vessels.


--- Content provided by⁠ FirstRanker.com ---

Rigid sickled cells unable to

squeeze out through small

capillaries

--- Content provided by FirstRanker.com ---


Sickled cells get trapped in

small capillaries and block

--- Content provided by‌ FirstRanker.com ---

them.


Sickling produces localized

--- Content provided by FirstRanker.com ---

Anoxia/Tissue Hypoxia

Oxygen deprivation in the

tissues.

--- Content provided by​ FirstRanker.com ---


Lowers ATP production in

cells.

--- Content provided by‌ FirstRanker.com ---

Anoxia in turn leads to

increased sickling process.
Sickling causes pain

--- Content provided by FirstRanker.com ---

and infarction (death)

of cells in tissues.

Sickling causes

--- Content provided by‌ FirstRanker.com ---


Spleen Dysfunction

Making the spleen

--- Content provided by FirstRanker.com ---

non functional
Sickling Increases

susceptibility towards tissue

--- Content provided by‍ FirstRanker.com ---

infection

Premature death of

individuals before 20 years

--- Content provided by​ FirstRanker.com ---


due to infections.

Factors Increasing

--- Content provided by⁠ FirstRanker.com ---

Sickling Of RBC's
Extent of RBC's sickling

is related to

--- Content provided by⁠ FirstRanker.com ---

Amount of Hb S

present in Erythrocytes.

Conditions Creating Hb S in Deoxy

--- Content provided by​ FirstRanker.com ---


state:

Decreased pO2

--- Content provided by FirstRanker.com ---

Increased pCO2

Decreased pH

Increased 2,3 BPG

--- Content provided by⁠ FirstRanker.com ---


Dehydration


Sickle Cel Anemia

--- Content provided by​ FirstRanker.com ---


Sickle Cell Anemia is a genetic disorder

due to presence of abnormal Hb variant

--- Content provided by FirstRanker.com ---

HbS

It is a Commonest type of a

Hemoglobinopathy

--- Content provided by FirstRanker.com ---


It is a type of Hemolytic Anemia due to

Sickling of Erythrocytes
Inheritance of Sickle Cell Anemia

--- Content provided by FirstRanker.com ---


Sickle Cell Disease is an

Autosomal recessive

--- Content provided by‍ FirstRanker.com ---

disorder

Prevalence and Incidence

Prevalence of HbS

--- Content provided by FirstRanker.com ---


Tropical areas
Africa
South America

--- Content provided by FirstRanker.com ---

Incidence of HbS

1:5000 births.
Biochemical Defect

--- Content provided by‍ FirstRanker.com ---

To Cause

Sickle Cell Anaemia

Sickle Cell Anemia is caused by

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a point mutation in structural

beta Globin gene

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Characterized by the presence

of abnormal HbS in

Erythrocytes.

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Hb S in Deoxy state promotes

formation of hard, sticky,

sickled-shaped red blood

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cells ? Sickling of RBCs.

Types of Sickle Cell Anemia

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HbSS

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HbSS is sickle cell disease
Homozygous state
Full blown disease
100% HbS concentration.
Both Globin genes of 2

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chromosomes are

mutated.

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Globin chain has

alteration at 6 Glu to Val

HbAS

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HbAS is sickle cell trait
Heterozygous state.
50% HbA1 and 50 % HbS
Symptoms are mild and less

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severe.

Fatality can be delayed.
Sickle cell trait offers

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protection from

Malarial parasites-

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Plasmodium falciparum.

Hb SC Disease

Another red cell sickling disease

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Individual has mutant genes for

both Hb S and Hb C.

Has significant clinical variability

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Less severe anemia
Less painful crises.


Effects And Complications

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Of

Sickle Cell Anemia
Sickle Cell Anemia

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Leads To

Hemolytic Anemia
Hemolytic Jaundice

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Sickle Cell Anemia

Main Clinical Features

--- Content provided by FirstRanker.com ---

?Hemolysis
?Occlusion of blood

vessels by sickled red

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cells



Hemolysis /Lysis of Sickled RBC

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Low Oxygen transport to tissues

(Hemolytic Anemia)

Hemolytic Jaundice

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Tissue Hypoxia
Tissue Infarction
Tissue Infection
Painful Crisis
Fatality in severe cases

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Site of

Clinical Features Management

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Sickling
Bone

Painful crises

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Pain relief and hydration

Hydroxyurea

Lung

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Acute chest

Transfusion regimen, pain

--- Content provided by​ FirstRanker.com ---

syndrome

relief and hydration

Brain

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Stroke

Transfusion regimen.

--- Content provided by​ FirstRanker.com ---

Heart

Myocardial

Transfusion regimen, pain

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infarction

relief and hydration

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Spleen

Acute splenic

Transfusion, pain relief

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sequestration

and hydration

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Spleen

Hyposplenism

Pneumovax

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Retina

Proliferative

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Retinal surveil ance.

retinopathy

Laser

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`Tactoids' form at low oxygen

tension

Stiff Sickled red cells occlude

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small blood vessel
Tissue Hypoxia and Infarction
Tissue Infections
Symptoms are more severe
Fatality confirmed

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Sudden death during

intensive training

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Hematuria, Isosthenuria
(Renal Papillary Necrosis)
Infected RBC has incomplete life

cycle of parasites.

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Sickled erythrocytes efficiently

phagocytized and destroyed.

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Low K+ ion concentration in Hb S

is unfavorable for malarial

parasites to develop.

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Diagnosis of Sickle Cell Anemia

Sickling Test-

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Using Sodium Dithionite

reducing agent on blood

smear

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Watch microscopically for

sickled RBC's.
Sickle Cel Anemia ? Blood film

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Sickle

Cel s

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Erythroblasts

Howel -

Jol y Body

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Electrophoresis

Hb S is confirmed with

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Cellulose Acetate

Electrophoresis


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At Alkaline pH during

Electrophoresis.

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Hb S is less negatively

charged than Hb A1.

Hb S moves in a

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position between Hb

A1 and Hb A2.
Hb S migrates more slowly

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towards anode than Hb A1.

Altered mobility of HbS is due

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to absence of negatively

charged Glutamate residue.

Sickle Cell Anemia - Treatment

--- Content provided by FirstRanker.com ---


Adequate hydration
Analgesics to relive pain
Aggressive Antibiotic therapy

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to arrest the infection.
Ingestion of 0.01 M of

Potassium or Sodium

--- Content provided by​ FirstRanker.com ---

Cyanate.

Prevents sickling of RBC's and

its complications.

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Opiates and hydration

for painful crises

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Pneumococcal

vaccination

Retinal surveillance

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Hydroxyurea an antitumor drug
Used in therapy of Sickle cell

anemia.

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Increases circulating levels of Hb F
Decreases Sickling
Decreases painful crises
Reduces mortality

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Blood Transfusion

for serious

manifestations

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Support with Folate,

Iron chelation.

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Stem cell transplant
Thalassemias

Thalassemia's are

--- Content provided by FirstRanker.com ---

Hemoglobinopathies

Caused due to

defect/mutations in

--- Content provided by FirstRanker.com ---


Regulatory Globin

genes of Globin chain

--- Content provided by⁠ FirstRanker.com ---

synthesis.
Individual suffering from

Thalassemia's has

--- Content provided by​ FirstRanker.com ---

Structurally and

functionally unfavorable

abnormal Hb variants.

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Thalassemias are

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Autosomal recessive

blood disorders.
Thalassemias are

--- Content provided by‌ FirstRanker.com ---

Characterized with Anemia

Thalassemias mostly

occur in regions of

--- Content provided by FirstRanker.com ---


Mediterranean sea.

Also termed as

--- Content provided by‍ FirstRanker.com ---

Mediterranean Anemia.
Thalassemias are also

prevalent

--- Content provided by​ FirstRanker.com ---

In Arab,Americans, and

Asians

In populations where

--- Content provided by‌ FirstRanker.com ---


Malaria is endemic

Causes Of Thalassemias
Thalassemias due to

--- Content provided by‌ FirstRanker.com ---


Regulatory Gene mutations

is a quantitative abnormality

--- Content provided by‌ FirstRanker.com ---

of Hemoglobin.

Mutations in Regulatory

Genes of Globin chain

--- Content provided by‌ FirstRanker.com ---


synthesis.

Suppression of Globin

--- Content provided by​ FirstRanker.com ---

chain synthesis.
Reduced/Absent of one or

more of Globin polypeptide

--- Content provided by‌ FirstRanker.com ---

chain synthesis of Hb.

Globin chains has normal amino

acid sequence.

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Alpha Thalassemia ?

reduced alpha chain synthesis

--- Content provided by⁠ FirstRanker.com ---

Beta Thalassemia ?
reduced beta chain synthesis


+ Reduced production of chains

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0 complete absence of chains

Compensatory

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Globin chain synthesis

occurs in Thalassemias.


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Unbalanced production of

Globin chains in

Thalassemias causes

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Erythrocytes to be small,

hypochromic and sometimes

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deformed.

Blood Picture Of Thalassemia
There occurs intracellular

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accumulation of unmatched

Globin chains in the developing

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Erythrocytes

Precipitation of the Proteins,

which leads to cell destruction in

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the bone marrow.

Infective Erythropoiesis.
Mature functional RBC's

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do not reach the

peripheral blood to carry

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oxygen.
Types of Thalassemia

Thalassemia Minor

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Heterozygous State
Asymptomatic

Thalassemia Major

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Homozygous Type
Lethal at birth or in childhood.
Has many complications
Early and Continuous treatment

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of Thalassemia allows survival to

young adulthood .
-Thalassemia

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Suppression of Globin

genes

No/reduced globin chains

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synthesis.

Compensatory more /

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globin chains synthesized.

Abnormal Hb in -Thalassemia

vHbH ? 4

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vHb Bart - 4

?Affect normal function of Hb
?Anemia
?Fetal death

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Types Of Alpha Thalassemias

/ Normal

/-

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Mild microcytosis

/- -

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Mild microcytosis

-/-
-/- -

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Hemoglobin H disease

- -/- -

Hemoglobin Barts Disease ? Hydrops Fetalis

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Silent Carriers of

Thalassemia

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Out of 4 Gene there is missing

of only 1 Gene.

Remaining 3 genes produces

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sufficient chains for normal

Hb production.

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1-2 % of Hb Bart in cord blood.
Thalassemia Trait

2 Genes are deleted
Shows mild microcytic

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hypochromic anemia

Occasionally Hb H inclusions
Cord blood contains 2-10% of Hb Bart.

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Hb H inclusions can be seen

in RBC's after supra vital

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stain.

Cord blood contains 10-20%

Hb Bart.

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Hb H Disease

Type of Thalassemia where 3

genes absent

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Hb H present-Tetramer of chains.
Alters shape of RBC's
Shorten RBC life span.
Moderate hemolytic anemia.

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Hb Bart Disease

Most clinically severe form
Thalassemia.

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Where all 4 genes deleted
Total absence of chain

biosynthesis
Hb Bart major Hb found-

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tetramer of chains.

Hydrops Fetalis
(Fetal Anemia causes Edema)

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Hb Bart has extremely high

Oxygen affinity

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Allows Oxygen transport but no

release at tissues.

Hypoxia

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Still born infants/ die shortly

after birth.
Beta Thalassemia

--- Content provided by FirstRanker.com ---

Beta Thalassemia

Suppression of Globin gene.
Reduced/ no production of

--- Content provided by FirstRanker.com ---

beta globin chains.

Compensatory / Globin

chains biosynthesized.

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Abnormal Hb in beta Thalassemia:

HbF (22)

HbA2(22)

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Types Of Beta Thalassemia

Beta Thalassemia minor

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? heterozygous (or trait)

Beta Thalassemia major

- homozygous

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Beta Thalassemia Trait

No symptoms
Mild microcytic anemia

--- Content provided by FirstRanker.com ---

Beta Thalassemia Major

No beta chain produced

(no HbA)

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Cooley's Anemia
Homozygous disease
Beta Thalassemia Major

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Crippling disease of

childhood

Persistent HbF in age above 1

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year

Reduced unloading of oxygen at

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tissues.

Premature destruction of RBC's.
Severe hypochromic

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microcytic anemia occurs

gradually in the first year of life

Bone Marrow expansion

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Hypersplenism
Hepatosplenomegaly

MCV low
Severe Anemia

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Reticulocytosis
Extreme Poikilocytosis

(Different Shape ) and

--- Content provided by‌ FirstRanker.com ---

Anisocytosis (Different Size).
HPFH

Hereditary Persistence of

--- Content provided by‍ FirstRanker.com ---

Fetal Hb (HPFH)

It is a Genetic heterogeneous

disorder

--- Content provided by‍ FirstRanker.com ---


Caused due to deletions of

Genes in chromosome 11.

--- Content provided by​ FirstRanker.com ---

Exhibits total absence of

and Globin chain synthesis.

Hb F(2 2) is the

--- Content provided by‌ FirstRanker.com ---


predominant Hb present.
HPFH patients are

asymptomatic

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If they are sedentary

and slow workers.

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Diagnosis Of Thalassemia's

PCR based methods.

--- Content provided by‌ FirstRanker.com ---

Gene Mapping
For families that carry

a Thalassemia trait.

--- Content provided by​ FirstRanker.com ---

Genetic counseling

and genetic testing is

recommended

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Treatment Of Thalassemias

Repeated / frequent

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blood transfusions.

(After every 3- 4

months)

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Due to repeated blood transfusions

in patients of Beta Thalassemia.

There exhibits Iron overload

--- Content provided by‌ FirstRanker.com ---

Iron toxicity is noted since Iron is

one way element

Iron once entered in blood do not get

--- Content provided by​ FirstRanker.com ---


excreted out.

Iron gets accumulated in functional

--- Content provided by‍ FirstRanker.com ---

tissues.

Tissue dysfunctions,

Growth failure and death

--- Content provided by‌ FirstRanker.com ---


occurs before puberty due to

Iron toxicity.

--- Content provided by FirstRanker.com ---

However Iron chelation-

Reduces Iron toxicity.
Folate supplementation

--- Content provided by FirstRanker.com ---

for promotion of

Erythropoiesis

Azacytidine ? Drug used

--- Content provided by FirstRanker.com ---


with limited success

Bone marrow

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transplantation

Stem cell transplant
Gene therapy

--- Content provided by​ FirstRanker.com ---


Abnormal Hb Variants

With Increased Oxygen Affinity

--- Content provided by FirstRanker.com ---

Hb Bart
Hb H
Hb Chesapeake
Hb Rainier

--- Content provided by‍ FirstRanker.com ---


Catabolism/Breakdown Of Hb

OR

--- Content provided by​ FirstRanker.com ---

Formation and Fate Of Bilirubin

OR

How Bilirubin is Formed and

--- Content provided by⁠ FirstRanker.com ---


Excreted ?
Catabolism of Hemoglobin

begin after destruction of

--- Content provided by​ FirstRanker.com ---


RBC's.

RBC destruction is normally

--- Content provided by​ FirstRanker.com ---

the result of senescence

(Old/Aged).

Red cell destruction

--- Content provided by​ FirstRanker.com ---


usually occurs after a

mean life span of

--- Content provided by‍ FirstRanker.com ---

120 days.
The old red blood

cells are removed

--- Content provided by⁠ FirstRanker.com ---

Extravascularly by

Macrophages of R.E

System.

--- Content provided by‌ FirstRanker.com ---


Reticuloendothelial

system (RES), specially of

--- Content provided by​ FirstRanker.com ---

Spleen, Bone marrow and

Liver are involved in RBC

destruction.

--- Content provided by‌ FirstRanker.com ---

Essentials for Erythrocyte

Membrane Integrity

Continuous supply of Glucose to

--- Content provided by‍ FirstRanker.com ---


Erythrocytes

Continuous and uninterrupted

--- Content provided by‌ FirstRanker.com ---

Glycolysis in RBCs

Continuous minimal ATP

production in RBCs

--- Content provided by​ FirstRanker.com ---


RBC aging is characterized by:
Decreased Glycolytic enzyme

activity

--- Content provided by⁠ FirstRanker.com ---


Which leads to decreased

Glycolysis and ATP production

--- Content provided by‌ FirstRanker.com ---

Subsequent loss of deformability

and membrane integrity of RBCs.
Each day ~ 1% of the

--- Content provided by​ FirstRanker.com ---

RBCs are removed

and replaced.

Approximately 2- 3 million old

--- Content provided by⁠ FirstRanker.com ---


RBCs removed and same amt

of new red blood cells enter

--- Content provided by‌ FirstRanker.com ---

the circulation per second.

This maintains constant RBC

count in blood.

--- Content provided by‍ FirstRanker.com ---

80-90% of aged RBC destruction

is Extravascular

Occurs mainly in the

--- Content provided by FirstRanker.com ---


Macrophages, Mononuclear

phagocytic cells of Spleen

--- Content provided by‌ FirstRanker.com ---

Spleen is grave yard for RBC's

Small amount occurring in the

RE system of Liver and Bone

--- Content provided by​ FirstRanker.com ---


marrow.

10-20 % of RBC destruction

--- Content provided by FirstRanker.com ---

is Intravascular, occurring

in the lumen of the blood

vessels.

--- Content provided by‌ FirstRanker.com ---

Senescent /old RBC's

in RES are lysed to

release its contents-

--- Content provided by​ FirstRanker.com ---


Hemoglobin (Hb)

Hb is degraded to:

--- Content provided by​ FirstRanker.com ---

Globins Amino Acids

Recycled Metabolism
Heme Bilirubin

--- Content provided by‌ FirstRanker.com ---

excreted out of the body.

Fe2+ Transported as

Transferrin

--- Content provided by‍ FirstRanker.com ---


Iron stored as Ferritin

and reused in the next

--- Content provided by FirstRanker.com ---

Heme biosynthesis

Not only Hb but

other Hemoproteins

--- Content provided by⁠ FirstRanker.com ---


containing Heme

groups are degraded

--- Content provided by‍ FirstRanker.com ---

by the same pathway.


Extravascular Erythrocyte

--- Content provided by FirstRanker.com ---

Destruction

Extravascular Erythrocyte Destruction

is a normal pathway.

--- Content provided by⁠ FirstRanker.com ---


80-90% Erythrocytes destructed in

this manner.

--- Content provided by‍ FirstRanker.com ---

Outside the circulatory system.
Inside the phagocytic cells of Spleen,

Liver and Bone marrow.

--- Content provided by‍ FirstRanker.com ---


Extravascular destruction of

RBCs
Intravascular Erythrocyte

--- Content provided by⁠ FirstRanker.com ---


Destruction

Erythrocytes destructed in

--- Content provided by FirstRanker.com ---

circulatory system.

Normally 10 -20% erythrocytes

destructed in this manner.

--- Content provided by​ FirstRanker.com ---


Hb is directly released into blood

stream.

--- Content provided by‌ FirstRanker.com ---

Hb in blood is bound to

Haptoglobin.

Prevents renal excretion of

--- Content provided by​ FirstRanker.com ---


plasma Hb

Circulating Hb is removed

--- Content provided by‍ FirstRanker.com ---

by Liver.


Intravascular destruction of RBCs

--- Content provided by‍ FirstRanker.com ---


Globin chains are broken

down to amino acids

--- Content provided by FirstRanker.com ---


Which are reutilized for

general protein synthesis

--- Content provided by‍ FirstRanker.com ---

in the body.
Heme part is

catabolized to

--- Content provided by‍ FirstRanker.com ---

Bilirubin and

excreted out of the

body.

--- Content provided by‍ FirstRanker.com ---


Microsomal enzyme Heme

Oxygenase of RE cells acts on

--- Content provided by⁠ FirstRanker.com ---

Heme

Requires NADPH+H+ as a coenzyme
Cleaves Methenyl bridge of

--- Content provided by‍ FirstRanker.com ---

cyclic tetrapyrrole ring of Porphyrin

.

Forms Biliverdin ? A linear tetra

--- Content provided by FirstRanker.com ---


Pyrrole ring structure.
Iron is released in Ferrous is

oxidized to Ferric and

--- Content provided by‍ FirstRanker.com ---


transported by Transferrin.

CO released is expired out.

--- Content provided by‍ FirstRanker.com ---

Enzyme Biliverdin Reductase

Reduces Methenyl bridges of Biliverdin

to Methylene bridges.

--- Content provided by⁠ FirstRanker.com ---


Reduces Biliverdin (Green bile pigment )

to Bilirubin ( Yellow bile pigment).

--- Content provided by​ FirstRanker.com ---

NADPH+H+ is used as reducing equivalent

for this reduction reaction by Biliverdin

Reductase.

--- Content provided by FirstRanker.com ---

Albumin Transports Unconjugated

Bilirubin Through Blood.

1 gram of Hb yields 35 mg

--- Content provided by‌ FirstRanker.com ---


of Bilirubin.

Daily 250-300 mg of

--- Content provided by FirstRanker.com ---

Bilirubin is produced by an

adult.
Bilirubin formed in RE cells of a

--- Content provided by​ FirstRanker.com ---

Spleen after Heme catabolism and

released in the blood circulation is:

Non polar

--- Content provided by‍ FirstRanker.com ---

Insoluble
Free or Unconjugated Bilirubin

Albumin a polar moiety

--- Content provided by FirstRanker.com ---

helps in transporting this

non polar Bilirubin


--- Content provided by‌ FirstRanker.com ---

Through aqueous phase of

blood circulation up to

Liver.

--- Content provided by​ FirstRanker.com ---

Albumin has two binding

sites for Bilirubin.

?High affinity binding site

--- Content provided by⁠ FirstRanker.com ---

?Low affinity binding site

Bilirubin first tightly binds

to high affinity binding site

--- Content provided by⁠ FirstRanker.com ---


of Albumin.

25 mg of Bilirubin tightly

--- Content provided by​ FirstRanker.com ---

binds with Albumin in 100 ml

blood.
Bilirubin bound

--- Content provided by FirstRanker.com ---

with Albumin

Prevents urinary

excretion of

--- Content provided by‌ FirstRanker.com ---


Bilirubin in urine.

Drugs like Sulfonamides,

--- Content provided by⁠ FirstRanker.com ---

Penicillin, Salicylates

Compete with Bilirubin

for its binding to

--- Content provided by‍ FirstRanker.com ---


Albumin.
Hypoalbuminemia

affects transport

--- Content provided by‌ FirstRanker.com ---


and excretion of

Bilirubin.

--- Content provided by​ FirstRanker.com ---

Hypoalbuminemia may

lead to retention of

Unconjugated Bilirubin

--- Content provided by⁠ FirstRanker.com ---


in blood circulation.

May cause Bilirubin

--- Content provided by​ FirstRanker.com ---

Encephalopathy.
Facilitated transport

system helps in uptake of

--- Content provided by FirstRanker.com ---

Bilirubin in sinusoidal

surface of Hepatocytes.

Ligandin and Protein

--- Content provided by‌ FirstRanker.com ---


Y of Hepatocytes

Prevent efflux of

--- Content provided by​ FirstRanker.com ---

Bilirubin back into

blood stream.
Conjugation Of Bilirubin

--- Content provided by FirstRanker.com ---

In Liver

Non polar Bilirubin

entered in Liver

--- Content provided by‌ FirstRanker.com ---


Undergoes conjugation

reaction

--- Content provided by‌ FirstRanker.com ---

Conjugating agent is two

molecules of UDP-

Glucuronic acid

--- Content provided by‍ FirstRanker.com ---

In presence of conjugating

enzyme UDP Glucuronyl

Transferase

--- Content provided by‍ FirstRanker.com ---


Forms Conjugated Bilirubin-

Bilirubin Diglucuronide.

--- Content provided by​ FirstRanker.com ---

Conjugated Bilirubin is:

Polar
Soluble form
Readily excretable form.

--- Content provided by‍ FirstRanker.com ---

Conjugated Bilirubin is carried

through Bile via common bile duct

and excreted in the intestine

--- Content provided by​ FirstRanker.com ---



Secretion of Bilirubin into the

bile occurs by an active

--- Content provided by⁠ FirstRanker.com ---


transport mechanism.

Conjugated Bilirubin reaches terminal

--- Content provided by‌ FirstRanker.com ---

ileum and large intestine.

Glucuronides are removed by specific

bacterial enzymes- Glucuronidase.

--- Content provided by‌ FirstRanker.com ---


Bilirubin is reduced to colorless

compound Stercobilinogen in

--- Content provided by‌ FirstRanker.com ---

intestine.
Small amount of Stercobilinogen is

reabsorbed and re excreted through

--- Content provided by‍ FirstRanker.com ---

the Liver ? Enterohepatic

circulation.

Stercobilinogen is partly reabsorbed

--- Content provided by​ FirstRanker.com ---


enters in blood circulation is excreted

in urine as Urobilinogen and Urobilin.

--- Content provided by‌ FirstRanker.com ---

Most of the

Stercobilinogen of

intestine is oxidized to

--- Content provided by⁠ FirstRanker.com ---


Stercobilin a orange

yellow colored

--- Content provided by‌ FirstRanker.com ---

compound.
Stercobilin is a

major excretable

--- Content provided by⁠ FirstRanker.com ---

form of Bilirubin in

feces.

Futher Fate of Bilirubin

--- Content provided by⁠ FirstRanker.com ---


Bilirubin (Bil) is released from RES into the blood.
BUT! Bil is only poorly soluble in plasma, and therefore

during transport it is bound to albumin (Unconjugated

--- Content provided by⁠ FirstRanker.com ---


Bilirubin").



--- Content provided by‌ FirstRanker.com ---

LIVER
In the hepatocytes, Bil is conjugated by 2 molecules of

glucuronic acid bilirubin diglucuronide (soluble in

--- Content provided by‍ FirstRanker.com ---

water, ,,conjugated Bil")



BILE

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INTESTINE
Bilirubin is reduced to urobilinogen and stercobilinogen

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BLOOD

CELLS

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Stercobilin

excreted in

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Urobilin

Hemoglobin

feces

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excreted in

urine

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Globin

Urobilinogen

Heme

--- Content provided by‍ FirstRanker.com ---


formed by bacteria

KIDNEY

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O2

reabsorbed

Heme

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INTESTINE

into blood

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oxCyO

genase

Biliverdin IX

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via bile duct to intestines

NADPH

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Biliverdin

Bilirubin diglucuronide

reductase

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(water-soluble)

NADP+

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2 UDP-glucuronic acid

Bilirubin

Bilirubin

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LIVER

(water-

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(water-insoluble)

via blood

insoluble)

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to the

liver

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Catabolism of Hemoglobin


Normal Red Cell Breakdown

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Hemoglobin

Heme

Globin

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Iron

Protoporphyrin

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Amino acids

CO

Bilirubin

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Transferrin

Expired air

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(free)

Liver

conjugation

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Erythroblast

Bilirubin Glucuronides

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Urobilin(ogen)

Stercobilin(ogen)

Urine

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Feces


Red Blood Cell Turnover

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What Is Bilirubin ?

Bilirubin is

Metabolic waste, an end product of Heme

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catabolism

Formed in cells of RE system mainly in Spleen

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Richly present in Bile

Yellow colored Bile pigment

Carried through Bile for its excretion through

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feces.
Recently a research

study has depicted

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Bilirubin has an

Antioxidant capacity.

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Bilirubin is majorly

excreted out through

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feces in the form of

Stercobilin.

(Yellow Orange pigment)

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Types Of Bilirubin

Free/ Unconjugated /Indirect

Bilirubin

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Non polar / insoluble form

Formed in RES- Spleen ,

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Liver

Present in blood circulation

before entry into Liver cells.

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Bilirubin Diglucuronide /Conjugated/

Direct Bilirubin

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Polar/ Soluble form of

Bilirubin

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Unconjugated Bilirubin is

transformed to conjugated

Bilirubin in Liver.

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Bilirubin-diglucuronide = Conjugated Bilirubin

is soluble in water "Direct Bilirubin"
Conjugated Bilirubin is formed

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after conjugation reaction,

Conjugated with Glucuronate
Conjugated Bilirubin is

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readily mixed with bile and

excreted out through feces.

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Normal Levels

Serum Bilirubin

S.

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Type Of Bilirubin

Normal

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No

Ranges

1

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Total Bilirubin

0.2- 1 mg %

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Direct+ Indirect

2 Unconjugated/Indirect 0.2-0.8 mg%

Bilirubin

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3

Conjugated / Direct 0- 0.2 mg%

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Bilirubin
In normal healthy

conditions there is

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No conjugated

Bilirubin in

circulating blood.

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When Is Conjugated Bilirubin

is present in blood?

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During obstruction in common

bile duct (CBD)

(Obstructive Jaundice)

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Forms of Bilirubin

Bilirubin - In Bile
Urobilin ? In Urine
Stercobilin - In Stool

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Hyperbilirubinemia
Hyperbilirubinemia

Increased levels of serum

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Total Bilirubin

Above 1 mg% is termed

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as Hyperbilirubinemia.

Types Of Hyperbilirubinemia

Unconjugated

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Hyperbilirubinemia

Conjugated

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Hyperbilirubinemia

Biphasic Hyperbilirubinemia.
Causes of Hyperbilirubinemia

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Over production Of Bilirubin

Less excretion of Bilirubin

Leads to retention of Bilirubin in

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blood causing

Hyperbilirubinemia.
Causes of Hyperbilirubinemia

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Hemolytic anemia

Hepatitis

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Biliary duct stone

excess

hemolysis

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Unconjugated Bilirubin

Unconjugated

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conjugated Bilirubin

(in blood)

Bilirubin

--- Content provided by‍ FirstRanker.com ---


(in blood)

conjugated Bilirubin

--- Content provided by‌ FirstRanker.com ---

(in blood)

(released to bile duct)

conjugated Bilirubin

--- Content provided by FirstRanker.com ---


(in blood)

Causes of Hyperbilirubinemia

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1.Conditions which

form excess Bilirubin

than the excreting

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capacity of body.
Abnormal excessive intravascular

hemolysis

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Overproduction of Bilirubin from Heme

catabolism.

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More than the capacity of Liver to

conjugate and excrete

Leads to Unconjugated

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Hyperbilirubinemia.

2.Conditions which

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block excretion of

Bilirubin out of the

body.

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Failure of diseased Liver to

conjugate and excrete

Bilirubin through bile.

--- Content provided by‌ FirstRanker.com ---


In viral hepatitis
Leads to biphasic

Hyperbilirubinemia.

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Obstruction to the flow of bile
Due to obstruction in bile duct.
Regurgitation of bile in systemic

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blood circulation.

Leads to conjugated

Hyperbilirubinemia.

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3.Congenital / Inherited

defects in uptake and

excretion of Bilirubin by

--- Content provided by​ FirstRanker.com ---


Liver
Leads to congenital

Hyperbilirubinemia.

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4.Hypoalbuminemia / high

drug concentration in blood
Affects Bilirubin transportation

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by Albumin

Retains Bilirubin in blood
Leads to Unconjugated

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Hyperbilirubinemia.
Diagnosis of Hyperbilirubinemia

Diagnosis of Hyperbilirubinemia

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Vanden Bergh's Reaction
Quantitative estimation of

serum:

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Total Bilirubin
Unconjugated Bilirubin
Conjugated Bilirubin
Types of Vanden Bergh's Reaction

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Direct Vanden Bergh's Reaction

Estimates serum Conjugated

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Bilirubin (soluble form)

Serum Conjugated Bilirubin +

Diazo Reagent = Pink Azobilirubin

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In the Direct Vanden Bergh

Reaction soluble form of a Bilirubin

is directly and immediately

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reacted with the Diazo Reagent.

Conjugated Bilirubin requires no

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solubilizing agent.

Hence Conjugated Bilirubin is

also termed as Direct Bilirubin

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Indirect Vanden Bergh's Reaction

Estimates Serum Unconjugated

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Bilirubin (Insoluble form)

Serum Unconjugated Bilirubin +

Methanol/Surfactant (solubilizing

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agent)+ Diazo Reagent = Pink

Azobilirubin
In an Indirect Vanden Bergh

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Reaction

Insoluble form of Unconjugated

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Bilirubin is first solubilized with a

solubilizing agent (Methanol)

Then the solubilized form reacts with

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Diazo reagent to form a pink

Azobilirubin complex

--- Content provided by‍ FirstRanker.com ---

Unconjugated Bilirubin indirectly

reacts with the Diazo Reagent.

Since Unconjugated Bilirubin

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requires solubilizing agent for

reaction with Diazo reagent.

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Hence Unconjugated Bilirubin is

also termed as Indirect Bilirubin
Results and Significance

--- Content provided by FirstRanker.com ---

Of

Vanden Bergh's Reaction

S.No

--- Content provided by FirstRanker.com ---


Results of

Type of Hyperbilirubinemia/

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Vanden Bergh

Jaundice

1

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Direct Vanden Conjugated Hyperbilirubinemia

Bergh's Reaction

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Obstructive Jaundice

Positive

2

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Indirect Vanden

Unconjugated

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Bergh's Reaction

Hyperbilirubinemia.

Positive

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Hemolytic Jaundice

3

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Both Direct and Biphasic Hyperbilirubinemia

Indirect Vanden

means

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Bergh's Reaction

Both conjugated and

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positive

Unconjugated Bilirubin

increased.

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Hepatic Jaundice.
Significance Of

Vanden Bergh's Reaction

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I. Quantitatively Estimates serum

Total, conjugated and

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Unconjugated Bilirubin.

II. From the levels of serum total

Bilirubin- Diagnoses Jaundice

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III. From the serum levels of Direct and

Indirect Bilirubin levels-

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Differentiate- Type of Jaundice

IV. From the values of serum Bilirubin-

Indicate-Severity of Jaundice

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Jaundice/Icterus Condition

Jaundice is a

pathological/Clinical

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condition

Characterized by

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Hyperbilirubinemia
In Jaundice Total

Serum Bilirubin levels

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are more than 2.5 mg %.

High circulating

Bilirubin more than

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2.5 mg% in blood

and tissues causes

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jaundice.


Jaundice is a yellow

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discoloration to:

Skin
Sclera of eyes
Nails

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Mucous membrane
Causes and Types Of Jaundice

Basic Cause and Consequence

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Of Jaundice

Defect in Heme catabolism

vOverproduction of Bilirubin

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Defect in Bile excretion

?Less excretion of Bilirubin

--- Content provided by⁠ FirstRanker.com ---

Retains Bilirubin in blood and

body


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Jaundice

Types Of Jaundice
Hemolytic/Pre Hepatic /Retention/

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Acholuric Jaundice

Hepatic/Infectitious Jaundice

Obstructive/Post Hepatic/

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Regurgitation /Choluric/Cholestatic

Jaundice.

Neonatal Physiological Jaundice

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Neonatal Pathological Jaundice

Erythroblastosis Foetalis

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Breast Feeding Jaundice

Breast Milk Jaundice
Sub Clinical Jaundice/ Latent

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Jaundice-

Serum Bilirubin levels between 1-3

mg%

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Clinical Jaundice-

Serum Bilirubin levels more than

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3 mg%.

Hemolytic/ Prehepatic

/Retention Jaundice

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Due to abnormal excessive

intravascular

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hemolysis(premature) .

Characterized by Unconjugated

Hyperbilirubinemia.

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Healthy Body RBC Destruction

Normal human body brings approx.

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200 billion RBC lysis per day

v 160 billion is Extravascular Lysis
v 40 billion is Intravascular lysis

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Intravascular destruction of RBCs
Conditions Causing Abnormal

Excessive Intravascular Hemolysis

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Sickle Cell Anemia
Thalassemia's
Glucose-6-Phosphate Dehydrogenase

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deficiency

Mismatched blood transfusion
Malaria
Burns

--- Content provided by​ FirstRanker.com ---

Spherocytosis
Drug interactions
Excessive abnormal intravascular

hemolysis

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Increased Heme catabolism
Increased Unconjugated Bilirubin

in blood

--- Content provided by​ FirstRanker.com ---


Levels of the Unconjugated Bilirubin

more than normal capacity of Liver

--- Content provided by‌ FirstRanker.com ---

to conjugate and excrete (3 gm/day).

Unconjugated Bilirubin is in

queue to enter Liver for

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conjugation and excretion.

Thus there occurs retention of

--- Content provided by FirstRanker.com ---

Unconjugated Bilirubin in

blood.
Laboratory Findings

--- Content provided by‍ FirstRanker.com ---

In

Hemolytic Jaundice.

Blood Investigations

--- Content provided by‍ FirstRanker.com ---


Results of Vanden Bergh's

Reaction

--- Content provided by​ FirstRanker.com ---

Total Bilirubin Increased
Indirect Bilirubin Increased
Direct Bilirubin Normal
Urine Investigations

--- Content provided by‍ FirstRanker.com ---

Ehrlich's Test for urine

Urobilinogen ? Positive

Urine Urobilin increased.

--- Content provided by​ FirstRanker.com ---

Hays sulfur test for Bile salts-

Negative

Fouchet's Test for Bilirubin-

--- Content provided by FirstRanker.com ---


Negative

Stool Appearance

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Dark brown color feces in

Hemolytic Jaundice

Due to more Stercobilin

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excreted out in feces.
Bone Marrow Examination

Hyperplasia of Bone marrow

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Reticulocytosis in Peripheral

blood smear

Immature form of RBC's

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increased in blood circulation.

Hepatic/ Infectitious Jaundice

--- Content provided by‌ FirstRanker.com ---

Liver Parenchymal damage
Due to Viral Hepatitis
Biphasic

Hyperbilirubinemia.

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Conditions Affecting Liver

Parenchymal Damage

Viral infection of Liver

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( Viral Hepatitis)
Liver Cirrhosis- Alcoholism
Drug Effects:

vRifampicin -affect cellular

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uptake of Bilirubin by Liver cells

vNovobiocin -affect conjugation

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of Bilirubin in Liver.
Causes Of Biphasic

Hyperbilirubinemia in Hepatic

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Jaundice

In Hepatitis damage and

inflammation of Liver parenchymal

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cells.

This impairs and delays the

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conjugation and excretion of Bilirubin

by Liver.

This retains Unconjugated Bilirubin

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in blood.
Inflammation of Hepatocytes

and intra hepatic obstruction

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in hepatitis

Leaks out conjugated

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Bilirubin in blood.

Thus biphasic

Hyperbilirubinemia is noted.

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In Hepatic Jaundice

there is a Marginal

--- Content provided by​ FirstRanker.com ---

increase

In both serum

Unconjugated and

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Conjugated Bilirubin
Laboratory Findings

In

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Hepatic Jaundice

Blood Investigations

--- Content provided by FirstRanker.com ---

Results of Vanden Bergh's Reaction

Total Bilirubin increased
Indirect Bilirubin increased (Marginal)
Direct Bilirubin increased (Marginal)

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Serum SGPT/ALT and SGOT/AST

activity increased.
Urine Investigations

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Ehrlich's Test for urine

Urobilinogen ? Normal

--- Content provided by FirstRanker.com ---

Urine Urobilin normal.
Hays sulfur test for Bile salts-

may be positive in severe cases.

--- Content provided by​ FirstRanker.com ---

Fouchet's Test for Bilirubin-may

be positive in severe cases.

Stool Appearance

--- Content provided by FirstRanker.com ---


Normal/ slightly pale colored

feces in Hepatic Jaundice.
Obstructive/ Post hepatic /

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Regurgitative Jaundice

Due to obstruction in bile

--- Content provided by​ FirstRanker.com ---

flow to reach small intestine.

Characterized with

Conjugated

--- Content provided by FirstRanker.com ---


Hyperbilirubinemia.

Conditions Causing Obstructive

--- Content provided by‌ FirstRanker.com ---

Jaundice
Obstruction of Bile duct due to

Gall stones in Common Bile

--- Content provided by FirstRanker.com ---

Duct(CBD).

Narrowing of bile duct due to

surgery.

--- Content provided by‍ FirstRanker.com ---


Tumor of head of Pancreas.
Enlargement of lymph glands

near Gall bladder /bile duct.

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Causes Of Conjugated

Hyperbilirubinemia In Obstructive

--- Content provided by​ FirstRanker.com ---

Jaundice.
Partial/ Complete Obstruction of

bile duct

--- Content provided by‌ FirstRanker.com ---

Regurgitation of bile into systemic

circulation

Bile contains bile salts and bile

--- Content provided by‌ FirstRanker.com ---


pigment- Bilirubin .

Hydrophilic Conjugated Bilirubin

--- Content provided by FirstRanker.com ---

is now in blood circulation.

In Obstructive Jaundice

Bile get excreted out through urine.

--- Content provided by‌ FirstRanker.com ---

Bile salts and conjugated Bilirubin

present in urine.

Dark yellow colored urine noted

--- Content provided by​ FirstRanker.com ---


due to presence of Bilirubin occurs in

Obstructive Jaundice patients.
Laboratory Findings

--- Content provided by FirstRanker.com ---


In

Obstructive Jaundice.

--- Content provided by‍ FirstRanker.com ---

Blood Investigations

Results of Vanden Bergh's

Reaction

--- Content provided by​ FirstRanker.com ---


Total Bilirubin increased
Indirect Bilirubin normal
Direct Bilirubin increased
Serum ALP activity increased.

--- Content provided by​ FirstRanker.com ---

Urine Investigations

Ehrlich's Test for urine

Urobilinogen ? Negative

--- Content provided by FirstRanker.com ---


Urine Urobilin decreased.
Hays sulfur test for Bile salts-

Significantly Positive

--- Content provided by FirstRanker.com ---


Fouchet's Test for Bilirubin-

Significantly Positive

--- Content provided by‌ FirstRanker.com ---

Stool Appearance

Clay colored stools due to

absence of Stercobilin in feces of

--- Content provided by⁠ FirstRanker.com ---


Obstructive Jaundice.

Fatty stools due to excretion of

--- Content provided by FirstRanker.com ---

Lipids in feces

(Absence of Bile salts in intestine).
Neonatal

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Physiological Jaundice

Neonatal - Physiological Jaundice

Noted in premature, low

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birth weight infants.

After 1- 7 days of birth.
Causes

--- Content provided by FirstRanker.com ---


Immature hepatic system

in premature born infants

--- Content provided by FirstRanker.com ---

Poor uptake and

conjugation of

Unconjugated Bilirubin

--- Content provided by‌ FirstRanker.com ---


from blood by Liver

Low levels of Conjugating Enzyme
UDP- Glucuronyl Transferase

--- Content provided by FirstRanker.com ---

Delays the conjugation and

excretion of Bilirubin
Consequences

--- Content provided by FirstRanker.com ---

Physiological Jaundice exhibits

Unconjugated

Hyperbilirubinemia

--- Content provided by‌ FirstRanker.com ---


Serum Bilirubin may raise up to

20 mg % or more

--- Content provided by​ FirstRanker.com ---

Unconjugated Bilirubin is

hydrophobic

It can easily cross blood brain

--- Content provided by FirstRanker.com ---


barrier to enter central nervous

system.

--- Content provided by FirstRanker.com ---

Leading to Kernicterus

(Bilirubin Encephalopathy)
Bilirubin accumulates in

--- Content provided by⁠ FirstRanker.com ---

Neurons of Basal Ganglia,

Hippocampus Cerebellum
Medulla of Brain.
This causes necrosis of

--- Content provided by⁠ FirstRanker.com ---


nerve cells and brain

damage.

--- Content provided by FirstRanker.com ---

Symptoms

Fits/ Convulsions
Mental Retardation
Encephalitis

--- Content provided by FirstRanker.com ---

Spasticity
(Skeletal muscle tightness and

stiffness)
Treatment

--- Content provided by FirstRanker.com ---


Phototherapy at 450 nm.

Exchange transfusions of blood
(When Serum Bilirubin > 20 mg%)

--- Content provided by​ FirstRanker.com ---


During Phototherapy baby is

exposed to uv light

--- Content provided by‌ FirstRanker.com ---

The Bilirubin is transformed to

solubilized form of Bilirubin


--- Content provided by​ FirstRanker.com ---

Which is readily excretable out

through urine.
?Blue / white UV light induces

--- Content provided by⁠ FirstRanker.com ---

isomerization

?Of non polar , insoluble form of

Unconjugated Bilirubin ?Z isomer

--- Content provided by⁠ FirstRanker.com ---


?To water soluble, polar form of

Bilirubin- E isomer.

--- Content provided by​ FirstRanker.com ---

The phototherapy

should be exposed to

child's skin

--- Content provided by‍ FirstRanker.com ---


Breast feed the child

every 2 to 3 hours

--- Content provided by‍ FirstRanker.com ---

(10 to 12 times a day).
Feeding prevents

dehydration and helps Bilirubin

--- Content provided by‌ FirstRanker.com ---

to excrete out the body.

Phototherapy will continue until

the baby's serum Bilirubin

--- Content provided by⁠ FirstRanker.com ---


levels are low enough to be

safe.

--- Content provided by FirstRanker.com ---

Neonatal-Pathological Jaundice
Erythroblastosis Foetalis

Hemolytic condition in

--- Content provided by⁠ FirstRanker.com ---

neonates

Extrinsic Cause of Hemolysis

Exhibits Unconjugated Hyper

--- Content provided by​ FirstRanker.com ---


Bilirubinemia

Cause

--- Content provided by‌ FirstRanker.com ---

Caused due to Rh incompatibility

When Rh ? ve mother

conceives Rh + ve baby.

--- Content provided by‍ FirstRanker.com ---


This causes excessive

hemolysis of Erythrocytes at

--- Content provided by​ FirstRanker.com ---

the time of birth.
Neonatal

Pathological Jaundice

--- Content provided by FirstRanker.com ---

Non-Hemolytic Cause

Breast Feeding Failure

Jaundice

--- Content provided by‍ FirstRanker.com ---


Jaundice caused in new

born infants.

--- Content provided by​ FirstRanker.com ---

Due to insufficient/lack

in breast feeding of milk
Infants born by cesarean

--- Content provided by‌ FirstRanker.com ---

section are at higher risk

for this condition.

Due to no normal

--- Content provided by FirstRanker.com ---


Lactation phase.

Inadequate quantities of

--- Content provided by⁠ FirstRanker.com ---

milk reached to infants

body.

Decreases body fluids

--- Content provided by‍ FirstRanker.com ---


,lowers bowel movements

,impair to remove

--- Content provided by⁠ FirstRanker.com ---

Bilirubin from an infants

body.
Proper feeding

--- Content provided by FirstRanker.com ---

prevents dehydration

Helps Bilirubin to

excrete out of the body

--- Content provided by‍ FirstRanker.com ---


without its retention.

Condition of breast feeding

--- Content provided by⁠ FirstRanker.com ---

Jaundice can be ameliorated by

Frequent breast feeding

sessions of sufficient

--- Content provided by‌ FirstRanker.com ---


duration(10-12 /day)

This stimulate adequate milk

--- Content provided by‍ FirstRanker.com ---

production by mothers breast.
Extra fluids are helpful for babies who

have not been getting enough breast

--- Content provided by FirstRanker.com ---

milk.

Nursing more often (up to 12 times a day)
Will increase the baby's fluid levels

--- Content provided by​ FirstRanker.com ---

Can cause the Bilirubin level to drop.


Breast Milk Jaundice
Breast Milk Jaundice

--- Content provided by⁠ FirstRanker.com ---


Non Organic Cause

Breast milk Jaundice is more of a

--- Content provided by⁠ FirstRanker.com ---

biochemical problem

Probably caused by

factors/chemicals present in the

--- Content provided by FirstRanker.com ---


Breast milk.

These may block certain Proteins/

--- Content provided by FirstRanker.com ---

Enzymes in the infant Liver that

metabolize Bilirubin.

Breast Milk Jaundice tends to

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run in families

It occurs equally often in Males

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and Females

Affects 0.5 % to 2.4% of all

newborns.

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Hypothesized Mechanisms

Increased levels of Epidermal

Growth Factor (EGF) in Breast

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milk.

Increased Bilirubin uptake from

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the gut (enterohepatic

circulation) in breast fed babies.

In a new born Liver, activity of

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Glucuronyl Transferase is only at 0.1-

1% of adult levels

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Conjugation of Bilirubin in infants is

reduced in comparison to adults.

Further inhibition of Bilirubin

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conjugation by other agents leads

to increased levels of Bilirubin in the

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blood.
Breast-milk of some women contains a

metabolite of Progesterone called

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3-Alpha-20-beta Pregnanediol.

This metabolite inhibits the action of

the conjugating enzyme Uridine Di

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Phospho (UDPGA) Glucuronyl

Transferase.

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This brings poor conjugation and

subsequent excretion of Bilirubin.

An enzyme in breast milk called

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Lipoprotein Lipase produces increased

concentration of non esterified free

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fatty acids

That inhibit Hepatic UDP Glucuronyl

Transferase

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Which again leads to decreased

conjugation and subsequent excretion of

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Bilirubin.
Mothers taking drugs like Novobiocin,

Steroidal derivatives or Rifampicin

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Drugs secreted through breast milk
Infant fed by this milk has drug inhibitory

effect on Bilirubin metabolism.

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Delay in Bilirubin uptake and

conjugation in infants Liver.

Leads to Unconjugated Hyperbilirubinemia.

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Management Of

Breast Milk Jaundice

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Temporary stoppage of breast

milk feeding

Till the drug is cleared away

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from the breast fed milk.
Congenital Hyperbilirubinemia

Congenital Hyperbilirubinemia

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Genetic defects in Bilirubin

uptake, conjugation and

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excretion of Bilirubin

Leads to elevated levels of

Bilirubin in infants blood and

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body tissues.
Congenital Defect

Type Of

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Disorders

Hyperbilirubinemia

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Gilbert's

Defect in

Unconjugated

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Syndrome

uptake

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Hyperbilirubinemia

of

Bilirubin

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by Liver

cells

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Congenital Defect

Type Of

Disorders

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Hyperbilirubinemia

Crigler

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Complete

Unconjugated

Najjar

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absence of

Hyperbilirubinemia

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Syndrome-I

enzyme

UDP-

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Glucuronyl

Transferase
Congenital Defect

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Type Of

Disorders

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Hyperbilirubinemia

Crigler

Partial

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Unconjugated

Najjar

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absence of

Hyperbilirubinemia

Syndrome-

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enzyme

II

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UDP-

Glucuronyl

Transferase

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Congenital Defect

Type Of

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Disorders

Hyperbilirubinemia

Dubin

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Defect in

Conjugated

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Johnson's Hepatic

Hyperbilirubinemia

Syndrome/ excretion

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of

Deposition of Bilirubin

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Black Liver conjugated

in Liver

Jaundice

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Bilirubin
Congenital Defect

Type Of

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Disorders

Hyperbilirubinemia

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Rotors

Unknown

Conjugated

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Syndrome Autosomal

Hyperbilirubinemia

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Recessive

Inheritance Deposition of Bilirubin

in Liver

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Conditions Causing

Unconjugated Hyperbilirubinemia
Conditions Causing

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Unconjugated Hyperbilirubinemia

Hemolytic Jaundice
Hepatic Jaundice

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Neonatal/ Physiological Jaundice
Breast Milk Jaundice
Gilbert's Syndrome
Crigler Najjar Syndrome
Hypoalbuminemia

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High Drug Concentration lowering Albumin

activity.

Conditions Causing Conjugated

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Hyperbilirubinemia

Obstructive Jaundice
Hepatic Jaundice

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Dubin Johnson's Syndrome
Rotors Syndrome
Differential Diagnosis Of Jaundice

Parameters

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Hemolytic

Hepatic

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Obstructive

Jaundice

Jaundice

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Jaundice

Serum Bilirubin Indirect Bilirubin/

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Biphasic

Direct Bilirubin/

Unconjugated

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Both Direct

Conjugated

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Bilirubin

and

Bilirubin

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increased

Indirect

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Increased

Bilirubin

Increased

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Urine

Increased

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Normal or

Absent or

Urobilinogen

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Decreased

Decreased

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Urine Bilirubin

Absent

May present in

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Present in high

and Bile Salts

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small amounts

amounts

Fecal Stercobilin

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Increased

Normal or

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In traces or

Color Of Feces

Dark color feces

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Decreased

absent

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Clay colored

feces

Serum ALT

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Normal

Significantly

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May be slightly

activity

Increased

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increased

Serum ALP

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Normal

May slightly

Significantly

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activity

increased

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Increased
Questions

SHORT NOTES

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Outline of Heme Biosynthesis
Porphyrias: Types , Causes

and Consequences

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Acute Intermittent Porphyria

(AIP).
Catabolism of Hb /Fate &

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Formation of Bilirubin.

Jaundice & its types.
Vanden-Berg's Reaction

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and its significance.

Name the enzymes of Heme

Biosynthesis inhibited during

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lead poisoning. What is its

consequence?

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Comparison between abnormal

Hb derivatives & abnormal Hb

variants.

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Abnormal Hb variants
Hemoglobinopathies
Sickle Cell Anemia
Thalassemias
Thalassemia

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Thalassemia

Name the Bile pigments &

give the significance of their

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presence in blood & urine.

Laboratory reports of

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Obstructive Jaundice.

When and Why Bilirubin

comes in Urine Explain?

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Neonatal Jaundice
Breast Milk and Breast

Feeding Jaundice

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Congenital

Hyperbilirubinemias

Write the defect & type of

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Hyperbilirubinemia in

Crigler's Najjar Syndrome-I
Crigler's Najjar Syndrome-II

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Gilbert's Disease.
Dubin Johnson Syndrome.


THANK YOU

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