Agus Surono

October 2010

Cytogenetic
Genetics
Diagnostic
Engineering
Gene therapy

Kuliah Biomol 2010

Kuliah Biomol 2010

Human Genome Projects

Kuliah Biomol 2010

Regulation of gene :
DNA

Transcription

Nucleus

mRNA

Cytoplasm
Translation
Protein

Intracellular
Membran cell
Extracellular

Kuliah Biomol 2010

Protein Synthesis
Nucleus

Cytoplasm

transcription

DNA
Kuliah Biomol 2010

RNA

translation

Protein
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Protein maturation
Postranslational modification
- glycosilation
- phosphorylation
Folding

Resite
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Origin

Copy ????
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Identification/Diagnostic
Autosomal Dominant
Autosomal Recessive
X-Linked
Multifactorial

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The haploid human genome has

3 X 109 base pairs of DNA
sequence

This contains approximately

50,000 - 100,000 genes (more?)

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FISH: Fluorescence In Situ Hybridization

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Single Copy DNA (60%)

Mainly Euchromatic regions
Genes, Pseudogenes

Repetitive DNA (40%)

Mainly Heterochromatic regions
Tandem repeats
Microsatellite repeats
Minisatellite repeats

Interspersed repeats
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Identify large families with the

disease
Clinical Assessment.

Identify a chromosomal locus

Linkage.

Refine the candidate interval

Run more markers, construct a contig.

Identify Candidate Genes

Databases, exon trapping, direct selection.

Mutation Analysis
Sequencing, southern blotting, SSCP, ....
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INTRODUCTION OF
HEMATOLOGY

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Cellular Components
Components of Blood

Red Blood Cells

White Blood Cells
Inflammatory process
Coagulation

Hemorrhage
Blood Typing
Clotting disorders
Transfusion Medicine

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Spleen
Spleen

Blood
Blood

Kidneys
Kidneys
Hematopoietic
Hematopoietic
System
System

Bone
Bone
Marrow
Marrow
Kuliah Biomol 2010

Liver
Liver

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Pluripotent Stem Cell

Myeloid Multipotent
Stem Cells

Common Lymphoid
Stem Cells

Unipotent
Progenitors

Lymphocytes

WBCs
Basophils
Eosinophils
Neutrophils
Monocytes

RBCs
Thrombocytes

Platelets
Kuliah Biomol 2010

Erythrocytes
Erythropoietin

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Plasma
Transport
mechanism
90-92% water.
6-7% proteins
2-3%
Fats
Carbohydrates
(glucose)
Electrolytes
Gases (O2, CO2)
Chemical
messengers
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Components of Blood
Red Blood Cells
Erythrocyte
Hemoglobin O2
bearing molecule
Comprised of 4
subunits:
Globin (binds to 1 O2
molecule)
Heme (iron)
100% saturation = 4
globin subunits
carrying O2
Each gram of
hemoglobin = 1.34
ml O2
Kuliah Biomol 2010

Cellsalive.com

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Components of Blood
Red blood cell
production:
Erythropoiesis
Erythropoietin

Hemolysis
Sequestration
Laboratory analysis of
red blood cells
Red blood cell count
Hematocrit
Hemoglobin
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Components of Blood

Platelets
(Thrombocytes)
Megakaryocytes
Thrombopoietin

Thrombocytopenia
Thrombocytosis

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Class I

Class II

Class III

Blood Loss (ml)

Up to 750

750-1500

1500-2000

>2000

Blood Loss (%
volume)

Up to
15%

15%-30%

30%-40%

>40%

Pulse

<100

>100

>120

>140

BP

Normal

Normal

Decreased

Decreased

Pulse Pressure

Normal or Decrease
increased d

Decreased

Decreased

Resp. Rate

14-20

20-30

30-40

>35

Urine Output
(ml/hr)

>30

20-30

5-15

Negligible

Mental Status

Slightly
anxious

Mildly
anxious

Anxious,
confused

Confused,
lethargic

Fluid
Replacement
(3:1) rule

Crystalloi
d

Crystalloi
d

Crystalloid
and blood

Crystalloid
and blood

Kuliah Biomol 2010

Class IV

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Blood Types
Antigens
A, B, AB, O

Rh factor
Rh+ = ~85%
Rh- = ~15%

Blood transfusion
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Blood Typing - ABO

Blood type
Antigen
Antibody
present on
present in
erythrocyte
serum
O
AB
B
A

None
A and B
B
A
Kuliah Biomol 2010

Anti-A, Anti-B
None
Anti-A
Anti-B
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Blood Products and Blood Typing

Compatibility Among ABO Blood Groups
Reaction with serum of Recipient
Donor Cells

AB
B
A
O

AB
-

B
+
+
-

A
+
+
-

O
+
+
+
-

-= no reaction
+ = reaction
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Blood Products and Blood Typing

Brady; Paramedic Care Principles and Practice

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Membrane defect of
erythrocytes:

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The Red Cell

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SINTESIS Hb & HEMOLISIS

Eritrosit

Hb
300 juta/sel

Heme + Globin

non

Fe + Porfirin

Chain +
(,,)

bilirubin

Membrane defect of
erythrocytes:
Bilayer lipid membrane:
- Protein,

lipid, karbohidrat (50,40,10%)

- Phospholipid
- Attach to cytoskeleton

Strength, flexibility (deformability) erythrocytes

Protein (integral, peripheral)

Protein transmembrane
- Band 3 (ABO, I, i)
- Glycophorine A
C
- Protein Rh

(integral)

(M, N, P. falciparum),

Function: transport, pump, signal

transduction

(S, s, U),

Protein cytoskeleton

(peripheral)

Spectrin , (band 1,2), actin, ankyrin,

band 4.1, band 4.2
Cell stability function:
vertikal : spectrin, band 3, ankyrin, protein 4.2
horizontal: spectrin, actin, protein 4.1

Membrane defect of
erythrocytes:
Hereditary Spherocytosis
Pathophysiology:
Defect spectrin
Permeability Na
K, dehidration

Cell membrane
is not well
constructed

LIEN
conditioning effect
Sferis, fragil
Destruction
M, fisik

HEMOLISIS

Membrane defect of
erythrocytes:
Gene defect protein defect Clinically:
hemolytic
Band

Protei

BM

/sel

Kromosom

Gena

AA

Gena

Penyakit

Spectri

240

242

1q22q23

80 (Kb)

2429

SPTA1

HE,HPP,HS,NIHF

Spectri

220

242

14q22p24.2

>100

2137

SPTB

HE,HPP,HS,NIHF

2.1

Ankyrin

210

124

8p11.2

>120

1880

ANK1

HS

2.9

Adduci

103

30

4p16.3

737

ADDA

3*

AnExch1

90

1200

17q12p2.1

17

911

EPB3

HS,SAO,NIHF

4.1

P4.1

80

200

1p33p34.2

>250

588

EL1

HE

4.2

P4.2

72

250

15q15q21

20

691

ELB42

HS

PAS1

GlypA

36

-1 rb 4q31

>40

131

GYPA

PAS2

GlypC

32

-200 2q14421

14

128

GYPC

PAS3

GlypB

20

-200 4q31

>30

72

GYPB

4.9-8

HE

*protein integral; HPP: Hereditary pyropoikilocytosis; NIHF: Nonimmune hydrops fetalis; SAO:
Southeast Asian Ovalocytosis

Anemia
Anemia is a sign, not a separate disease
process.
Signs and symptoms may not be present
until the body is stressed.
Differentiate chronic anemia from acute
episode.

Treat signs and symptoms.

Maximize oxygenation and limit blood loss.
Establish IV therapy if indicated.

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Diseases of Erythrocytes

Brady; Paramedic Care Principles and Practice

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Diseases of Erythrocytes

Sickle Cell Disease

Normal red cells maintain
their shape as they pass
through the capillaries and
release oxygen to the
peripheral tissues (upper
panel). Hemoglobin polymers
form in the sickle cell cells
with oxygen release, causing
them to deform. The
deformed cells block the flow
of cells and interrupt the
delivery of oxygen to the
tissues (lower panel).
Kuliah Biomol 2010

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Diseases of Erythrocytes

Sickle Cell Disease (cont.)

Sickle cell crises
Vaso-occlusive
Musculoskeletal/abdominal pain
Priapism
Renal/cerebral infarctions

Hematological
Lowered hemoglobin
Splenic sequestration

Infectious

Management
Follow general treatment guidelines prn.
Consider analgesics.
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Diseases of Erythrocytes

Polycythemia
Overproduction of erythrocytes.
Occurs in patients > 50 years old or
with secondary dehydration.
Most deaths due to thrombosis

Results in bleeding abnormalities:

Epistaxis, spontaneous bruising, GI
bleeding.

Management:
Follow general treatment guidelines.
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THALASSEMIA

HEMOGLOBINOPATHY
Sandoz Atlas:
Cinical
Hematology,
Kuliah
Biomol
2010 1988 Gower Medical Publishing

43

Thalassemia
Sintesis hemoglobin

Eritrosit
Hb

300 juta/sel

Heme + globin
Fe + Porfirin

Rantai + non
(,,)

Thalassemia

H
e
m
e

2 2 = Hb A
>96%

2 2 = Hb A2
<3,5%

2 2 = Hb F
<2%

Molekul Hb
(tetramer)

Thalassemia

Pathophysiology:
(Thalassemia )

Defect sintesis of globin

excessive of globins
Deposited in cell
membrane
Deformability/liabilit
y
Fragilities

LIEN
Destruction
M, physic

HEMOLISIS

Thalassemia
Excessive of globin result in:

Denatura
tion

Excess globin
chain

Sp band3
crosslinkin
g

Band S
clusteri
ng

Degradat
ion

Abnorm
al
Spetcrin

associatio
n

Immune
removal

Abnorm
al Sp
low
binding

Deficiency
of Spect,
Ank
Abnormal
Sp 4.1
binding

Lipid
bilayer
damage
Sp
Actin,4.1
instability

Mechanichal
removal
Hypercoagulability
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Leukopenia/Neutropenia
Too few white blood cells or
neutrophils.
Follow general treatment guidelines
and provide supportive care.

Leukocytosis
An increase in the number of
circulating white blood cells, often
due to infection.
Leukemoid reaction
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Leukemia
Cancer of hematopoietic cells
Initial presentation

Acutely ill, fatigued, febrile and weak, anemic

Thrombocytopenia
Often have a secondary infection.

Management
Follow general treatment guidelines.
Utilize isolation techniques to limit risk of
infection.
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Lymphomas
Cancers of the lymphatic system
Hodgkin's
Non-Hodgkins

Presentation
Swelling of the lymph nodes
Fever, night sweats, anorexia, weight loss, fatigue,
and pruritis

Management
Follow general treatment guidelines.
Utilize isolation techniques to limit risk of
infection.
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Multifactorial (viral infection,

etc.....)
Complex-genes involvement
Chromosomal anomaly :
translocation
Diversity on clinical manifestation
Therapy approach gene therapy
???
Kuliah Biomol 2010

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Clotting Disorders

Thrombocytosis and Thrombocytopenia

Thrombocytosis
An abnormal increase in the number of platelets

Thrombocytopenia
An abnormal decrease in the number of
platelets
Sequestration
Destruction (ITP)
Decreased production

Management
Provide supportive care and follow general
treatment guidelines.

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Clotting Disorders

Hemophilia
Deficiency or absence of a blood clotting factor
Deficiency of factor VIII causes hemophilia A.
Deficiency of factor IX causes hemophilia B.
Deficiency is a sex-linked, inherited disorder.
Defective gene is carried on the X chromosome.

Signs & Symptoms

Numerous bruises, deep muscle bleeding, and
joint bleeding.

Kuliah Biomol 2010

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HEMOPHILIA

Kuliah
Biomol 2010
Victoria
princess
family, England

54

 Gene defect/protein FVIII (A), FIX (B)

Bleeding tendency (oozing)
Mild, moderated, severe
Herediter (X-linked) or spontan (approx. 30%)

Role of FVIII/IX in
hemostasis
Gene defect FVIII/IX
Deficiency FVIII/IX
Fibrin

Bleeding

Hemostasis mechanism
Hemophilia: defect protein of coagulation factor
VIII/IX
Vasokonstrk
si

Trombosi
t

Coagulation

close the hole

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Gene defect FVIII

1. Gene rearrangement

http://europium.mrc.rpms.ac.uk

40% hemophilia A severe

Cross over gene fragmented (meiosis)

2. Single-base substitutions
3. Sequence deletion
>100 kb = severe,
FVIII %, undetected Ag inhibitor?
<100 kb = mild/moderete

4. Sequence insertion

Management:
Causative: factor first!
(cryoprecipitate, concentrate, recombinant)
Bleeding correction (surgery)
Supportive (transfusion, etc)
Preventive:
Marriage counseling
Diagnosis of prenatal
Prophylaxis of bleeding
Complication:
Transfusion, inhibitor visibility, high
morbidity

Clotting Disorders

Hemophilia (cont.)
Management
Treat the patient similarly to others.
Administer supplemental oxygen.
Establish IV access.

Be alert for recurrent or prolonged bleeding, and

prevent additional trauma.

Von Willebrands Disease

Deficient component of factor VIII
Generally results in excessive bleeding.
Generally is not serious; provide supportive
care.
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Disseminated Intravascular
Coagulation
System activation of coagulation cascade.
Results from sepsis, hypotension, OB
complications, severe tissue or brain
injury, cancer, and major hemolytic
reactions.

Multiple Myeloma
Cancerous disorder of plasma cells.
Pathologic fractures are common.
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