Maria Isabel Arango Sánchez-Tercer semestre

1. M A R I A I S A B E L A R A N G O S Á N C H E Z
E S T U D I A N T E D E M E D I C I N A
T E R C E R S E M E S T R E - 2 0 1 9

2. THALASSEMIA
Thalassemia is an inherited blood disorder characterized
by less hemoglobin and red blood cells resulting from the
synthesis of abnormal globin chains.
Classification:
According to which globin chains it is affected: Can be α-
thalassemia or β-thalassemia, the last one it’s the most common
form of this disease
According to the intensity of the symptoms it can be
major, intermedia and minor
Patients may present with any of the following symptoms typical of anemia and/or
extramedullary erythropoiesis like: Fatigue -Asthenia -Anorexia -Exercise
intolerance -Recurrent fevers -Feeding problems -Dyspnea on exertion or at rest -
Abdominal pain -Dark urine -Bone pain or tenderness .
The ethnic groups with the highest incidence are
Asians and Africans

3. HEMOGLOBIN E HEMOGLOBIN RUSH
Anomalous hemoglobin, it is due to a substitution of
glutamic acid for lysine in codon 26 of the β-globin
gene.
HbE determines a reduction in the synthesis of the
β-E chain and that is why the phenotype it’s an
intermedia β-thalassemia.
Heterozygous composed of HbE and a β-thalassemia
gene is the most frequent severe variant of β-
thalassemia
It is an anomalous hemoglobin ,in this one
the glutamic acid is replaced by glutamine in
the β-chain, β101 (G3) .
The Hb Rush mutation causes the central
cavity of the protein that is usually neutral in
charge to become more positive, causing cell
lysis
The Hb Rush has been described to be
responsible for mild hemolytic anemia.

4. OVERALL OBJECTIVE
•“We provide the first evidence of
Hb Rush in association with HbE
and B-thalassemia mutations
leading to thalassemia intermedia
phenotypes”.
Tomado de :https://www.tandfonline.com/action/journalInformation?journalCode=yhem20

5. MATERIALES Y
MÉTODOS
The probands were eleven patients
from 8 unrelated ethnic families in
Yunnan province,Southwestern China
Anticuagulated periferal blood
measured whit an automated
hematology analyzer.
Hemoglobin fractions separations
whith agarosa gel in an alkaline
medium.
Imágenes tomadas de :https://mx.depositphotos.com/87102444/stock-photo-molecular-structure-of-hemoglobin.htm-http://atlas.gechem.org/es/component/k2/item/648-centrocito-en-sangre-periferica
https://www.flaticon.es/icono-gratis/figura-basica_62122

6. ARMS-PCR
Basis:is Use a sequence-specific PCR primers that
allow amplification of test DNA only when the target
allele is contained within the sample. Following an
ARMS reaction the presence or absence of a PCR
product tell us if is present or absence of the target
allele.
Utility: Detecting any mutation
involving single base changes
or small deletions.

7. PCR-RFLP
Utility: They are useful as location markers in
genomic DNA that allows me to identify the
composition of the DNA and if it has undergone
any changes
Basis: Restriction enzymes are used to cut DNA
molecules in known places causing RFLPs that are
used as markers in genetic maps

8. SEQUENCING Electrophoresis
Bases: It is an enzymatic method that allows
to determine the sequence of the mold as its
complementary strand is synthesized.
Utility:It helps us to determine the
nucleotide sequence of a DNA fragment.
Utility: Separate molecules
Bases:
Under the influence of an electric field in a
liquid medium the charged particles are placed,
which will begin to migrate
Imagen tomada de : http://biomodel.uah.es/tecnicas/elfo/inicio.htm

9. RESULTS
•
In image A we see that in lanes 1,2 and 3
there is a mutation which produces 181
bp fragment, the apparition of this
fragment causes HB Rush
In image B we see the DNA sequence
that shows a heterocygous type
mutation that gives me an affirmative
result for the presence of Hb Rush
Imágenes tomadas de :https://www.tandfonline.com/loi/yhem20

10. Electrophoresis shows
hemoglobin separation
patterns and we can observe
that’s Rush/E and Rush/CD 71
they advanced more than the
others ,showed that the
electrophoretic medium and
pH conditions interfere with
the separation profile of Hb
Rush.
Imágenes tomadas de :https://www.tandfonline.com/loi/yhem20

11. Here we can see that when there is Hb Rush in the company of HbE, the amount of Hb Rush that is
produced is further increased and the appearance of other anomalous hemoglobin such as Hb F is
stimulated, which further decreases the amount of normal B chain production
Imágenes tomadas de :https://www.tandfonline.com/loi/yhem20

12. The Hb Rush simple heterozygote presented hemolytic animea whir hemoglobin concentrations of
85-109g/L .The compound heterozygosity exhibited microcytic hipochromic anemia whit HBG,MVC
and MCH values much lower and alarming (P7-P8-P1-P2)
Imágenes tomadas de :https://www.tandfonline.com/loi/yhem20

13. DISCUSSION
AUTHOR CONCEPT YES OR NOT
Thein SL. Genetic basis and genetic modifiers
of beta- thalassemia and sickle cell disease.
Adv Exp Med Biol. 2017;1013:27–57.
Our study provides new evi- dence and an
example model of environmental influence
on the clinical expression of thalassemia
intermedia. Hb F induction was also
observed in the Hb Rush compound
heterozygotes, thus suggesting another
possible modifying factor in the thalassemia
intermedia
Silva MR, Sendin SM,Velloso-Rodrigues C, et
al. Unstable hemoglobin Rush [beta 101(G3)
Glu>Gln, HBB: c.304G>C] in a Brazilian
family with moderate hemolytic anemia.Ann
Hematol. 2012;91(7):1091–1096.
An Hpy188I restriction enzyme site is
involved in the nucleotide substitution of Hb
Rush, and a PCR-RFLP protocol may be used
to detect the variant
Williamson D.The unstable
haemoglobins. Blood Rev.
1993;7(3):146–163.
Although Heinz bodies are charac-
teristic of unstable hemoglobins and
hemolytic anemia .

14. CONCLUSION
The coexistence between Hb Rush and anomalous hemoglobins such as
HB E causes a change in the behavior of Hb Rush, significantly increasing
its percentage present in erythrocytes, which results in the appearance of
major and intermediate thalassemia.
Molecular biology is a fundamental area of medical knowledge because it
provides us with the necessary tools to understand the processes that
occur in the human body from the functioning and interaction between
the molecules and cells that compose it, which allows us to understand
exactly where the damage is found in a pathology and therefore develop
better mechanisms to fight them.