Hemetology bchm:

In order for a child to have sickle cell disease both parent must be carriers. The carrier status of prospective
parents can be established by hemoglobin electrophoresis.
Glucose 6-phosphate dehydrogenase deficiency is a defect in the HMP shunt that impairs glutathione
reduction due to failure to produce NADPH. Glutathione reductase deficiency causes a similar clinical
picture and is pathophysiologically similar to G6PD deficiency.
HbF dominates in newborns. It consists of two alpha and two gamma protein subunits (2Y2), has a high
affinity for oxygen and is produced during the final seven months of gestation. Switching to HbA (22)
occurs during the first six months of life.
Pyruvate kinase deficiency causes hemolytic anemia due to failure of glycolysis and resultant failure to
generate sufficient ATP to maintain erythrocyte structure. In this case, splenic hypertrophy results from
increased work of the splenic parenchyma, which must remove these deformed erythrocytes from the
circulation.
Transketolase and transaldolase carry out the nonoxidative reactions of HMP shunt. Some cells do not use
the oxidative phase reactions to produce cytosolic NADPH, but all cells can synthesize ribose from
fructose-6-phosphate using the nonoxidative reactions.
Normally, 2, 3-DPG forms ionic bonds with the two beta subunits of HbA in the tissues after hemoglobin
has been deoxygenated. Fetal hemoglobin binds oxygen with a higher affinity due to its inability to interact
with 2, 3- DPG. Ultimately, the fetal hemoglobin must be able to extract 02 from maternal hemoglobin in
the placenta.
Increased 2,3-BPG concentrations within erythrocytes enable increased oxygen delivery in the peripheral
tissues in the presence of lower blood oxygen concentration because 2,3-BPG decreases the affinity of
hemoglobin for oxygen. 2,3-BPG is produced from 1,3-BPG by the enzyme phosphoglycerate mutase. This
reaction consumes energy that would have been otherwise used by the erythrocyte to produce energy in the
form of ATP.
Methemoglobinemia causes dusky discoloration to the skin (similar to cyanosis), and because
methemoglobin is unable to carry oxygen, a state of functional anemia is induced. The blood partial
pressure of O2 however, will be unchanged in this condition because oxygens partial pressure is a
measure of O2 dissolved in the plasma and is not related to hemoglobin function.
Heme oxygenase converts heme to biliverdin, a pigment that causes the greenish color to develop in bruises
several days after an injury.
Vitamin K assistance of glutamine residue carboxylation is essential for some clotting factor production.
HbS contains valine in place of glutamic acid in the 6th amino acid position of the beta subunit. This
promotes hydrophobic interaction among hemoglobin molecules and results in polymerization of HbS
molecules and red blood cell distortion.
Folate deficiency inhibits the formation of deoxythymidine monophosphate (dTMP) which limits DNA
synthesis and promotes megaloblastosis and erythroid precursor cell apoptosis. Because thymidine
supplementation can moderately increase dTMP levels, it can reduce erythroid precursor cell apoptosis.
While DNA synthesis occurs in the 5' to 3' direction on both strands the leading and lagging strands are
constructedin both the 5' to 3' and 3' to 5' directions respectively. The lagging strand is synthesized
discontinuously and is composed of short stretches of RNA primer plus newly synthesized DNA segments
called Okazaki fragments. Thus the lagging strand requires the repetitive action of DNA primase and DNA
ligase.
Nitrites are oxidizing agents that are effective in the treatment of cyanide poisoning due to their ability to
cause methemoglobinemia. Methemoglobin contains ferric rather than ferrous iron. Cyanide binds to ferric
iron more avidly than to mitochondrial cytochrome enzymes, which saves these mitochondrial enzymes
from cyanides toxic effect.
CO binds to hemoglobin with an affinity that is 220 times that of oxygen for hemoglobin. The binding of
CO and O2 to hemoglobin are reversible. CO, therefore, competes with O2 for binding on the heme iron of
hemoglobin.
The Kozak sequence plays a role in the initiation of translation. A mutation three bases upstream from the
start codon (AUG) in this sequence is associated with thalassemia intermedia whch results in
hyperchromic, microcytic anemia.

The thalassemias result from mutations that cause defective mRNA processing which leads to deficiency of
certain protein chains required for hemoglobin synthesis. Beta thalassemia minor is typically an
asymptomatic disorder. Laboratory tests will show a mild hypochromic microcytic anemia with increased
HbF, HbA2, and target cells.
A left shift of the hemoglobin oxygen dissociation curve indicates increased hemoglobin O2 affinity and
can be caused by increased pH, decreased 2, 3-DPGI and decreased temperature. A left-shift of the oxygendissociation curbe means that is relatively less available to tissues.
With the exception of vitamin B12, the bodys stores of most water soluble vitamins are rapidly depleted
without dietary intake. In contrast hepatic stores of vitamin B12 may last up to several years. Liver stores
of vitamin A last about six months without dietary intake. Eat stores of vitamin D can last a few months in
the absence of dietary intake or sunlight exposure. Severe vitamin K deficiency rarely results from poor
dietary intake because colonic bacteria produce functional forms of vitamin K.
The symptoms of difficulty in swallowing (dysphagia) and disfigured fingernails (spoon nails or
koilonychia) are specific for iron deficiency anemia.
Cobalamin (Vitamin B12 deficiency results in homocystinemia due to impaired methionine synthesis.
Homocystinuria occurs in cobalamin (Vitamin B12 deficiency because homocysteine methyltransferase,
the enzyme that converts homocysteine and methyltetrahydrofolate to methionine and tetrahydrofolate,
requires B12 as a cofactor.
UGA, UAG, and UAA are stop codons, and mutations producing abnormally placed stop codons are called
nonsense mutations.
HbC is caused by a missense mutation that causes a substitution of glutamine with lysine in the beta globin
chain, resulting in increased positive charge of the molecule. Thus, HbC moves more slowly than both HbA
and HbS on hemoglobin gel electrophoresis.
Glucose 6-phosphate dehydrogenase deficiency is a common X-linked disorder of the hexose
monophosphate pathway that results in episodes of hemocytic anemia due to oxidative stress.
Hemoglobin S (HbS) aggregates in the deoxygenated state. HbS polymers form fibrous strands that reduce
red blood cell membrane flexibility and promote sickling. Sickling occurs under all conditions associated
with anoxia including low pH and high 2I3-DPG. These inflexible erythrocytes predispose to
microvascular occlusion and micro infarction.
Decreased heme concentration results in an increase in hepatic ALA synthase activity, which in turn, leads
to increased formation of 5-aminolevulinic acid and porphobilinogen. Increased formation of 6aminolevulinic acid and porphobilinogen occurs because heme normally serves to inhibit the synthesis of
ALA synthase.
Western blotting is used to identify proteins, Northern blotting identifies specific RNA sequences and
Southern blotting identifies specific DNA sequences in an unknown sample.
The P50 refers to the partial pressure of oxygen where hemoglobin is 50% saturated. A decrease in the P50
means that hemoglobin has an increased oxygen affinity. An increased oxygen-affinity of hemoglobin
causes less oxygen to be released in the tissues, and results in hypoxia, then reflex polycythemia.
Exertional dyspnea, pneumonia resulting in life threatening acute chest syndrome, and recurrent abdominal
and bone pain are clinical features of sickle cell anemia. Sickle cell anemia results from a point mutation
that causes valine to substitute for glutamic acid in the sixth position of the b-globin chain of hemoglobin
Maturing erythrocytes lose their ability to synthesize heme when they lose their mitochondria.
Mitochondria are necessary for the first and final three steps of heme synthesis.
The liver takes up indirect (unconjugated) bilirubin through a passive process and secretes direct
(conjugated) bilirubin through an active process. Unconjugated bilirubin is virtually insoluble in water at
physiologic pH and is tightly complexed to serum albumin while in the circulation. This form cannot be
excreted in the urine even when blood levels are high.
HbF contains y-globin instead of -globin. Patients with homozygotic 13-thalassemia (-thalassemia
major) are asymptomatic at birth due to the presence of y-globins and HbF Switching to HbA production
and the cessation of Y-globin synthesis precipitates the symptoms of -thalassemia.
In the lungs, hemoglobin binds oxygen and releases protons; while, in the tissues, it releases O2, and
acquires protons. Deoxyhemoglobin is stabilized by ionic bonding of 2, 3 DPG to the two beta subunits and
by salt bridges between N-terminal histidine residues in each globulin.
The individual subunits of the hemoglobin molecule are structurally analogous to myoglobin; if separated
the subunits will demonstrate a hyperbolic oxygen-dissociation curve similar to that of myoglobin.