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Clinical Pathology > Week 1 - Erythropoiesis and the erythrogram > Flashcards

Week 1 - Erythropoiesis and the erythrogram Flashcards

1
Q

Define the term erythron

A

Definition: the red blood cells and their precursors in the bone marrow.

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2
Q

All erythroid cells in an animal include?

A

All erythroid cells (red blood cells or RBCs) in an animal
* Includes precursor cells, and RBCs in blood vessels and sinuses in the spleen, liver, bone marrow

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3
Q

What % of an erythrocyte is made up of hemoglobin?

A

95%

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4
Q

Describe the structure of a hemoglobin molecule.

A
  • Each hemoglobin (Hgb) molecule is a tetramer
  • 4 globin chains:
  • 2 α-chains & 2 β-chains
  • Each globin chain is linked to a
    separate heme molecule that
    binds O2
  • Iron in the ferrous state (Fe +2 ) is
    incorporated into each heme
    molecule
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5
Q

What is the function of Hgb?

A
  • To transport oxygen (O2) from lungs to tissues
  • In health, Hgb is 100% saturated with O2 in arterial blood
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6
Q

How is iron able to transport O2?

A
  • Iron must be in the reduced state (Fe2+ ) to transport O2
  • Hemoglobin bound to Fe 3+ cannot carry oxygen

Hgb + Fe 2+ only way to transport O

Some toxins prevent iron from being reduced, so iron will not be transported. Need iron!!

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7
Q

Why is iron important?

A
  • Because hemoglobin synthesis depends on IRON!
  • Series of enzymatic reactions for Hgb synthesis
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8
Q

How is iron distributed throughout the body?

A
  • Major sites
    1. Erythrocyte Hgb (50-70%)
    2. Tissue storage (25-40%) -as ferritin-
    3. Remainder in other molecules (myoglobin, cytochromes, enzymes)
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9
Q

How is hemoglobin synthesized?

A

so won’t have completely formed heme molecule

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10
Q

Describe the condition called Porphyrias. What is it caused by? What does it lead to? What species are affected?

A
  • Rare, hereditary disease in cattle, pigs, cats, & humans
  • Caused by deficiency in hemoglobin synthesis enzyme URO-synthase
  • Leads to porphyrin accumulation with incomplete heme synthesis
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11
Q

What condition is pictured below?
How would you describe what can be seen in this image?

A

Porphyria
Marked pink discoloration of the teeth

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12
Q

What condition is pictured below? How would you describe what can be seen in this image?

A

Porphyria
Bight pink-red fluorescent
color of bone marrow and cartilages viewed under ultraviolet (UV) light.

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13
Q

What are the four major erythrocyte metabolism pathways?

A
  1. Embden-Meyerhoff pathway (glycolysis)
  2. 2,3-DPG (AKA: Rapoport-Luebering) pathway
  3. Pentose phosphate pathway
  4. Methemoglobin reductase pathway
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14
Q

Describe the Embden-Meyerhoff pathway (glycolysis)
1. What does it generate?
2. What are the 2 relevant enzymes in this process?

A
  1. Generates ATP to maintain membrane function and
    integrity
  2. Generates NADH to reduce methemoglobin
    (Fe +3+Hb –> Fe +2+Hb)
  3. Contains 2 clinically relevant enzymes:
    - Pyruvate kinase (PK)
    - Phosphofructokinase (PFK)
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15
Q

What is the function of the 2,3-DPG (AKA: Rapoport-Luebering) pathway?

A

Aids in O2 delivery to tissues when anemia (reduced RBC mass) is present

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16
Q

What does the Pentose phosphate pathway generate?

A

Generates NADPH (cofactor for glutathione reductase)

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17
Q

What is the main function of the Methemoglobin reductase pathway?

A

Reduces iron (Fe+3 –> Fe +2 )

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18
Q

What is the main purpose of the Pentose phosphate pathway and the methemoglobin reductase pathway?

A

These 2 pathways provide protection against oxidative stress:
* Toxic exposures can overcome these pathways (more on this later in the semester)

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19
Q

Describe the condition Phosphofructokinase
(PFK) deficiency. What species are affected?

A
  • English Springer Spaniels, American Cocker Spaniels
  • Shortened RBC lifespan due to impaired ATP production
  • Decreased 2,3-DPG concentration
  • Alkalemia-induced hemolytic anemia (hyperventilation when
    stressed/excited)
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20
Q

Describe the condition called Pyruvate kinase (PK) deficiency. What species are affected?

A
  • Basenjis, Beagles, Chihuahuas, Dachshunds; Abyssinians, Somalis
  • Shortened RBC lifespan
  • Accumulation of 2,3-DPG and reduced ATP production
  • Bone marrow failure
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21
Q
  • Erythropoiesis = ________ production
  • Occurs mostly in the ______ _______. ___________ enzyme facilitates this process. Main source of this enzyme is the _______, sometimes the _____.
  • Blast-forming unit-erythroid (BFU-E) –> committed stem cells (CFU-E)
  • CFU-E –> rubriblasts (?)
  • Stimulated by erythropoietin (Epo) and other cytokines
  • Epo is produced in _____ kidney and _____ liver
A

erythrocyte, bone marrow, Erythropoietin, kidney, liver

first microscopically recognizable erythroid cell, adult, fetal

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22
Q

What is the name of the only RBC you should see in circulation?

A

Should only see erythrocyte in circulation.

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23
Q

List and describe the erythrocyte development stages.

A
  • Rubriblast
  • Prorubricyte
  • Rubricyte: basophilic and polychromatophilic
  • Last mitotic stage
  • Metarubricyte
  • Last stage in which the developing RBC is nucleated
  • Reticulocyte
  • Anucleated
  • Larger and more basophilic (more blue) than a mature
    erythrocyte – still has stainable cytoplasmic RNA
  • Mature erythrocyte
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24
Q
  • Rubriblast –> erythrocyte
    § Cells produce _____ for hemoglobin synthesis
    § Cells undergo ______ to produce more & smaller cells that have progressively more _________
    § As hemoglobin synthesis increases in the developing cell, DNA synthesis ______ and ______ mitotic divisions occur
    § Takes ~ ___ days for erythropoietic progenitor cell –> reticulocyte
    § Reticulocyte is released from _____ ________ into circulation and circulates for __-__ days before becoming a fully mature erythrocyte
A

mRNA, mitoses, hemoglobin, decreases, fewer, 5, bone marrow, 1-2

25
Q

What factors control the process of erythropoiesis?

A
  • Nutritional factors (most important one)
  • Hormonal factors
  • Bone Marrow factors
26
Q

Describe the nutritional factors that control erythropoiesis, specifically nucleic acid metabolism and cytoplasmic maturation and hemoglobin formation.

A
  1. Nucleic acid metabolism
    * Vitamin B12 (Aka Cobalamin)
    * Folate
  2. Cytoplasmic maturation and hemoglobin formation
    * Amino acids
    * Vitamin B6
    * Iron (critical for Hgb synthesis)
    * Copper
27
Q

Describe the hormonal factors that control erythropoiesis.

A
  • Hormonal factors
  • Erythropoietin (Epo) produced in kidney is the main signal for erythropoiesis to occur in the bone marrow.
    Ery can also occur in spleen for precursor cells to become mature cells (to replace dead RBC).
28
Q

Describe the bone marrow factors that control erythropoiesis.

A

Bone Marrow factors
* Stem cells
* Stroma (if you don’t have enough room for this process to occur).
* Blood supply

29
Q

What is the lifespan of an erythrocyte in the following species? Why is knowing this imoportant?
1. birds
2. cats
3. dogs
4. horses
5. cows

A

RBC lifespan in circulation: ranges
* 35 days (birds),
* 70 days (cats),
* 110 days (dogs)
* 145 days (horses)
* 160 days (cows)
§ Importance: may not see anemia right away

30
Q

How are erythrocytes destroyed?

A
  • Two mechanisms of RBC removal:
    § Major: phagocytosis by macrophages,
    mainly in the spleen
    § Minor: intravascular lysis with release of
    hemoglobin into plasma
31
Q

How is Hgb degraded by macrophages?

A

Excess amount of byproducts –> icterus

32
Q

What is the main function of Erythron?

A
  • Erythron main function: transport O2 which is
    mediated by Hgb
  • Hbg is transported by erythrocytes
  • Hgb consists of heme & globin
33
Q

Define the term erythrogram.
What does a CBC assess?

A
  • Component of a complete blood count (CBC)
  • CBC Assesses erythrocytes, leukocytes, and platelets
  • Blood is collected into a purple-top tube containing
    EDTA and processed through a hematology analyzer
  • EDTA: anticoagulant
  • Chelates calcium to stop the clotting cascade
34
Q
A
35
Q

RBC count is reported in? Measured by?

A
  • RBC (Red blood cell count)
    § Reported in millions
    § Measured by the instrument
36
Q

Hgb is reported in? Measured by?

A
  • Hgb (Hemoglobin)
    § Typically reported in g/dL
    § Grams of Hgb per 100 mL of blood
    § Measured via spectrophotometry methods
37
Q

HCT is defined as?
How do you obtain the calculated value?
If HCT is below the RI? Above?

A
  • HCT (Hematocrit)
    § Percentage of blood volume filled by erythrocytes
    § Calculated value (MCV x RBC) / 10
    § If HCT is below the RI, the patient is anemic
    § Usually will have concurrent decreased Hgb and RBC
    § If HCT is above the RI, the patient has an erythrocytosis
38
Q
  • PCV = packed cell volume
  • Centrifugal method
  • Very accurate with a small error margin
  • More accurate than _____ if any RBC parameters are falsely altered (e.g. MCV, RBC count)
  • Rule of thumb: HCT and PCV should be ~__x Hgb concentration
  • HCT and PCV should differ by maximum of 3%
A

HCT, 3

39
Q
A

Erythrogram terminology:
Wintrobe’s indices
* MCV: Mean corpuscular/cell volume
* MCH: Mean corpuscular hemoglobin
* MCHC: Mean corpuscular hemoglobin concentration

40
Q

MCV is defined as?
How is this value expressed?
How is this value calculated?
If the MCV is below the RI? Above?

A

MCV: mean corpuscular/cell volume
* Volume per average erythrocyte
* Expressed in fL
* Calculated by some instruments & measured by
others
* (Hct x 10) / RBC
* If MCV is below the RI, there is a microcytosis
* If MCV is above the RI, there is a macrocytosis

41
Q

MCH is defined as?
How is this value expressed?
How is this value calculated?
If the MCH is below the RI? Above?

A

MCH: mean corpuscular hemoglobin
* Quantity of Hgb per average erythrocyte, expressed
in picograms (pg)
* Calculated value
* (Hgbx10)/RBC
* If MCH is below the RI, there is hypochromasia
* If MCH is above the RI, there is hyperchromasia

42
Q

Inaccuracies in RBC count will lead to?

A

Inaccuracies in RBC count
will lead to
MCV and MCH inaccuracies

43
Q

MCHC is defined as?
How is this value expressed?
How is this value calculated?
If the MCHC is below the RI? Above?

A

MCHC: mean corpuscular hemoglobin concentration
* Cellular Hgb concentration per average erythrocyte
* Expressed as grams of Hgb per 100 mL erythrocytes
(g/dL)
* Calculated value
* (Hgbx100)/Hct
* More accurate than MCH
* If MCHC is below the RI, there is hypochromasia
* If MCHC is above the RI, there is hyperchromasia

44
Q

What can influence MCV and MCHC values?

A

Pre-analytical factors can influence MCV and MCHC values
1. Short sample (underfilled EDTA tube) or low plasma osmolality (low Na and Cl)
- use correct size tube for blood/sample so that machine can run correctly
- Falsely decreased MCV and increased MCHC
2. Increased plasma osmolality (high Na and Cl)
- Falsely increased MCV and decreased MCHC

45
Q

What does it mean when a patient has a low number of Reticulocytes? High number? What is the exception to this rule?

A
  • In health, low numbers normally present in
    circulation in most species
  • When increased, indicates increased RBC
    production (regenerative anemia)
  • Typically interpreted in context of an anemia
    (regenerative or non- regenerative)
  • *Exception: horses release few (if any) reticulocytes
  • Cattle and small ruminants: basophilic stippling

Make sure machines are calibrated to the proper species due to differences in cell morphology, etc.

46
Q

How do you count reticulocytes?

A
  • Instrument uses a special dye for reticulocyte RNA and measures what percentage of all erythrocytes are reticulocytes
  • Can also manually count on a blood smear stained with new methylene blue
47
Q

How do you obtain the absolute reticulocyte count (absRC).

A
  • absRC = reticulocyte percentage x RBC count (in millions)
  • Reported by some hematology analyzers
  • Can be calculated
48
Q

How do you obtain the corrected reticulocyte percentage (CRP)?

A
  • CRP = reticulocyte% x (patient’s HCT/normal HCT for that species)
  • Normal HCT used for
    § canines = 45%
    § Felines = 35%
49
Q
  1. How are nucleated RBC’s (nRBCs) typically expressed?
  2. The number of nRBC’s can be increased in what situations?
  3. nRBC’s are usually?
A

Do not see them often. May see a couple or a few.
* Typically expressed as number per 100 WBCs
counted
* See an increased number in these situations:
* strongly regenerative anemias or
* damage to bone marrow endothelium
* Usually metarubricytes or rubricytes, but can
include earlier stages in very strongly regenerative
anemias or leukemia

50
Q

What are the arrows pointing to in this image?

A

Reticulocytes

51
Q

What is the arrow pointing to in this image?

A

nRBCs

52
Q
  1. What does the red cell distribution width (RDW) represent?
  2. How do you obtain the calculated value?
  3. What does RDW indicate?
  4. Is the RDW indicated by all analyzers?
A
  • Represents coefficient of variation of RBC volume
    distribution
  • Calculated value: RDW = (SDMCV/MCV)*100
  • Indicates degree of anisocytosis (variation in RBC
    size)
  • Not reported by all analyzers
53
Q
  1. What does plasma protein represent?
  2. How is plasma protein determined?
  3. What is it interpreted with?
  4. If the plasma protein is above the RI? Below?
A
  • Plasma protein (aka TP or “total solids”)
  • Reported as g/dL
  • Determined by refractometry
  • Usually interpreted in conjunction with RBC parameters
  • If below the RI, indicates Hypoproteinemia
  • By loss or lack of production
  • If above the RI, indicates Hyperproteinemia
  • By increased production or hemoconcentration
54
Q
  1. How is fibrinogen measured?
  2. Is this routinely reported?
  3. If fibrinogen is below the RI? Above?
A
  1. Measured in large animals by heat precipitation method.
    • Not routinely reported on a CBC in small animals
      * If below the RI, it is called Hypofibrinogenemia
      * indicates loss or lack of production
      * If above the RI, it is called Hyperfibrinogenemia
      § Indicates increased production (inflammation)
      § Considered Positive acute phase protein
      § indicates hemoconcentration
55
Q

Explain the two ways in which you can calculate the absRC.

A
56
Q
A

high MCV = macrocytic
MCH high and MCHC is high = macrochromic BUT even though MCH is flagged, the MCHC is more accurate. Th

Macrocyctic normochromic anemia?

9 x 1.2 = 10.8

57
Q
A

low MCV = microcytic

other params norm = normochromic

58
Q
A

norm MCV = normocytic

MCH low = microchromic

normocytic microchromic

Clinical Pathology (31 decks)

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