Unit 2 - Erythropoiesis Flashcards

1
Q

Erythropoiesis Sequence

A

HSC –> MPP –> CMP –> CFU-GEMM –> BFU-E

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2
Q

Where is EPO produced in adults

A

Kidneys in the peri tubular interstitial cell

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3
Q

What happens to EPO in kidney disease?

A

Levels decrease

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4
Q

Where is EPO produced in embryos?

A

Liver

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5
Q

Where is the backup production for EPO in adults? How much is produced?

A

Liver
Not much

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6
Q

What is the first cell recognized as a young erythrocyte by light microscopy?

A

Pronormoblast
Rubiblast

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7
Q

How long is the maturation time from blast to erythrocyte?

A

5-7 days

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8
Q

What are nucleated RBC precursors called in the bone marrow, collectively?

A

Erythroblast

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9
Q

Cytoplasm trend in RBC as it matures

A

Deep blue to red

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10
Q

How does the nucleus size of an RBC move as it matures?

A

Large to small to gone

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11
Q

Chromatin trend in RBC as it matures

A

fine to coarse

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12
Q

Cell stages (normoblast version)

A

Pronormoblast
Basophilic normoblast
Polychromatophilic normoblast
Orthochromic normoblast
Reticulocyte/Polychromatophilic RBC
Mature Erythrocyte

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13
Q

Cell stages (other version)

A

Rubriblast
Prorubricyte
Rubricyte
Metarubricyte
Diffusely basophilic RBC
Discocyte

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14
Q

Pronormoblast/Rubriblast size

A

20 micrometers

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15
Q

Color of pronormoblast/rubriblast cytoplasm

A

Very blue/purplish
Darker at edge

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16
Q

Appearance of nucleus in Pronormoblast/Rubriblast

A

Large nucleus
Fine parachromatin/euchromatin

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17
Q

Basophillic Normoblast/Prorubricyte size

A

16-18 micrometers

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18
Q

Extra features inside Pronormoblast/Rubriblast

A

Prominent nucleoli
Unstained golgi body

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19
Q

Appearance of nucleus in Basophilic Normoblast/Prorubricyte

A

Some coarseness

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20
Q

Appearance of cytoplasm in Basophilic Normoblast/Prorubricyte

A

Still very blue, but hgb starts to give a pinkish tinge

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21
Q

Extra features of a Basophilic Normoblast/Prorubricyte

A

Nucleoli less visible
Almost can’t see Golgi body

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22
Q

Polychromatophilic normoblast/Rubricyte size

A

15 micrometers

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23
Q

Appearance of nucleus in polychromatophilic normoblast/Rubricyte

A

Clumped areas of chromatin
Some patches of parachromatin

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24
Q

Appearance of cytoplasm in polychromatophilic normoblast/rubricyte?

A

Blue RNA
Pink Hb
Dirty gray-ish pink

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25
Q

What is the last stage of an erythroblast that has a nucleoli and Golgi body?

A

Basophilic Normoblast
Prorubricyte

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26
Q

Orthochromic Normoblast/Metarubricyte size

A

12 micrometers

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27
Q

Appearance of Orthochromic Normoblast/Metarubricyte nucleus

A

More clumped
PYKNOTIC!

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28
Q

What is the name of the appearance of the nucleus of an orhochromic normoblast/metarubricyte?

A

Pyknotic

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29
Q

Appearance of orthochromic normoblast/metarubricyte cytoplasm

A

Mostly pink
Slight blue tinge
mRNA still synthesizing Hgb

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30
Q

What can’t an orthochromic normoblast/metarubricyte do?

A

Mitose

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31
Q

Which is the first erythroblast that can’t mitose?

A

Orthochromic Normoblast
Metarubricyte

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32
Q

Which is the last erythroblast that can mitose?

A

Polychromatophilic Normoblast
Rubricyte

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33
Q

What does an orthochromic normoblast/metarubricyte do with its nucleus?

A

Pushes it to the side as it gets ready to push it out

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34
Q

Reticulocyte size

A

9-11 micrometers

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35
Q

Appearance of cytoplasm of a reticulocyte/Diffusely basophilic RBC?

A

Blue tinge
mRNA still synthesizing Hgb

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36
Q

Appearance of nucleus of a reticulocyte/diffusely basophilic RBC?

A

None

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37
Q

How long do reticulocytes remain in the bone marrow?

A

1-2 days

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38
Q

How long does it take for reticulocytes to mature in the peripheral blood?

A

~1 day

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39
Q

mRNA still present gives off what color? Retics

A

Purple

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40
Q

What is a reticulocyte called on a blood smear?

A

Polychromatophilic

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41
Q

How does the RNA stain in a relic?

A

The ACIDIC RNA stains BLUE
Basic (blue) component of stain loves the acidic (basophilic) part of the cell

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42
Q

Can supravital stains penetrate dead cells?

A

No

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43
Q

What is the supra vital stain used to observe the RNA in retics?

A

New Methylene Blue

44
Q

What does new methylene blue do to the reticulocyte appearance?

A

mRNA in endoplasmic reticulum looks like coarse granules

45
Q

How many inclusions are required to call a reticulocyte?

A

At least TWO

46
Q

Mature erythrocyte size

A

7-9 micrometers

47
Q

What removes old RBCs?

A

Reticuloendothelial/phagocytic system

48
Q

Where are old RBCs removed?

A

Spleen
Liver
Marrow

49
Q

What RBCs are found in adult peripheral blood normally?

A

Retics
RBCs

50
Q

What RBCs are found in newborn peripheral blood normally?

A

Polychromatophilic normoblast/Rubricyte
Orthochromic normoblast/Metarubricyte
Reticulocytes
Erythrocytes

51
Q

What could be possible if nucleated RBCs are in periphera blood

A

Patient is a newborn

Patient has a disease or anemia

52
Q

What tests are ordered to evaluate Erythropoiesis?

A

CBC
Peripheral blood smear
Reticulocyte count

53
Q

What does CBC tell you?

A

WBC count
RBC count
PLT count
Hct
Hgb
Indices (MCV/MCH/MCHC)

54
Q

What is one of the most valuable lab tests for evaluating erythropoiesis?

A

Peripheral blood smear

55
Q

Normal portion of RBCs that are retics?

A

1%
0.5%-2.0%

56
Q

When would retics be elevated?

A

Blood Loss
RBC destruction (hemolysis)

57
Q

When would retics be decreased?

A

Bone Marrow Production issue
Transfusion

58
Q

Why does transfusion cause a decrease in retics?

A

RBCs in blood bag are still alive and continue to mature

59
Q

How long is blood for retics stable at RT in EDTA?

A

6 hours

60
Q

How many RBCs are counted to get retic count?

A

1000

61
Q

Retic supra vital stains

A

New Methylene Blue
Brilliant Cresyl Blue

62
Q

How is the classic manual reticulocyte counting done?

A

Mix with supra vital stain
Incubate to allow staining
Count number of retics/1000 RBCs for %

63
Q

What is a miller disc?

A

A device in the ocular that places squares over the field

64
Q

How big is the small square of a miller disc?

A

1/10 the size of the large square

65
Q

Miller Disc Retic count procedure

A

B square - count total red cells (mature & retic)
A square - count only retics (+B square)

Count 111 RBCs in B square
Divide # of retics counted by 10 for retic %
(#retics/10) = %

66
Q

How do you know what cells to count and not count on the miller disc procedure?

A

Count a cell if it falls on LEFT or UPPER LINE
Don’t count a cell if it falls on RIGHT or BOTTOM LINE

67
Q

What cells do you NOT count on a miller disc?

A

BOTTOM and RIGHT line

68
Q

What cells DO you count on a miller disc?

A

LEFT and UPPER line

69
Q

Sources of error in reticulocyte evaluation

A

Poor mixing of blood and stain
Moisture in air/poor slide drying
RBC inclusions

70
Q

What does poor mixing of blood and stain do to reticulocyte observation?

A

Retics sit on top of stain giving inaccurate count

71
Q

What does moisture in air or poor slide drying do to reticulocytes on smear?

A

Refractile appearance of cells on slide

72
Q

What other RBC inclusions can affect Retic counts

A

Heinz bodies
Howell Jolly bodies
Pappenheimer bodies

73
Q

What RBC inclusions appear very similar to reticulocytes?

A

Pappenheimer bodies

74
Q

What are stress or shift reticulocytes?

A

Cells younger than “normal” reticulocytes so they are larger and more blue

75
Q

What is the appearance of stress or shift reticulocytes

A

Larger
More blue

76
Q

What is a “normal” hematocrit?

A

45%

77
Q

What is the formula for corrected reticulocytes?

A

(PT Hct /45%) * (Raw Retic %)

78
Q

What is the corrected retic count used for?

A

Interpreting reticulocyte release in response to anemia

79
Q

What is Reticulocyte Production Index (RPI)?

A

Correction that takes into account reticulocytes leaving marrow on a daily basis

80
Q

What is the normal number of maturation days in blood?

A

One day

81
Q

How long does it normally take a reticulocyte to mature in the blood?

A

One day

82
Q

Normal range Hct

A

35-45%

83
Q

For every two percentage point drop in the Hct, the reticulocyte maturation time is increased by how many days?

A

0.1 days

84
Q

How does percentage drop in hct affect the reticulocyte maturation time?

A

For every 2% HCT decrease, the maturation time increases by 0.1 days

85
Q

RPI Formula

A

(PT HCT/45%) * (Raw Retic %/#Days to mature)

86
Q

Number of days to mature calculation

A

(PT HCT - 45%) = x / 2 = y * 0.1 = z + 1

87
Q

Percentage of reticulocytes

A

(# Reticulocytes / 1000 RBCs) * 100 = %
or
(# Retics/10) = %

88
Q

Adult Reticulocyte reference range

A

0.5-2.0%

89
Q

Newborn Reticulocyte reference range

A

2.5-6.0% (up to age two weeks)

90
Q

Up to what age is a newborns reticulocyte reference range going to be different from an adults?

A

Up to two weeks of age

91
Q

What is the formula for absolute reticulocyte count

A

(% Retics) * (# RBCs)

92
Q

What is the absolute reticulocyte count

A

retics per microliter

93
Q

Absolute reticulocyte reference range

A

25,000 - 175,000 reticulocytes/mcL
or
25-175 x 10^9 reticulocytes/L

94
Q

Does this RPI indicate an appropriate response? >3 RPI

A

Yes, this indicates there is an RBC destruction problem and the bone marrow is responding adequately

95
Q

Does this RPI indicate an appropriate response? <2 RPI

A

No, this indicates an inadequate response
Bone marrow production problem

96
Q

How should the absolute retic count look if a person is anemic?

A

Elevated

97
Q

What is the better way to express a patient’s reticulocyte count?

A

Absolute reticulocyte count

98
Q

What is the IRF?

A

Immature reticulocyte fraction

99
Q

How do younger retics stain and why?

A

More densely
have more m-RNA

100
Q

When does IRF increase?

A

BEFORE RPI, Hgb, or Hct when an intervention has worked (EPO therapy, bone marrow transplant)

101
Q

Which is preferred RPI or IRF?

A

IRF

102
Q

Hemoglobin nomenclature on instrument

A

CHr
RET-He

103
Q

What does the hgb content look like in a patient with an iron deficiency?

A

Low hgb content of RBC

104
Q

When a patient is responding to iron therapy, where is this first detected?

A

Hgb levels in retics

105
Q

If 30 retics were counted using. miller disk, what is the % Reticulocytes?

A

3%