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Pathophysio > LabD 5 RBC > Flashcards

LabD 5 RBC Flashcards

1
Q

Main function?

A

Oxygen transportation

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2
Q

Problem with RBC causes?

A

Hypoxia in tissues

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3
Q

What can cause disease of RBC?

A

Transitions in RBC metabolism, many diff substances is produced. Transitional products

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4
Q

Most important RBC parameters?

A

RBC count, haemoglobin conc. and or function

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5
Q

RBC count, haemoglobin conc. and or function, these parameters can help us evaluate?

A

Polycytaemias and anaemias caused by diff external and internal causative agent

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6
Q

Haemoglobin measurement; spectrophotometric method (Drabkin method) how?

A
  • Whole blood sample to reagent (potassium ferricyanide) which hemolyses RBCs and forms Fe3+ from Fe2+ in the haemoglobin molecule -> further oxidized by potassiumcyanide to cyanidmethaemoglobin
  • Mix it and measure orange color enproduct by spectrophotometer
  • Use standard solutions or standard curve to compare
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7
Q

Spectrophotometer method (Drabkin method) formula

A

Esample/Estandard x standard concentration = result (in same unit as standard) (E = extinction)

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8
Q

Normal value of haemoglobin in blood

A

18-20 mmol/L or 12-18 g/dl (g%)

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9
Q

Drabkin method; hemolyzed RBCs contains almost the whole amount of haemoglobin in blood, but what and where about the rest?

A
  • Very small amount of free Hb, bound to carrier protein Haptoglobin (acute phase protein from last lab)
  • Therefore no notable increase in Hb conc in case of intravascular hemolysis
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10
Q

Oxygen dissociation curve, left and right shift means?

A
  • Left: decresed temp, decreased 2-3 DPG, decreased H+ conc, CO (increased affinity?)
  • Right: reduced affinity, increased temp, increased 2-3 DPG, increased H+ conc
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11
Q

Oxygen binding capacity of Hb is increased by?

A
  • Decreased 2,3 DPG level in RBCs
  • Decreased pCO2 level in blood (ex. resp alkalosis)
  • Decreased temp of blood (hypothermia)
  • Increased pH of the blood (alkalosis, metabolic or resp)
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12
Q

Oxygen binding capacity of Hb is decreased by?

A
  • Increased 2,3 DPG level in RBCs
  • Increased pCO2 level in blood ( ex. resp acidosis)
  • Increased temp of blood (hyperthermia)
  • Decreased pH of blood (acidosis, respiratory or resp)
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13
Q

What is oxygen saturation?

A

Percentage (proportion) of oxygenated Hb molecules compared to the whole amount of Hb molecules in one unit of blood

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14
Q

Normal oxygen saturation values?

A

Arterial: 95-99%
Venous: 80-90%

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15
Q

Causes of increased Hb conc?

A
  • Usually associated with different types of relative (dehydration) or absolute polycytaemia
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16
Q

Causes of decreased Hb conc?

A
  • Usually associated with relative (hyperhydration) or absolute oligocytaemia
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17
Q

Is Hb conc affected by age of animals?

A
  • Usually not, except swine: young pigs have much lower -Hb conc than older ones
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18
Q

Methaemoglobin?

A
  • Hemoglobin containing Fe3+ –> unable to carry oxygen

- Small amount in blood, possible to reduce them to Hb by the methamoglobin-reductase enzyme

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19
Q

Hemoglobin transports oxygen to the cells and then

A

it is used in the terminal oxidation phase of the metabolic process

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20
Q

What factors can lead to increased methaemoglobin amount in blood?

A
  • Severe oxidative damage tot he RBCs: ex. nitrites, free radicals, paracetamol, onion)
  • Called Methaemoglobinaemia
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21
Q

What species are especially sensitive to ox. damage?

A
  • Hb molecules of cats, newborns and very young animals are sensitive to oxidative damage
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22
Q

Methaemoglobinaemia, colour?

A
  • Colour is dark brown and mucous membranes are deeply cyanotic
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23
Q

Rough estimation formula for Hb conc?

A

PCV (1/) / 3 x 1000 = Hbg (g/L)

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24
Q

RBC count methods?

A
  • Burker chamber….
  • Estimated RBC count; formula used if we suspect a normal average RBC volume
  • RBC count measured by automatic cell counter
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25
Q
  • Estimated RBC count; formula used if we suspect a normal average RBC volume
A

Ht L/L / 5 x 100 = RBC count x 10^12/L

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26
Q
  • RBC count measured by automatic cell counter
A
  • Based on electrical impendance change due to transmission of particles through an aperture -> electric flow -> size of RBC
  • Shows RBC and thrombocytes
  • X axis shows size, Y axis shows number counted
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27
Q

RBC size taken up by automatic cell counter

A

40-100fl (0-150)

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28
Q

Automatic cell counter, why should blood be warmed to 37 C before counting?

A

Because if the blood is cold-agglutinated the cells in the aggregrations will not be counted. Warm -> separates cells

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29
Q

Normal RBC count

A

4,5-8 x 10^12 /L

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30
Q

Tests to gain info about size, number, colour (the indices)

A
  • Ht, PCV, RBC count, Hb concentration (objective info), blood smear if by good cytologist (can be very subjective)
  • They are useful for humans, cats and dogs, but for horses and cattle the parameters of RBCs are varying too much, also within the individual
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31
Q

Mean corpusclar Haemoglobin (MHC) indicates?

A

MCH indicates average Hb content of RGCs

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32
Q

Mean corpusclar Haemoglobin (MHC) formula

A

Hb (g/L) / RBC count x 10^12/L = MHC (pg)

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33
Q

Mean corpusclar Haemoglobin (MHC), normal value?

A

12-30 pg
In young animals it can be increased 28-31 (MCV can also be increased)

Horse - 12-20
Ru - 8-17
Dog - 15-24
Cat - 13-17

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34
Q

Decreased MCH is called?

A

Hypochromasia

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35
Q

Increased MCH is called?

A

Hyperchromasia

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36
Q

Mean corpusclar volume (MCV) indicates?

A

MCV indicates the average size of the RBCs (macro, normo, microcytic)

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37
Q

Mean corpusclar volume (MCV) formula?

A

PCV / RBC count x 1000 = MCV (fl)

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38
Q

Mean corpusclar volume (MCV) normal values?

A 
60-70 fl
Horse - 37-58
Ru - 42-52
Dog - 63-75
Cat - 40-53
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39
Q

MCV, size og RBC in species, age, and some breeds

A
  • Great heterogenity in size
  • Cats, horses -> smaller RBC
  • Young RBC -> large
  • Newborns -> large
  • Adults -> smaller
  • Japanese Akita -> smaller
  • Poodles -> very large
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40
Q

Some causes of microcytosis:

A
  • Chronic blood loss
  • Iron, copper, pyridoxine (vi. B6) deficiency
  • Portosystemic shunt
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41
Q

Some causes of macrocytosis:

A
  • Mostly regenerative anemias
  • Polycythaemia absoluta vera (erythroleukemia)
  • Vit. B12, folic acid, cobalt deficiency
  • Erythroleukemias
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42
Q

Mean corpusclar Haemoglobin concentration (MCHC) indicates?

A

MCHC indicates the average concentration of haemoglobin in erythrocytes (hB concentration)
(hyper, normo, hypochromic RBCs)

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43
Q

Mean corpusclar Haemoglobin concentration (MCHC) formula?

A

Hb (g/L) / PCV = MCH (pg) / MCV (fl) x1000 = MCHC (g/L)

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44
Q

Mean corpusclar Haemoglobin concentration (MCHC) normal values?

A

300-350 g/L (30-35%) - normochrom

Horse - 31-37 %
Ru - 30-36 %
Dog - 32-36 %
Cat - 30-36 %

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45
Q

When MCV and MHC are low (ex. chronic liver failure) MCHC can be normal, however there is?

A

Anemic state

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46
Q

Decreased MCHC- hypochromasia causes:

A
  • Newborn animals
  • Regenerative anemias
  • Iron deficiency anemia
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47
Q

Increased MCHC- hypochromasia causes:

A
  • Erythroleukemia (polycythaemia absoluta vera)
  • Vitamin B12, folic acid, cobalt deficiency
  • Immunohemolytic anemia (spherocytosis-presence of blood in spherocytes)
  • Lead poisoning
  • Splenectomy
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48
Q

Increased MCHC- hyperchromasia causes:

A
  • Erythroleukemia (polycythaemia absoluta vera)
  • Vitamin B12, folic acid, cobalt deficiency
  • Immunohemolytic anemia (spherocytosis-presence of blood in spherocytes)
  • Lead poisoning
  • Splenectomy
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49
Q

Typical changes in derivated parameter:

macrocytic, hypochromic; MCV-up, MCHC-down, (reticulocytes-up)

A

Regenerative anemias

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50
Q

Typical changes in derivated parameters: normocytic, normochromic; MCV-unchanged, MCHC-unchanged, MCH normal or decreased

A

Non regenerative anemias

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51
Q

Typical changes in derivated parameters: microcytic, hypochromic; MCV-down, MCHC-down, (decreased Hb synthesis)

A

Iron, copper, piridoxine deficiency anemias, liver failure, portosystemic shunt

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52
Q

Typical changes in derivated parameters: microcytic, normochromic; MCV-down, MCHC-unchanged

A

Normal for Japanese Akita

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53
Q

Typical changes in derivated parameters: macrocytic, normochromic; MCV-up, MCHC-unchanged, impaired DNA synthesis

A

FeLV infection, vitamin B12, CO or folic acid deficiency, erythroleukemia, poodle macrocytosis

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54
Q

Red Cell Distribution width, Platelet Distribution Width (PWD) are indices that gives us info about?

A

They give a number that is correlated with the range of the average size of the RBCs and platelets

55
Q

Where is RDWs width on the graph?

A

It is the line between the two points (P1-P2) where the horizontal line crosses the two sidelines of the curve. We express it in the percentage of P1-P2 width compared to the distribution of all RBC (from 0-approx100 fl) as 100%

56
Q

RDW dog and cat:

A

Dog 12-16%
Cat 14-18%
Normally the histogram is symmetric Gauss-curve with slight right shift

57
Q

PDW dog and cat:

A

Dog 6-8%
Cat 7-12%
Normally the histogram is symmetric Gauss-curve with slight right shift

58
Q

Short RDW means?

A

non regenerative processes

59
Q

Large RDW means?

A

regenerative processes

60
Q

What are reticulocytes

A

Young, but differentiated RBCs with basophil punctates stained by Brylliant-cresis blue stain. Blue punctates are rRNA remnants.

61
Q

Reticulocytes containing big blue aggregates (aggregated forms) are?

A

Younger forms

62
Q

Reticulocytes containing small punctates (punctated forms) are?

A

Older forms. In cats punctated forms are more normal.

63
Q

No reitculocytes in what species?

A

Horses and ruminants. They exist only in bone marrow, not in the peripheral blood

64
Q

Appearance of reticulocytes are a sign of?

A

regenerative function of the bone marrow

65
Q

Does reticulocytes have the same functional properties as mature RBCs?

66
Q

Does reticulocytes have the same functional properties as mature RBCs?

A

yes, able to carry oxygen

67
Q

Why do we count reticulocytes and not nucleated RBCs when differentiating between regenerative and non-regenerative anemias?

A

Because in case of a maturation arrest of RBCs, nucleated RBCs will never become reticulocytes or mature RBCs. Nucleated RBCs are too young to carry oxygen, reticulocytes are

68
Q

What can cause maturation arrest of nucleated RBCs?

A

ex. vitamin B12 or folic acid deficiency, or FeLV

69
Q

Regenerative anemias are ususally diseases with favourable prognosis, because?

A

Enough new RBCs are produced in the bone marrow to regenerate the anemia, to replace the lost RBCs and to reach normal RBC count quickly.

70
Q

If the bone marrow function is normal, RBC production os ?

A

In positive correlation with the severity of anemia

71
Q

In case of severe anemia more intensive RBC production (a more significant increase in reticulocyte count) is needed to?

A

consider this as a regenerative anemia, than in case of mild anemia

72
Q

Reticulocyte staining

A
  • Brylliant-cresil blue in physiological saline, then add Na-citrate in solution, mix.
  • Vital staining: mix fresh EDTA blood and Brylliant-cresil in same proportion, (stain fresh/alive/vital/RBCs)
  • Incubate in room temp, take up stain, prepare smear
73
Q

Reticulocyte counting

A
  • Count 100-1000 RBCs and take the percent of the reticulocytes
74
Q

Normal reticulocyte count

75
Q

In order to say that the bone marrow is capable fro replacing the red blood cells in an organism which lost RBCs, we expect that the more severe anemia will cause?

A

increased reticulocytic %. The possible PCV changes we can expect a specific reticulocyte %. The expected values for a proper regenration can be read by table in lab

76
Q

Corrected reticulocyte count (CRC) formula

A

CRC = reticulocyte % x RBC count

77
Q

Normal reticulocyte count?

A

<0,06 x 10^12/L - without anemia

78
Q

Corrected reticulocyte percentage (CRP) formula

A

CRP = Ht patient / Ht average (0,45 dog - 0,37 cat) x reticulocyte %

79
Q

Normal corrected reticulocyte percentage (CRP)

A

<1-2% - without anemia

80
Q

Increased reticulocyte count can be caused by diff types of regenerative anemias:

A
  • acute blood loss (approx 3-5 days are needed for the bone marrow to increase the reticulocyte countin the blood)
  • hamolytic anemia
  • chronic blood loss
  • some types of nutrient deficiency anemias
81
Q

Osmotic resistance of RBCs is dependent on?

A

dependent on the pH of the plasma (normal is 7,4), the reagents , temperature, osmotic conc of plasma and the reagents (NaCl conc), RBC membrane status, regenerative status (reticulocytes are more resistant), HbF (fetal Hb) content of RBCs. (fetal RBCs containing HbF are more resistant)

82
Q

What test can we perform to examine the RBC membrane function? (membrane integrity)

A

Osmotic resistance analysis

83
Q

Normal causes of membrane damage

A
  • Nephropathy (uremia)
  • Specific membrane damage (immunohemolytic anemia)
  • Increased physical damage (long term severe physical activity, ex. long distance running)
84
Q

In general, osmotic resistance becomes decreased in case of?

A
  • Chronic hemolytic anemia types, where there is extracellular hemolysis and the plasma color does not reflect to the hemolytic process
  • > Chronic immunemediated hemolytic anemia is typical for this kind of RBC damage
  • There are some rare hereditary genetical defects causing decreased osmotic resistance (therefore decreased lifespan) of RBCs: pyruvate kinase or glucose 6 phosphate dehydrogenase deficiency in dogs, methaemoglobin reductase deficiency in dogs and horses etc.
85
Q

Method 1, osmotic resistance of RBCs

A
  • Dilution line of NaCl from 0,3 % to 2,5%
  • Drip blood samples into the saline test tubes, incubate in room temp for 10 min
  • Centrifuge -> check upper layer for hemolysis
  • Distilled water with pH around 7.4
86
Q

Method 2, osmotic resistance of RBCs

A
  • Hypotonic solution of NaCl
  • Prepare three tubes; Tube 1 - physiological saline for blood sample of the sick animal, Tube 2 - hypotonic NaCl solution for blood sample of sick animal, Tube 3 - hypotonic NaCl for blood sample of control healthy animal
    -> 5 drops of blood in each tube, incubate 10 min, centrifuge, check upper layer
    Result: if the upper layer of tube 3 shows hemolysis, repeat analysis (analytical error)
    Tube 1,2 - intravasal hemolytic crisis ex. babesiosis, immunehemolytic anemia
87
Q

Erythrocyte morphology; correct blood smear analysis begins with?

A
  • Using proper staining methods, ex. May-Grunwald, Giemsa, Diff quick ex.
  • Smears must be prepared by using fresh samples
88
Q

Erythrocyte morphology; correct blood smear analysis

A
  • Staining method
  • Fresh sample
  • Check blood films on low then high magnification (200-1000x)
  • Check gross signs: rouleau formation - coin arrangement, RBC aggregates, large cells (horse often), thrombocyte aggregates
89
Q

Gross signs in smear: rouleau formation - coin arrangement, what species?

A

Horse often, dog, cat, swine sometimes, cattle rare

90
Q

Intensity of staining RBCs; polychromasia, hyperchromasia

A

More intensive staining

RNA, or nuclear remnants, more Hb - regenerative process

91
Q

Intensity of staining RBCs; hypochromasia

A

Weak staining

Decreased Hb content, iron or other nutrient deficiency

92
Q

Size of RBCs; macrocytosis

A

many big cells

93
Q

Size of RBCs; microcytosis

A

many small cells

94
Q

Size of RBCs; anisocytosis

A

variable cell size, iron deficiency and regenerative process

95
Q

Size of RBCs; poikylocytosis

A

variable size and colour

96
Q

RBC types: young and nucleated RBCs (in order of maturation)

A
  • Proerythroblast
  • Basophil erythroblast (normocyte, normoblast)
  • Polychromatophil erythroblast (normocyte, normoblast)
  • Acidophil erythroblast (normocyte, normoblast)
97
Q

RBC types: young but mature RBC without nucleus

A

Reticulocyte

98
Q

RBC types: Appearance of young RBCs

A

Increased production (regenerative anemia), spleen or bone marrow disease, leukemia, extramedullar erythrocyte procuction, Pb toxicosis (with basophil punctates), hyperadrenocorticism

99
Q

RBC types: reticulocyte appearance

A

Increased production (regenerative anemia) - chronic Fe deficiency anemia, hemolysis, acute blodd loss, chronic blood loss

100
Q

RBC types: Spherocyte (spherical small polychromatophil RBC) appearance

A

sensitive RBC membrane, immunemediated hemolysis

101
Q

RBC types: Stomatocyte (mouth shaped RBC) appearance

A

increased RBC production (regenerative anemia

102
Q

RBC types: Acantocyte (spur cell - RBC with few long spikes) appearance

A

RBC membrane failure (lipid bilayer) - lipid metabolism disorder, hepatopathies

103
Q

RBC types: Schysocyte (RBC fragment) appearance

A

traumatic or toxic damage (uremia, blood parasites, long term severe physical activity, DIC)

104
Q

RBC types: Anulocyte (0-like RBC) appearance

A

iron deficiency anemia

105
Q

RBC types: Codocyte (target cell, like a target) appearance

A

regenerative process

106
Q

RBC types: Echynocyte (Burr cell, crenation, RBC with many small spikes) appearance

A

laboratory error, too quick drying of blood film, uremia, DIC

107
Q

RBC types: Sickle cell appearance

A

RBC damage, Hb globin chain malformation in humans

108
Q

Inclusion bodies in RBCs: Heinz body (NMB - new methylene blue stain); denatured Hb appearance

A

02 effect
Oxidative damage to RBCs (Cat: ex. methaemoglobinaemia)
GSH deficiency

109
Q

Inclusion bodies in RBCs: Howell-Jolly body; nuclear remnants appearance

A

vitamin B12 deficiency
increased prod of red cells
splenectomy

110
Q

Inclusion bodies in RBCs: Basophilic punctuates; nuclear remnants appearance

A

regenerative process
young RBCs of cat
physiological in Ru
lead poisoning

111
Q

Inclusion bodies in RBCs: Hb inclusions appearance

A

Hb damage
increased RBC production
regenerative anemia

112
Q

RBC parasites!

A
  • Haemobartonella canis, felis, bovis
  • Babesia spp., B. canis very common in Hu
  • Ehrlichia canis, equi etc.
  • Dirofilaria immitis, repens
  • Anaplasma marginale, centrale, ovis
  • Eperythrozoon wenyoni, ovis, suis, parvum
  • Citauxzoon felis
  • Theileria parva, mutans, annulata, hirci, ovis
  • Trypanosoma cruzi, congolense, vivax, brucei, evans, suis, equiperdum
  • Leishmania donovani
113
Q

Other Laboratory measurements in connection with RBCs:

A
  • Serum iron measurement
  • ## Total iron binding capacity (TIBC)
114
Q

Serum iron measurement, why and what

A

If we suspect iron deficiency, especially due to chronic blood loss, we can prove it by performing this test

115
Q

Serum iron measurement; normal values

A

Normal SeFe (serum iron): 18-20 umol/L

116
Q

Serum iron measurement, method

A
  • Serum sample is used, fibrinogen of plasma can disturb the measurement.
  • Fe3+ is reduced to Fe2+ by ascorbic acid.
  • Fe2+ reacts with ferrosin and forms a red coloured chelate (complex molecule) which can be measured photometrically
117
Q

Causes of high serum iron conc:

A

Iron toxicosis (overload)

118
Q

Serum iron measurement should always be performed together with?

A

TIBC analysis (total iron binding capacity)

119
Q

Total iron binding capacity (TIBC), what and why

A

This test gives information about the transferrin content.

120
Q

TIBC normal values:

A

50-68 umol/L

121
Q

Blood needed for serum iron measurement and TIBC?

A

Serum iron measurement: serum, fibrinogen of plasma may disturb measurement
TIBC:

122
Q

TIBC normal values:

A

50-68 umol/L,

normal values of formula: 20-55% (30%)

123
Q

TIBC, method

A

Measure seFe, add Fe-solution to the plasma (all transferring molecules will be fully saturated), put absorbent to the solution, centrifuge -> absorbent binds to free Fe and goes to sediment. Use upper layer and check seFe again.

124
Q

Causes of low TIBC:

A
  • chronic inflammation (negatove acute phase proteins)
  • chronic liver failure (decreased transferrin synthesis in liver
  • neoplastic disease
125
Q

Causes of high TIBC:

A

iron deficiency anemia (not severe: normal iron level + high TIBC, severe: low iron level + high TIBC)

126
Q

TIBC: iron saturation formula

A

SeFe / TIBC x 100 = iron saturation

127
Q

Laboratory findings in Hemolysis:

A 
PCV - down
Reticulocytes - up (regenerative anemia)
Polychromasia, poikilocytosis
Leukocytosis, (neutrophilia)
Spherocytosis
Total bilirubin - up
Indirect bilirubin - up
Lactate dehydrogenase 1, 2 - up
Haptoglobin - down
RBC osmotic resistance - down
Jaundice 
Hyperchromic stool
Urobilinogen and Hgb in urine - up
128
Q

Ferritin, RIA method normal values:

A

12-300 ug/L

129
Q

Transferring, method:

A

RIA, ELISA

130
Q

RBC lifespan, method:

A

Cr51 method

131
Q

Hgb electrophoresis, method

A

globin chain sequence analysis

132
Q

Vitamin B12 measurements

A

RIA method

133
Q

Erythropoietin, method

A

ELISA method, highly species specific , a few laboratories can measure it

134
Q

TIBC =

A

serum iron level (saturated transfering) + free transferrin (not saturated)

Pathophysio (12 decks)

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