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Hematology/Oncology > Structure and Function - Lecture 1 > Flashcards

Structure and Function - Lecture 1 Flashcards

1
Q

What are the steps to looking at blood cells with a microscope?

A

Get some blood, don’t let it clot (use EDTA - lavendar), put drop on a slide, use another slide to spread the blood out, let it dry, stain it with “wright giemsa” stain

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2
Q

Describe the eosin stain

A

negatively charged and aromatic (acidic), soluble in ethanol but not water - why you have to let the slide dry before adding

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3
Q

what does eosin stain

A

hydrophobic basic macromolecules like hemoglobin

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4
Q

what is the charge of the heme group

A

positive (aromatic)

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5
Q

what do fluroescin isothiocyanate FITCs bind

A

amines

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6
Q

what percent of leukocytes are eosionphils

A

less than 5 % (rare in blood)

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7
Q

what do eosinophils do

A

mediators of innate immunity

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8
Q

when do you see a lot of eosinophils

A

parasite infection or allergic reaction or as a part of neoplastic malignancy

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9
Q

what is the starting point for diagnosing any increase in any type of blood cell

A

reactive vs neoplastic

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10
Q

what does methylene blue stain well

A

nucleic acids and some protein species

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11
Q

what are the properties of methylene blue

A

flat and positively charged, aromatic and basic

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12
Q

what is methylene blue soluble in

A

water or methanol

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13
Q

what does methylene blue stain

A

hydrophobic acidic macromolcules

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14
Q

what percent of leukocytes are basophils

A

less than 1 percent (rare)

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15
Q

what are the functions of basophils

A

degranulation in allergic reactions (related to tissue mast cells); increase in important neoplastic events; only rarely increase in non neoplastic events

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16
Q

What cell types have few or no cytoplasmic granules

A

monocytes and lymphocytes

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17
Q

what percent of leukocytes are monocytes

A

3 to 8 percent

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18
Q

what is the role of monocytes

A

phagocytes which present foreign antigen via MHC class 2

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19
Q

where do macrophages derive from

A

most (but not all) derive from monocytes

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20
Q

what does monocytosis signify

A

nothing - not specific at all

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21
Q

what cells do monocytes share info with and where

A

lymphocytes in the lymphatic tissues

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22
Q

what percentage of leukocytes are lymphocytes

A

20-30%

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23
Q

when do you see an increase in lympochytes in the blood

A

neoplastic events (leukemia) or viral syndromes

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24
Q

how long do lymphocytes live

A

long, days to years

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25
Q

what are the predominant lymphocytes

A

t cells then b cells then nk cells

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26
Q

what is a reactive lymphocyte

A

has a nucleus that kind of looks like a monocyte - if you cant tell then call it a mononuclear cell - increase in nmber in viral syndromes

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27
Q

what are large granular lymphocytes

A

NK cells and cytotoxic t cells sometimes show basophilic cyotplasmic granules

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28
Q

what is the first differential diagnosis in patient with reactive lymphocytes

A

viral syndrome

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29
Q

how od neutrophils stain

A

very lightly in cytoplasm with both eosin and methylene blue

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30
Q

what do neutrophils nuclei look like

A

segmented

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31
Q

what is lifespan of a neutrophil

A

less than 1 day

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32
Q

what percent of leukocytes are neutrophils

A

40-70%

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33
Q

when do neutrophils numbers increase

A

10 fold in bacterial infections

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34
Q

what are neutrophils weapons

A

phagocytosis, degranulation, formation of NETS

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35
Q

what is a downside to neutrophils NETS

A

sepsis associated conditions like pre-eclampsia

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36
Q

what is essential in evaluating any infectious disease

A

knowing the neutrophil count

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37
Q

what are the neutrophil requirements

A

migrate in response to chemokines, recognize and swarm the enemy, degrade and immobilize the enemy, and minimize collateral damage

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38
Q

what is leukocyte adhesion defect

A

deficiency in CD18

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39
Q

what does a def in cd18 cause

A

leukocyte adhesion defect

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40
Q

what is CXCR2

A

the il-8 receptor on neutrophils that allows the neutrophil to migrate in response to il-8

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41
Q

what can failure of cytoskeleton to response during adhesion result in

A

immunodeficiency

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42
Q

what is the wiskott aldrich syndrome

A

most of problems in t cells but some in neutrophils; have trouble initiating cytoskeletal reorganization via actin polymerization in response to signals from the cell surface

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43
Q

what are the mechanisms that enable neutrophils to migrate in response to chemokines

A

il-8 receptor, grab and hold devices: integrins (cd11a/cd18 complex), very active cytoskeletons

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44
Q

what is excess chemokine secretion associated with

A

excess inflammation

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45
Q

how do neutrophils recognize and swarm the enemy

A

toll like receptors (original equipment), complement receptors (old), Fc receptors (latest technology); secrete more chemokines: CXCL2, IL-8, TNF

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46
Q

How do neutrophils degrade and immobilize the enemy

A

phagocytose it via same receptors that recognized it; chew up polysaccharide cell wall with lysozyme; chew up proteins with various proteases; kill it with bleach (hypochlorite)

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47
Q

how do neutrophils bleach

A

with myeloperoxidase (mpo)

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48
Q

what do defects in the hypochlorite generating system result in

A

neutrophils can recognize but not kill; granuloma formation - chronic granulomatous disease

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49
Q

what do neutrophils do when they have accumulated and done their job

A

apoptose to minimize excess damage

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50
Q

what happens in neutrophils do not apoptose

A

chronic inflammatory conditions

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51
Q

how much space in bone marrow does neutrophils production take up

A

2/3

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52
Q

what are the percentages of what left shift could mean

A

19/20 - reactive (bacterial)

1/20 - neoplastic

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53
Q

what are immature neutrophils

A 
non segmented nuclei (collectively called left shift)
horshoeshaped nuclei = bands
bean shaped (less mature) = metamyelocytes
round nuclei = myelocytes
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54
Q

what are primary granules in neutrophils

A

seen in myeloid precursors in bone marrow - blue

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55
Q

what are secondary granules in neutrophils

A

pink ones

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56
Q

what does toxic granulation mean

A

increased cytoplasmic granules in the neutrophils - suggests bacterial infectio

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57
Q

what are platelets

A

tiny anucleate fragments

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58
Q

compare platelets in blood to white cells

A

high concentration of platelets (100 times more than white cells)

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59
Q

when does platelet count tend to increase

A

iron deficient patients

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60
Q

what is the lifespan of platelets

A

9-10 days

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61
Q

what are the functions of platelets

A

primary hemostatic plug, stimulate coagulation cascade, stimulate wound healing, immune function

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62
Q

what is the primary hemostatic plug function of platelets

A

adherence, activation, aggregation

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63
Q

what is involved in platelets stimulating coag cascade

A

fibrin formation, clot retraction

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64
Q

how do platelets stimulate wound healing

A

fibroblast growth and migration

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65
Q

how do platelets provide immune function

A

antigen presentation and pathogen inactivation

66
Q

what is the mean of the broad normal range of platelet concentration

A

300,000 per microliter of blood

67
Q

what is the average normal blood volumer

A

4.5 L

68
Q

what is the normal number of platelets in a person with 4.5 L of blood

A

over a trillion

69
Q

what cytokine is secreted by platelets

A

platelet factor 4 PF4 that actually kills the pathogen that causes malaria

70
Q

what are the platelet requirements

A

must be present in large numbers, need to recognize damaged blood vessels, need to swarm onto the damage fast, patch the damage, rebuild the vessel wall

71
Q

how many platelets do you need in order to not bleed spontaneoulsy

A

10,000 per microliter

72
Q

how many platelts do you need to stop small bleeds

A

50,000 per microliter

73
Q

where do you store platelets and when are they released

A

spleen, when you are scared

74
Q

how much can epinephrine increase platelet count by

A

50%

75
Q

what percent of platelets die without doing anything

A

90

76
Q

What else do platelets express that is weird

A

FcR

77
Q

what is the adaptor

A

von willebrand factor

78
Q

how do platelets bind subendothelial collagen

A

surface protein called Gp1b

79
Q

describe how vWF works

A

sticks to exposed collagen and changes shape slightly; flaps in the breeze - platelets bind to it

80
Q

what happens if vWF is in short supply

A

patient bleeds a lot

81
Q

what happens if vWF is too short

A

patients bleed alot

82
Q

what happens if vWF is too long

A

clot excessively

83
Q

where are reserve supplies of vWF

A

plasma, endothelial cells and platelets

84
Q

describe VWF

A

homo polymer

85
Q

what is VWF disease

A

excessive bleeding

86
Q

what causes platelet activation

A

binding collagen or VWF

87
Q

what do activated platelets do

A

bind VWF and fibrinogen more actively and cause aggregation via crosslinking of platelets

88
Q

what is fibrinogen the precursor to

A

fibrin

89
Q

what does autocatalytic mean

A

activation/aggregation is autocatalytic; activated platelets secrete mediators that augment activation/aggregation

90
Q

what does activation and aggregation also promote

A

coagulation cascade

91
Q

what is the coagulation casacde

A

series of plasma proteases that when activated deposits a fibrous mesh made up of mostly fibrin

92
Q

if the coag cascade gets activated first what happens

A

it causes platelet aggregation

93
Q

what is the first differential diagnosis for a patient with broad spectrum reduced production of coagulation factors

A

liver disease since many of them are made in the liver

94
Q

what do activated platelets secrete that stimulate fibroblasts and smooth muscle cells

A

PDGF, TGFbeta, and VEGF

95
Q

when are giant platelets seen

A

peripheral blood when platelet production is ramped up or when production occurs abnormally with a disease affecting bone marrow

96
Q

what are the requirements for red blood cells

A

flexibile shape, durable membrane, maximum Hgb/O2 carrying capacity, offset osmotic pressure, anti-oxidant system, energy supply, ability to tune down complement

97
Q

describe the flexibile shape of red blood cells

A

biconcave disk; 360 deg fold capacity

98
Q

what happens if main cargo of RBC (hemoglobin) precipitates

A

impaired RBC, obstructed blood vessel, ruptured red cell

99
Q

how long do RBC last

A

100 days

100
Q

mutations in rbc shape to give ellipsoid or spherical shape do what to the durability

A

reduce it

101
Q

why is it so easy for hemoglobin to precipitate

A

b/c hemoglobin is packed at the max highest concentration it can be at before it precipitates

102
Q

what can cause hemoglobin to precipitate

A

minor changes in globin amino acid sequence or oxidative stress

103
Q

what is a good bacterial growth medium

A

hemoglobin

104
Q

what is the most important microorganisms that grows on hemoglobin

A

plamodum family of protozoa (4 species cause malaria)

105
Q

what are some mutations that select against infection with bacteria that grow on hemoglobin

A

disabling red blood cells in stress

106
Q

what is the consequence of getting rid of things like nuclei in rbc

A

limits response to changing conditions and pathogens

107
Q

describe the rbc pump system

A

use atp to pump out sodium and in potassium

108
Q

what happens if the RBC pump fails

A

cell swells and bursts

109
Q

what can cause the rbc pump to fail

A

mutations that impair production of RBC

110
Q

what are concentrations of sodium and potassium in the plasma

A

sodium 140 - potassium 4

111
Q

what are the concentrations of sodium and potassium in the rbc

A

sodium 11 - potassium 100

112
Q

why is an anti oxidant system needed

A

o2 is powerful oxidant (via generation of h2o2) - oxidized-sh groups on hemoglobin become crosslinks (R1-SS-R2) which dentaures Hgb or causes it to precipitate and oxidized iron (Fe3+) cant carry O2 which results in hemoglobin containing Fe 3+ (methemoglobin) and patient is hypoxic

113
Q

how does the antioxidant system work

A

GSH eliminates peroxide before it can do damage by using NADPH; cytochrome b5 reductase reduces methemoglobin back to normal hemoglobin using NADH

114
Q

what happens when you have mutations in the anti oxidant system (energy supply) and you are in an oxidatively stressful state

A

clumps of oxidized hemoglobin; can lyse (hemolytic anemia)

115
Q

what is hemolytic anemia

A

when excess red cell lysis occurs

116
Q

how do macrophages help in hemoglobin mutation situations

A

take big bites out of rbc and remove hemoglobin - bite cells

117
Q

describe the energy supply system of the RBC

A

simple, don’t take up cargo space; chuck mitochondria, use glycolysis to make atp and nadh; use pentose shunt to make nadph

118
Q

what is the first potential failure point in the energy supply making of RBC

A

Glucose 6 phosphate dehydrogenase

119
Q

what resistance can happen with G6PD mutations

A

malaria resistance

120
Q

why do we need to tune down complement fixation

A

alternative complement fixation pathway is active at low rate all the time; normal blood cells have to slow it down to survive

121
Q

what does RBC use to slow down complement fixation

A

DAF - CD55; which binds the c3 convertase c4bc2a and does not allow alternative pathway to continue

122
Q

what can not enough hemoglobin mean

A

hypochromic and/or microcytic

123
Q

what does hypochromic mean

A

lack of color

124
Q

what does anisocytosis mean

A

refers to abnormal distribution of red cell sizes

125
Q

what does poikilocytosis mean

A

abnormal red cell shapes

126
Q

what does bluish rbc mean

A

making a lot of residual mrna which means making a lot of cells (good) bc losing a lot (bad)

127
Q

what are reticulocytes

A

bigger than normal just out of bone marrow RBC

128
Q

when are rbc polychromatic

A

usually with accelerated production

129
Q

what can cause precipitated hemoglobin

A

genetic defect in hemoglobin, altered shape (sickle cell); oxidized hemoglobin

130
Q

what are Heinz bodies

A

small clumps of oxidized hemoglobin

131
Q

what causes sickle cell

A

substitution in 6th amino acid in beta globin

132
Q

what 2 diseases give heterozygotes the malaria advantage

A

g6pd mutation and sickle cell

133
Q

what are shistocytes

A

red cell fragments

134
Q

what causes shistocytes

A

mechanical lysis or microangioplastic processes

135
Q

how is hemoglobin measured

A

spectrophotometry; lyse cells and add cyanide (blue color) and quantify non blue light

136
Q

what can mess up the hemoglobin measurement

A

excess lipoproteins after a fatty meal which gives a false high level of hemoglobin measurement

137
Q

how are RBC measured

A

manual methods; centrifuge anticoag blood in tube and spin down cell; sometimes capillary tube; also by conductivity in coulter chamber

138
Q

what is hematocrit

A 
A= red cells B = blood volume 
hematocrit = A/B
139
Q

describe the coulter chamber

A

red blood cells pumped through large container through aperture to smaller container with same fluid as large container; electrode 1 in large electrode 2 in smaller container; voltage gradient applied and resistance across aperture measured

140
Q

what happens to the resistance of the aperture as cells move from large to small container

A

increases; increase directly proportional to cell size

141
Q

how is the red cell count calculated by conductivity

A

flow rate into outer container and resistance spikes per second

142
Q

what is the size of each cell proportional to in the coulter chamber

A

peak height

143
Q

how can we get mean cell volume

A

by peak height

144
Q

how do you get hematocrit

A

multiply number of cells times MCV

145
Q

what could fragmented rbc be counted as

A

platelets

146
Q

what can clumped red cells (maybe by antibodies?) do to hematocrit?

A

lower it

147
Q

what can clumped red cells (maybe by antibodies?) do to red cell count?

A

lower it

148
Q

what can clumped red cells (maybe by antibodies?) do to mcv?

A

increase

149
Q

how can leukocytes be counted

A

manually or automated (by coulter chamber)

150
Q

what other info can be found using coulyter chamber for leukocytes

A

what’s inside cells by measuring resistance to radio freq alt current

151
Q

what is formula for peak height for rbc

A

peak height = k * volume

152
Q

what is formula for peak number for rbc

A

peak number = k * cell count

153
Q

what is formula for peak height for leukocytes

A

peak height = k *cell complexity

154
Q

what cells are hard to differentiate between in coulter chamber

A

basophils and eosinophils

155
Q

what is the flow cytometer used for

A

hit cells with laser and get side scatter measurement

156
Q

how do you get reticulocyte count

A

red cells + nucelic acid binding fluorescent dyes -> flow cytometer

157
Q

what are benefits of reticulocyte count with flow cytometer

A

assess whether anemia is due to impaired red cell production

158
Q

what does the hematology analyzer not reliably count

A

band and other immature granulocytes are counted as neutrophils; blasts counted as lymphocytes of monocytes; red cell fragments may not be detected or counted as platelets; platelet clumps not counted and may show false thrombocytopenia

159
Q

what do platelets clump when sampling sometimes; how can this be fixed

A

presence of EDTA may expose antigens; sample with citrate

160
Q

what is an important differential diagnosis for thrombocytopenia

A

lab artifact

161
Q

what are blasts

A

early blood cell precursors; can correlate with bone marrow diseases such as acute leukemia

Hematology/Oncology (4 decks)

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