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BHCS2005 > methods in haematology II > Flashcards

methods in haematology II Flashcards

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1
Q

Mean Cell Haemoglobin (MCH) =

A

Haemoglobin (Hb) divided by Red cell count (RBC)

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2
Q

Mean Cell Volume (MCV) =

A

Packed cell volume(PCV) divided by Red cell count (RBC)

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3
Q

Mean Cell Haemoglobin

A

Concentration (MCHC) =
Haemoglobin (Hb)divided by Packed cell volume(PCV)

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3
Q

Haematocrit (HCT or PCV packed cell volume)

A

Can be calculated based on red cell count & size OR measured in centrifuged capillary tube.

  • Males 40-52%
  • Females 38-48%
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4
Q

what is haemoglobin typically measured in

A

Haemoglobin is typically measured in aggregate on lysed cells and compared to the red cell number and size]

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5
Q

what is red cell number and volume measured by

A

Red cell number and volume is measured by scatter or impedance

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6
Q
  • Mean corpuscular haemoglobin (MCH)
A
  • Mean corpuscular haemoglobin (MCH) is a calculation of the amount of oxygen-carrying haemoglobin inside your RBCs.
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7
Q

Mean corpuscular volume (MCV)

A

Mean corpuscular volume (MCV) femtolitres calculated from PCV/RBC count or averaging of red cell size

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8
Q

would macrocytic RBC have higher MCH (mean corpuscular haemoglobin)

A

Since macrocytic RBCs are larger than either normal or microcytic RBCs, they would also tend to have higher MCH values.

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9
Q
  • Mean corpuscular haemoglobin concentration (MCHC)
A

is a calculation of the concentration of haemoglobin inside the RBCs.

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10
Q

what is decreased MCHC (mean corpuscular haemoglobin concentration) seen in what conditions

A

Decreased MCHC values (hypochromia) are seen in conditions where the haemoglobin is abnormally diluted inside the red blood cell
cells, such as in iron deficiency anaemia, long standing inflammation or thalassaemia.

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11
Q

increased MCHC mean corpuscular haemogolbin concentration diseases

A

are seen in conditions where the haemoglobin is abnormally concentrated inside the red cells, such as in hereditary or
autoimmune spherocytosis.

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12
Q
  • Red cell distribution width (RDW)
A

) is a calculation of the variation in the size of your RBCs (by impedance or cytometry).
RDW-SD is the standard deviation, RDW-CV is the coefficient of variation (SD/Mean).

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13
Q

Anti inflammatory tests

A

c-reactive protein – good indicator up to 24h in inflammatory response
plasma viscosity and ESR- for monitoring

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14
Q

blood analyser flow cytometry =

A

sample typically separated to measrure RBC/ platelets and WBC.

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15
Q

the blood smear

A

Geimsa stain - Methylene blue, Azure B & Eosin
(Wright stain - Methylene blue & Eosin)

Erythrocytes - pink,
Platelets - light pale pink
Lymphocyte cytoplasm - sky blue
Monocyte cytoplasm - pale blue
Leukocyte nuclear chromatin - magenta
Eosinophil granules - red
Basophil granules - blue/purple

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16
Q

ELIZA enzyme linked immunosrbent assay

A

1) Coated antibody specific for protein masured
2) Plate washed remove excess
3) Plasma is added binds to the immoblised antibody
4) Plate is washed removes plasma and protien
5) Addition of antibody that is conjugated to an enzyme the antibody binds to the protien.
6) Plate is wajsed and rmoved excuesses antioby and fluid
7) Substarte is reacted with the enzyme to produce a colour change.

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17
Q

Assays for clotting disorders

A
  • Prothrombin time (PT): measures extrinsic
    pathway – measures clot formation upon the
    addition of thromboplastin
  • Thrombin Time (TT): measures clot formation
    upon addition of thrombin
  • Fibrinogen: measures adequate levels of
    fibrinogen for clot formation
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18
Q

LIA
Latex immunoassay (LIA)

A

Latex particles coated with antibody of protien being measured , agglutation measured turbidmetrically

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19
Q

Activated partial thromboplastin (APTT) measured the intrinsic pathway

A

which measures clot formation in the presence of APTT reagent and calcium ions

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20
Q

micronutrients (vitamins /Minerals) required for RBC production

A

iron , vitamin B12 , folate , Vitamin B6 , ferritin

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21
Q

the uptake of iron

A

iron is absorbed from the duodenum and jejnum
best absorbed is the Fe2+ form , is it transported in plasma bound to transferrin which delivers itr to tissues these tissues have transferrin receptors, it is stored in tissues bound to ferritin.

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22
Q

what is Hepcidin

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A

(peptide hormone produced by liver) regulates iron uptake from gut by blocking iron export from cell by ferroportin and degrading ferroprotein

22
Q

what is required for DNA synthesis

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A

Vitamin B 12 and folate

23
Red blood cell composition ?
alpha chain , beta chain , heme group beta chain , alpha chain 2 (helical shape of the polypeptide molecule.
24
what is Anaemia
Low red cell number or low levels of Hb <13g/dL (men), <12g/dL (women), <11gdL (pregnant women) Usually a local reference range: 13.3-16.7g/dL men 11.8-14.8g/dL women
25
Abnormal and malignant haematopiesis
haemophilla and clotting disorders VWF and clotting cascade , CML, AML, MPD pathogenesis , MM ALL , CLL pathogens
26
Stem cells
Can self renew Few in number Can differentiate into several lineages One stem cell can produce 1x106 cells after 20 divisions Cannot be distinguished morphologically – requires functional assays Usually located in a protected environment ‘tucked away deep in tissues’ away from immediate harm Hypoxic niches, frequently glycolysis rather than oxidative phosphorylation
27
Haematopoietic stem cells
CD34+ &/or CD133+ (cluster of differentiation marker) c-kit high Sca-1 high Lin- lineage negative i.e. defined by what they don’t express
28
Lin- cocktail different markers
CD3: A marker for T lymphocytes. CD4: A marker for helper T cells. CD8: A marker for cytotoxic T cells. CD19: A marker for B lymphocytes. CD20: Another marker for B lymphocytes. CD14: A marker for monocytes. CD16: A marker for natural killer (NK) cells & some monocytes. CD56: A marker for NK cells
29
Osteoblasts:
factors maintain HSC quiescence (a non-dividing state) and support their self- renewal.
30
Endothelial Cells:
factors that promote HSC proliferation and differentiation.
31
Mesenchymal Stem Cells (MSCs):
source of stromal cells and secrete regulatory molecules that influence HSC behavior. They become osteoblasts (bone-forming cells), chondrocytes (cartilage cells), and adipocytes (fat cells).
32
Macrophages:
IL-6 and TNF-α can promote HSC proliferation, IL-10 can induce quiescence
33
Stem cells:
Pluripotent can renew or differentiate into all haematopoietic lineages Multipotent – can differentiate into multiple lineages Lineage committed can only develop along that ‘line’ of differentiation Fully differentiated cells with function in blood
34
haematopoeisis is required for
transpprt of oxygen fight infection haemostasis (blood clotting) transport of nutrients and removal of toxic products
35
cellular processes invovled
ØProliferation – cell division and growth ØDifferentiation – acquisition of specific maturation characteristics for function ØApoptosis - cell death * Healthy individuals: 5-10 x 1011 blood cells per day
36
Where haemtoposiesis takes place ?
Sternum , pelvis , femurs
37
Normal bone marrow
* Haematopoietic tissue: stem cells and progeny * Mesenchymal stem cells that produce stromal cells and fibroblasts that secrete scaffolding proteins such as collagen * Macrophages: produce growth factors to regulate haematopoiesis * Adipocytes: store energy in form of fat Pluripotent: able to produce progeny of all lineages Oligopotent: able to produce progenitor cells that may differentiate into a few cell types ie lymphoid or myeloid stem cells
38
Myeloblast –
myeloid pathway
39
Lymphoblast –
lymphoid pathway
40
Erythroblast –
erythrocyte pathway
41
Colony Forming Assays
Diagnose leukemia, myelodysplastic syndromes (MDS), aplastic anaemia & monitoring of chemotherapy and transplantation Colony-forming unit–granulocyte-erythrocyte-monocyte-megakaryocyte (CFU- GEMM) Colony-forming unit–lymphocyte (CFU-L) Colony-forming unit–erythrocyte (CFU-E) Colony-forming unit–granulocyte-macrophage (CFU-GM) Colony-forming unit–megakaryocyte (CFU-Meg) Colony-forming unit–basophil (CFU-B) Colony-forming unit–eosinophil (CFU-Eos)
42
growth factors and cytokines
Interleukins (IL) SCF (IL-11) – proliferation and development of pluripotent and progenitor cell development IL-1 stimulates production of GM-CSF, G-CSF, M-CSF and IL-6 IL-3, IL-4 and IL-6 act on early multipotential cells IL-5 is eosinophil CSF Colony Stimulating Factors (CSF) G-CSF stimulates differentiation of granulocyte precursors and activates mature granulocytes GM-CSF stimulates differentiation of granulocyte and macrophages Other growth factors Erythropoietin (EPO) – released from kidney and stimulates red blood cell production in bone marrow Thrombopoietin (TPO) – produced by liver and stimulates megakaryocyte maturation and platelet production in bone marrow
43
IL-1
stimulates production of GM-CSF, G-CSF, M-CSF and IL-6
44
IL-3, IL-4 and IL-6
act on early multipotential cells
45
IL-5
is eosinophil CSF
46
G-CSF
stimulates differentiation of granulocyte precursors and activates mature granulocytes
47
GM-CSF
stimulates differentiation of granulocyte and macrophages
48
Other growth factors Erythropoietin (EPO)
released from kidney and stimulates red blood cell production in bone marrow
49
Thrombopoietin (TPO)
produced by liver and stimulates megakaryocyte maturation and platelet production in bone marrow
50
Erythropoiesis
Life span of RBC is 100-120 days 20 seconds to circulate 2-3 x 1013 RBC normally ¼ of the body’s cells
51
Granulopoiesis
Granulocytes: Neutrophil (3- 5 lobes), Basophil (2 lobes), Eosinophil (2 lobes). Distinct subset of granules containing Bioactive enzymes for function
52
Thrombopoiesis
(platelet production) * Platelets for blood clotting arise from the megakaryocyte (Mk) via the CFU-EMk progenitor * Mk’s are polyploidy – have multiple sets of chromosomes (up to 64N as opposed to 2N diploid) * Thrombopoietin (TPO) essential for Mk growth and development of platelets and platelet function – agregration process
53
Monocytes
* Arise from the Granulocyte Monocyte lineage CFU-GMo * Monocytes pass into circulation then migrate into tissues and differentiate into macrophages, they can then further differentiate into more specialised cells such as Langerhans cells etc * Act as immune-surveillance systems
54
Lymphopoiesis
T cell development CLP’s in BM produce double negative (DN) cells (CD4 and CD8 negative) migrate to thymus, development allows expression of double positive cells (CD4+CD8+) then generating MHC class I restricted CD8+ cytotoxic T cells and helper MHC class II helper T cells. T regs express CD4+ CD25+ B cell development CLP’s generate immature B cells which migrate to lymphoid tissues to encounter antigen. Activated B cells secrete IgM or IgG antibodies or may differentiate into plasma cells

BHCS2005 (3 decks)

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