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Lab D > Lab 3 - HAEMOSTASIS > Flashcards

Lab 3 - HAEMOSTASIS Flashcards

1
Q

What does haemostasis mean

A

Hemostasis is the name of a group of processes initiated in the body in order to stop bleeding in case of tissue and/or blood vessel injuries.

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2
Q

What are the Major groups of hemostasis disorders.

A
  1. Vasculopathy
  2. Thrombocytopathy
  3. Coagulopathy
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3
Q

What is Vasculopathy

A
  • *The first step** of the hemostasis process
  • *Decreased ability** of vasoconstriction in case of blood vessel injury
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4
Q

What is Thrombocytopathy

A
  • *The second step** of haemostasis
  • *Decreased ability** of platelets to aggregate and adhere to the site of injury, and formation of the primary thrombocyte-thrombus,
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5
Q

What is Thrombocytopenia

A

Decreased amount of thrombocytes in the blood

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6
Q

What is coagulopathy

A

The third and final step of haemostasis

Problems with the extrinsic-, intrinsic-, or common pathway of the coagulation cascade, which
ends with the formation of a polymerized fibrin network, which keeps thrombocyte thrombi at the site of injury,

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7
Q

Which test are performed by the side of the animal

A

1 Signs of increased bleeding tendency:

2 Capillary resistance (human medicine)

3 Bleeding time
(buccal mucosal bleeding time test, BMBTT)

4 The appearance of the first fibrin strand (clotting time)

5 The appearance of the clot (clotting time on different surfaces)

6 Clot retraction time

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8
Q

Where do we look for signs of increased bleeding tendency

A

 Signs of increased bleeding tendency:

  • on the skin and mucous membranes:
  • *anemia**, petechia, ecchymosis,

suffusion;

in the thoracic cavity: haemothorax;
in the abdominal cavity: hemoperitoneum;
in the gastrointestinal tract: haematemesis, melena

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9
Q

What do you know about the Capillary resistance test

A
  • The test is usually used in human medicine,
  • Also called Rumpel-Leed-test.
  • Ligarture on arm
  • Check for PETECHIA on palmar side

3- 5 min

Tells us about the capillary function

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10
Q

Capillary function due to disease

A

Capillaries are fragile in case of VASCULITIS or other diseases that affect the wall of blood vessels

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11
Q

What does bleeding time (BT, BMBTT) depend on?

A

Depends on

Thrombocytic function, the Platelet count, and the Capillary function.

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12
Q

What is Bleeding time a test for

A

1. Thrombocytopenia

2. Thrombocytopathy

3. Vasopathy

NOT COAGULOPATHY

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13
Q

What is normal BT

A

3-5 min

No danger in clinical bleeding if the platelet count is above 50*10^9/L

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14
Q

What is Coagulation time (CT) a test for

A

Test for COAGULOPATHIES

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15
Q

In CT test what blood do we use

A

Fresh, native (not anticoag) whole blood sample

Not cause increased tissue factor (Factor III)

= initiates coagulation cascade

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16
Q

In CT what methods do you use

A

2 syringe method

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17
Q

In CT - where/how can you examin it

A

The appearance of Fibrin strand

CT on watch glass

CT in plastic syringe

CT in glasstube

CT in ACT tube

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18
Q

In CT - when does the first fibrin strand appear

A

Within 1-2 min

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19
Q

In CT - on watch glass

A

Treated with paraffin or wax

(scratch may initiate coagulation cascade)

Solid like gelatin

7-15min

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20
Q

In CT - in plastic syringe - time

A

10-12min

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21
Q

In CT - in glasstube - time

A

4-5 min

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22
Q

In CT - in ACTtube - time

A

3min

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23
Q

CT in ACT (activated clotting time) tube

A

SiO2 in 37 degrees –> Activates Factor XII (Hagemann, contact factor)

Factor XII activates Factor IX –> Kallikreinogen –> Kinigogen

= Fibrolytic Pathway

Move tube every 15-20sec

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24
Q

What is platelet (Thrombocytic) Count important for

A

Especially when BT, BMBTT is INCREASED

Petechia are visuable on skin or mucous membranes

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25
Q

Platelet (Thrombocytic) count is measured from which blood?

A

ANTICOAGULATED BLOOD

Na2+, K2, EDTA

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26
Q

Platelet (Thrombocytic) Count

Methods

A

3 Methods

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27
Q

Platelet (Thrombocytic) Count

Method 1

A
  1. 1 ml EDTA anticoagulated blood sample
  2. 9 ml physiological saline solution,

sedimented for 2 hours.

upper layer

Bürker chamber (hemocytometer).

count the number of platelets in 10 rectangles.

the number should be multiplied by 10^9

= number of platelets in 1 liter blood.

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28
Q

How can you make the process quicker in case of platelet count

A

the process can be made quicker if the sample in NaCl-solution is centrifuged on 1500/min.

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29
Q

Thrombocyte counting in
Bürker chamber is not accurate.

True or False

A

True

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30
Q

How to estimatete Platelet count with

Method 2

A

Platelet count can be estimated by using a blood smear.

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31
Q

Platelet count method 2

Magnification and count

A

Magnification: 1000X

Count: 20*10^9

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32
Q

Platelet count Method 2

This method is also very uncertain, however checking a blood smear for any purpose can be very important.

TRUE OR FALSE

A

TRUE

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33
Q

Platelet count Method 2

What does big thrombocytes tell you

A

The propper fuction

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34
Q

Platelet count Method 2

How is low thrombocytic count measured?

A

Automatic cell counter

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35
Q

Platelet count method 3

How to measure?

A

Platelet count can be measured by using automatic cell counters.

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36
Q

Platelet count Method 3

Which particles are taken as platelets

A

Particles of the blood between 5-30 fl volume
are taken as platelets.

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37
Q

Platelet count method 3

What can happen due to regenerative processes of the bone marrow

(Big platelets)

A

Sometimes in regenerative processes of the bone marrow, when there are many young (big) platelets circulating i.e. in case of

chronic blood loss, or physiologically in cats and in King Charles spaniels

= Average thrombocytic volume can be so high, that these big platelets are taken as red blood cells by the counter.

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38
Q

Platelet count method 3

Why are evaluation of blood smares important

A

an important step of the diagnosis of thrombocytic disorders!

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39
Q

Platelet count

What are the General Platelet Count

A

200-800 x10^9/l

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40
Q

What are the Major causes of thrombocytopenia:

A

1. decreased production of thrombocytes in the bone marrow

2. increased utilization of thrombocytes: DIC (disseminated intravascular coagulopathy)

3. increased destruction of thrombocytes : autoimmune thrombocytopenia (AITP)

4. increased sequestration of thrombocytes: in case of (chronic) splenomegaly

5. increased loss of thrombocytes: subacute/chronic bleeding

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41
Q

Clot Retraction Test

If you leave the blood in a ube for some hours, what will happen?

A

If you leave the blood clot in a tube for some hours, it will become smaller, and serum will appear around the
clot.

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42
Q

Clot retraction test

If you leave the blood clot in a tube for some hours, it will become smaller, and serum will appear around the
clot. But WHY?

A

The reason for this clot retraction is that platelets contain a contractile protein called thrombostenin,

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43
Q

Clot retraction test

What is Normally the volume of serum released by the clot within one hour

A

approx. 25% of the whole volume of the
initial clot..

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44
Q

How can we measure Thrombocytic function

A

Estimated by performing Clot Retraction time test

If the clot retraction is slower or does not
happen at all, we can suspectthrombocytopathy.

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45
Q

When do we use the Platelet aggregation test

A

When we suspect thrombocytopathy, i.e. von Willebrand disease.

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46
Q

Using the Platelet aggregation test what do we use, and to estimate what?

A

use aggregometer to estimate the aggregating ability of platelets correctly.

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47
Q

Platelet aggregation test

What type of blood should you prepare?

Where do you find the platelet rich plasma?

Propperties of the fluid?

A

We have to prepare a citrated blood sample and the
upper layer should be used for this analysis.

This is the platelet rich plasma.

This fluid is slightly opaque.

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48
Q

Platelet aggregation test

Which drugs causes exaggerated aggregation of platelets

A

ADP,
epinephrine etc.

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49
Q

Platelet aggregation test

What can be analysed by a spectrophotometer.

A

The speed and the rate of the clearing can be analysed by a spectrophotometer.

Nummerical value

50
Q

Thrombocytic morphology

Size

A

Thrombocytes have 1-2 μm diameter.

Horses, sheep, cattles, have the smallest platelets (3-5 fl),

Dogs and swine have bigger (7-8fl),

Cats have the biggest (10-15fl) thrombocytes.

51
Q

Thrombocytic morphology

Center

Edges

A

Center= GRANULOMER

Edges= HYALOMER

52
Q

Major causes of thrombocytopathies:

A
  1. improper development of platelets,

for example because of hereditary glycoprotein deficiencies, etc.

  1. von Willebrand’s disease (see later)
  2. Uraemia, liver failure,myelo-, and/orlymphoproliferative diseases,NSAID treatment, etc.
53
Q

In case of thrombocytopenia, thrombocytopathies and vasopathies

  • What can we expect?
A

Besides normal coagulation, we cannot expect
signs of a really severe bleeding disorder

(suffusion, haematoma, haemothrorax, haemoperitoneum),

Because these are prevented by the formation of polymerized fibrin strands (fibrin thrombus) at the end of the coagulation
cascade.

54
Q

In case of COAGULOPATHY

- What can we expect?

A

Besides normal thrombocytic and blood vessel function and normal platelet count may lead
to the aforementioned severe bleedings, and sometimes bleeding to death, because thrombocytic thrombi are not
stable without a fibrin network, and in case of a big blood vessel injury, thepower of blood flow can sweep away
the thrombocyte-thrombus from the wound!

55
Q

Major alterations of the basic tests in different haemostasis disorders:

COAGULOPATHY

A

INCREASED CT

Normal BT, PC (platelet count) and Platelet morphology

56
Q

Major alterations of the basic tests in different haemostasis disorders:

THROMBOCYOPENIA

A

INCREASED BT, DECREASED PC

Normal CT and Platelet morphology altered or not

57
Q

Major alterations of the basic tests in different haemostasis disorders:

THROMBOCYTOPATHY

A

INCREASED BT

Normal CT, PC (platelet count), and Platelet morphology altered

58
Q

Major alterations of the basic tests in different haemostasis disorders:

VASCULOOPATHY

A

INCREASED BT

Normal CT, PC (platelet count), and Platelet morphology

59
Q

How do you examine Coagulopathies

A

By using more specific “global “tests, so that we can evaluate, which group(s) of factors are not functioning properly.

Citrated blood should be used (citrate prevents coagulation by binding
calcium ions), which means, 3,8 % Na-citrate:blood = 1:9. (Prepared tubes are commercially available, usually
blue cap)

60
Q

What is the ratio of a prepared citrate sample?

A

3,8 % Na-citrate:blood = 1:9.

61
Q 
Prothrombin time (PT):
When do you have to perform it
A

This test must be performed within 1 hour after sampling.

62
Q

Prothrombin Time (PT)

Bloodsample mix

A

Blood samples should be mixed with 3.8% sodium
citrate in 9:1 dilution (4.5 ml of blood and 0.5 ml of Na-citrate).

Then centrifuge the samples in 3000 rpm for 10 min. and separate the (decalcinated) plasma from the sediment.

The plasma must be kept on 37 degrees.

63
Q

Prothrombin Time (PT)

What does the reagent contain

A

The reagent (Simplastin) for PT evaluation contains:

rat uterus tissue homogenate as tissue thromboplastin (Factor III.), and CaCl2. The reagent must be kept at 37 degrees before use.

64
Q

Evaluation of Prothrombin time (PT)

A

The evaluation can be performed by using a

1. coagulometer (see on the practical!) or in

2. test-tube.

200 μl reagent
should be mixed with 100 μl decalcinated (citrated!) plasma and the time of coagulation should be noted.

65
Q

Normal Prothrombin time (PT)

A

Normal PT: 10-15 sec.

66
Q

What does the Prothrombin time (PT) give information about

A

The function of the extrinsic pathway,

  • because the coagulation cascade is triggered by adding tissue factor (and calcium ion) to the dacalcinated plasma sample.
67
Q

Factors involved in Prothrombin time (PT) are:

A

VII., X., V., II., I., XIII.

68
Q

Activated partial thromboplastin time (APTT):
For this test decalcinated plasma should be used similarly to PT.

True or false

69
Q

The reagent of Activated partial thromboplastin time test (APTT) contain

A

The reagent (Silimat) contains rabbit brain homogenate as PF3 (platelet factor 3) and

Micronised silica as contact activator.

70
Q

Activated partial thromboplastin time (APTT): The MIX

A

100 μl decalcinated plasma should be mixed with

100 μl reagent and

the mixture kept on 37 degrees

then 100 μl of 0.025 mmol/l CaCl2 solution should be added and from this moment the time of coagulation should be noted.

71
Q

Normal Activated partial thromboplastin time (APTT):

A

Normal APTT: 20-30 sec..

72
Q

What does the Activated partial thromboplastin time (APTT) give us information about?

A

The function of the intrinsic pathway

because the cascade is triggered by providing surface activation (imitating the effect of free collagen, which appears on the inner surface of the vessels in case of blood vessel injury), and adding platelet factor 3 (PF3) and Ca 2+ for the activation of factor X. (remember the
coagulation cascade!!)..

73
Q

Factors involved in Activated partial thromboplastin time (APTT) are?

A

XI., IX., VIII., X., V., II., I., XIII.

74
Q

Thrombin time (TT) depends on

A 
Thrombin time (TT)
By performing this test, **decalcinated plasma should be simply mixed** with a reagent **containing thrombin only**.

In this case coagulation time depends on the concentration of fibrinogen and Factor XIII.in the plasma.

Obviously, this test can also be used for the estimation of fibrinogen concentration (suspecting a normal factor XIII. level in
the blood).

75
Q

Intrinsic pathway problem

A
  1. hemophilia A-factor VIII. deficiency;
  2. hemophilia B-factor IX. deficiency;
  3. von Willebrand’s disease)
76
Q

Intrinsic pathway problem
APTT

PT

A

APTT increased

77
Q

Extrinsic pathway problem

A
  1. Factor VII deficiency, dicumarol toxicosis - first stage)
78
Q

Extrinsic pathway problem
APTT

PT

A

PT INCREASED

79
Q

Common pathway problem

A

  1. Liver disease
  2. decreased. prod. of coag. factors,
  3. DIC, dicumarol toxicosis - second stage,
  4. Factor X. and/or V. and/or II. and/or I. and/or
    * *XIII**. deficiency)
80
Q

Common pathway problem

APTT

PT

A

APTT and PT INCREASED

81
Q

In the early stage of dicumarol (or warfarin) toxicosis both APTT and PT is increased,

True or false

A

False

In the early stage of dicumarol (or warfarin) toxicosis only PT is increased,

82
Q

In the early stage of dicumarol (or warfarin) toxicosis only PT is increased, and later APTT is increased, too.

True or false

83
Q

What is the competitive antagonist of vitamin K.

84
Q

What is VIT D responsible for

A
  • *Gamma-carboxylation** of:
  • *1. Proconvertin** (Factor VII),

2. Christmas (Factor IX),

3. Stuart-Prower (Factor X) and the

4. Prothrombin (Factor II), as
they are Ca2+ dependent factors,

85
Q

Vitamin K deficiency causes ?

A

The inability of these factors to bind calcium.

86
Q

Which factor will be deficient first.

A

Factor VII. has the shortest half life, so this factor will be deficient first.

87
Q

Which test will show the deficiency problem first.

A

When the Prothrombin Time is increased when
there is factor VII deficicency, so this test will show the problem first.

88
Q

What is the Function of Vitamin K

A

To exaggerate the post-synthetic gamma-carboxylation of those factors in the liver.

This makes these factors to be able to bind Ca2+, and thus become
functionally active.

89
Q

In case of vitamin K deficiency of any origin, improperly carboxylised prothrombin can be detected by using ?

A

ELISA (Enzyme Linked Immunosorbent Assay) test, called PIVKA II. (Proteins Induced by Vitamin K Absence)

90
Q

Fibrin degradation products (FDP)
What is Fibrinolytic pathway is responsible for

A

The Fibrinolytic pathway is responsible for keeping the clot formation within normal limits.

91
Q

What are the Clot inhibitors

A

(antithrombin III., alpha2-macroglobulin, alpha1-antitripsin, heparin – the latter increases the binding of antithrombin III to thrombin)

are able to bind to thrombin and neutralize it.

92
Q

After complete coagulation, the clot is usually not needed anymore, so it should be broken down by …….

A

fibrinolytic enzymes.

93
Q

What are the activators of factor XII. (Hageman).

A

1. Free collagen fibres, (exposed at the site of blood vessel injury),

2. kininogen and

3. kallikrein are the activators of factor XII. (Hageman).

94
Q

XII.a (activated form of factor XII. ) further activate

A

the extrinsic pathway,

95
Q

XII.a (activated form of factor XII. ) further activates the extrinsic pathway and is able to form…….

A

kallikrein from prekallikrein.

  • *Kallikrein** activates the kininogen system, also, forming
  • *Bradykinin** (an activated form of kininogen), which is a very potent mediator of pain.
96
Q

What is an activated form of kininogen, and what is it a strong mediator for?

A

Bradykinin (an activated form of kininogen), which is a very potent mediator of pain.

97
Q

What is the most
important activator of plasminogen.

A

Kallikrein

98
Q

What is the activated form of plasminogen?

A
  • *Plasmin** (activated form of plasminogen) is an endopeptidase, which can
  • *cleave fibrin strands into small pieces.**
99
Q

\

Before the total degradation of polymerized fibrin strands, increased
level of

A

fibrinolysis-products, fibrin dimers and monomers (so called: “fibrin degradation products or proteins,
FDP”) can be measured in the blood.

100
Q

What is the fuction of FDP

A

Fibrinolyticenzyme removes clot - elevates level of FDP

but its not accurate because the degradation product could come from both - fibrinogen/fibrin

101
Q

What to use instead of FDP ?

A

D-Dimer level

To distinguish btw fibrin and not fibrinogen

102
Q

What is helpfull with D-Dimer

A

Helpful in early diagnosis of disseminated Intravascular coagulopathy (DIC)

103
Q

What is DIC

A

DIC is a common acute disorder

(that requires accurate and quick laboratory diagnosis)

Usually a secondary disease, caused by primary diseases

LIFETHREATHENING

104
Q

Name some primary causes that could have caused DIC

A

Septicemia

Pancreatitis

Burn injuries

Necrosis of big tumors

Shock

Polytraumatisation

105
Q

What could be the first sign of DIC

A

Positive FDP or D-dimer test

106
Q

What is present simultaneously many places in the body during DIC

A

Formation of Microthrombus + Fibrinolysis

Because of severe damage(necrosis) or Blood vessel injuries

107
Q

What is “consumptional coagulopathy”.

A

In case of DIC, microthrombus formation and fibrinolysis are
present at many different places in the body simultaneously, so coagulation factors and platelets are consumed very quickly during this process.

108
Q

What happens during Consumptional Coagulopathy

A

Coagulating factors and platelets are consumed

109
Q

Lab DIAGNOSIS of DIC

CT, BT, Platelet count, PT, APTT and TT?

FDP and D-Dimer?

SCHYSOCYTES and/Or BURR CELLS

A

INCREASED

CT, BT, PT, APTT and TT

FDP and D-Dimer

DECREASED

Platelet count

SCHYSOCYTES and/Or BURR CELLS

in blood smare = damaged red blood cells

110
Q

Diagnosis of Von WILLEBRAND Disease

Which species

A

Human and dog (Dobberman Pincher)

111
Q

Diagnosis of Von WILLEBRAND Disease

Which factor is deficient

A

Von Willebrand factor

FACTOR VIII is deficient

112
Q

Diagnosis of Von WILLEBRAND Disease

What are the 3 main parts of complete factor VIII

A
  1. Von Willebrand factor – platelet adhesion and aggregation,
  2. VIIIc – is the antihemophilic factor, and the
  3. Factor VIII related antigen – is the hapten that is the determinable
    part, and is bound strongly to von Willebrand factor.
113
Q

How does BT(BMBT), Clot reaction ability look like in the case of a dog with VON WILLEBRAND DISEASE

A

INCREASED

BT, BMBT

DECREASED

Clot reaction ability

Coagulation disorder sometimes

114
Q

The specific diagnosis of VON WILLEBRAND DISEASE is based upon

A

The detection of the lack of von Willebrand-related antigen.

115
Q

What happens to BT, Platelet count, APTT and PT due to

THROMBOCYTOPENIA

A

BT is INCREASED,

Platelet count is DECREASED

APTT and PT = Unchanged

116
Q

What happens to BT, Platelet count, APTT and PT due to

THROMBOCYTOPATHY

A

BT is INCREASED,

Platelet count, APTT and PT is UNCHANGED

117
Q

What happens to BT, Platelet count, APTT and PT due to

  • *Intrinsic pathway disorder**
    (pl. haemophilia A, B)
A

INCREASED APTT

BT, Platelet count and PT is unchanged

118
Q

What happens to BT, Platelet count, APTT and PT due to

Factor VII Deficiency

A

Factor VII Deficiency (in early phase of DICUMAROL toxicosis)

PT is INCREASED

BT, Platelet count and APTT is unchanged

119
Q

What happens to BT, Platelet count, APTT and PT due to

Disorders of the common pathway

A

Disorders of the common pathway

(Liver faiure, Factor X deficiency, late stage of DICUMAROL toxicosis)

BT is unchanged or INCREASED,

APTT and PT is INCREASED

The platelet count is unchanged

120
Q

What happens to BT, Platelet count, APTT and PT due to

DIC

A

BT, APTT and PT is INCREASED

Platelet count is DECREASED

121
Q

What happens to BT, Platelet count, APTT and PT due to

VON WILLEBRAND-DISEASE

A

BT is INCREASED

APTT is unchanged or INCREASED

Platelet count and PT is unchanged

Lab D (11 decks)

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