Professional Documents
Culture Documents
Membranous system
Dense tubular system
Derived from rough ER and sequesters or hold calcium for platelet activation process and prostaglandin
synthesis
Circulating platelets These are platelets found in the circulation and ca be counted.
Circulating, resting platelets are biconvex, although the platelets in blood collected using the
The proplatelet process sheds platelets, cells consisting of granular cytoplasm with a membrane
but no nuclear material, into the venous sinus of the bone marrow.
Thrombocytopoiesis A process where Platelets are released into the bone marrow through shedding
from megakaryocyte proplatelet processes
HEMOSTASIS
Hemostasis
Process that retains the blood within the vascular system during periods of injury, localizes the reaction involved in the
site of injury, and repairs and re-establishes blood flow through the injured vessel.
The maintenance of circulatory hemostasis is achieved through the process of balancing bleeding (hemorrhage) and
clotting (thrombosis).
A. Extravascular Component
- Involves the tissues surrounding a vessel.
- The ability of the surrounding tissues to aid in hemostasis depends on the following factors:
a. Bulk or amount of the surrounding tissue
b. The type of tissue surrounding the injured vessel
c. The tone of surrounding tissue
B. Vascular Component
- Involves the blood vessels in which the blood flows.
a. Capillaries
b. Arteries
c. Veins
C. Intravascular Component
- The key component in intravascular hemostasis are platelets and biochemical (procoagulants) in the plasma.
Hemostasis, the arresting of bleeding, depends on several components. The four major components are the r
vascular system,
- blood coagulation factors, and-
platelets (thrombocytes), r fibrinolysis and ultimate tissue repair. Three other, less important,
components are the complement and kinin systems as well as serine protease inhibitors. (Turgeon)
PRIMARY HEMOSTATIS
-Primary hemostasis is initiated by the exposure of platelets to the subendothelial connective tissue components blood vessels
(Collagen, microfilaments, basement membranes).
VASCULAR RESPONSE
Vascular injury to a large or medium-size artery or vein requires rapid surgical intervention to prevent exsanguination.
When a smaller vessel, such as an arteriole, venule, or capillary, is injured, contraction occurs to control bleeding. This
contraction of the blood vessel wall is called vasoconstriction.
Minimal interactions leading to platelet activation or clot formation occur between the circulating blood and intact
endothelial surfaces. However, disrupted endothelial cells release thromboplastic substances that can initiate coagulation.
Collagen, in particular, initiates contact activation of factor XII, thereby initiating blood coagulation.
Vascular integrity or the resistance to vessel disruption requires three essential factors. These factors are-circulating
functional platelets, -
adrenocorticosteroids, andr ascorbic acid. A Lack these factors produces fragility of the vessels,
which makes them prone to disruption
PLATELET RESPONSE
Platelets have an average diameter of 2 to 4 um, with younger platelets being larger than older ones. In contrast to
megakaryocytes platelets have no nucleus. The cytoplasm is light blue, with evenly dispersed, fine red-purple granules.
An inactive or unstimulated platelet circulates as a thin, smooth-surfaced disc. This discoid shape is maintained by the
microtubular cytoskeleton beneath the cytoplasmic membrane.
Platelets are extremely sensitive cells and may respond to minimal stimulation by forming pseudopods that spontaneously
retract. Stronger stimulation causes platelets to become sticky without losing their discoid shape; however, changes in
shape to an irregular sphere with spiny pseudopods will occur with additional stimulation. This alteration in cellular shape
is triggered by an increase in the level of cytoplasmic calcium.
Agonists that lead to platelet activation are varied and include a- nucleotide (ADP), r
lipids (thromboxane A2, platelet-
activating factor), a
-structural protein (collagen), and r
a proteolytic enzyme (thrombin).
Platelet Adhesion When vascular injury occurs, platelet come in contact with subendothelium (collagen, fibronectin).
vWf binds to glycoprotein Ib/IIb or the GP Ib/IX/V complex on platelet surface
If vascular injury exposes the endothelial surface and underlying collagen, platelets adhere to the
subendothelial collagen fibers, spread pseudopods along the surface, and clump together
(aggregate). Platelet adhesion to subendothelial connective tissues, especially collagen, occurs
within 1 to 2 minutes after a break in the endothelium.
Platelet Activation Morphological and functional change in plates
Cyclooxygenase (from the platelets) metabolizes arachidonic acid to form prostaglandin
enderoperoxides, which are converted to thromboxane A2 (a vasoconstrictor and a platelet
stimulator, causing platelet secretion and aggregation)
Aspirin inhibits cyclooxygenase pathway (remission after 7 to 10days)
Platelet secretion Following activation, platelet undergoes a shape change most probably caused by contraction of
microtubules
From a disk shape to spherical shape with the extrusion of numerous pseudopods
Platelet granules move to the center of the platelets and fuse with the open canalicular system
connected to the outside of the platelet; in this way the content of the granules are extruded to the
outside
Platelet Simultaneously with platelet release, platelet stimulating agents (collagen, ADP, epinephrine,
aggregation thrombin) bind to the platelets, causing then to adherence to one another
Platelet stimulating agents such as collagen, ADP, epinephrine, thrombin binds to platelets, causing
them to adhere to one another. Fibrinogen binds to GP IIb/ IIIa receptors on adjacent platelets and
joins them together in the presence of ionized calcium
Fibrinogen is necessary as cofactor for platelet aggregation
Bridges formed by fibrinogen in the presence of calcium produce a sticky surface on platelets. This
results in aggregation. If these aggregates are reinforced by fibrin, they are referred to as a
thrombus
SECONDARY HEMOSTATIS
COGULATION MECHANISM
Thought of as a complex series cascading reactions involving development of enzymes from their precursor (zymogens,
procoagulants, proenzymes)
Most of the substances necessary for coagulation are present in an inert form and must be converted to an activated
state.
As one enzyme is formed it the becomes available to convert the next zymogen to its activated enzyme (serine protease)
Factors have been given a Roman-numerals. They are numbered in the approximate order of their discovery
When a procoagulant becomes for instance, activated factor VII
is VIIa. Both zymogens and cofactors become activated in the coagulation process
The initiation of the coagulation process may occur via one of two pathways: the extrinsic pathway and the intrinsic
pathway. Regardless of the initiating pathway, the two pathways converge into a final common pathway. The outcome of
this process is the conversion of circulating insoluble coagulation factors into a gelatinous fi brin clot with entrapped blood
cells, a blood clot. As repair of damaged tissue takes place, the clot is lysed and the particulate matter is removed by the
mononuclear phagocytic system.
STAGES
Stage 1 Generation of prothrombinase complex
Stage 2 Conversion of prothrombin to thrombin
Stage 3 Conversion of fibrinogen to fibrin
EXTRINSIC PATHWAY
initiated by the entry of tissue thromboplastin into the circulating blood. Tissue thromboplastin is derived from
phospholipoproteins and organelle membranes from disrupted tissue cells. These membrane lipoproteins, termed tissue
factors, are normally extrinsic to the circulation.
Factor VII binds to these phospholipids in the tissue cell membranes and is activated to factor VIIa, a potent enzyme
capable of activating factor X to Xa in the presence of ionized calcium.
FUNCTIONS OF THROMBIN
Procoagulant induces platelet activation and aggregation
activates cofactor VIII to VIIIa
activates factor XIII to XIIIa
converts prothrombin to thrombin via autocatalysis
Coagulation Inhibitor The coagulation inhibition activity displayed by thrombin is the binding of AT-III to inhibit
serine proteases and binding to thrombomodulin to activate protein C.
Promotion of endothelial cell release of t-PA (Tissue plasminogen activator).
Tissue repair Thrombin mediates tissue repair by inducing cellular chemotaxis and stimulation of
proliferation of smooth muscle and endothelial cells.
Factor VIII can be subdivided into various functional components. The total molecule, consisting of both a high molecular-
weight fraction and a low molecular-weight fraction, is described by the nomenclature VIII/vWF. Factor VIII/vWF consists of
two major moieties. The high molecular-weight moiety consists of the vWF, VIIIR:RCo, and VIIIR:Ag components. The low
molecular-weight moiety consists of the VIII:C and VIIIC:Ag components.
Factor VIII:C has procoagulant activity as measured by clotting assay techniques.
Factor VIII/vWF multimers form ionic bonds with factor VIII:C and transport VIII:C in the circulation.
Factor VIIIC:Ag is a procoagulant antigen as measured by immunological techniques using antibodies for factor VIII:C.
Factor VIIIR:Ag is a related factor VIII antigen that has been identifi ed using immunological techniques employing
heterologous antibodies to VIII/vWF.
Factor VIIIR:RCo demonstrates ristocetin cofactor activity, which is required for the aggregation of human platelets
induced by the antibiotic ristocetin
ADDENDUM
Factor IX is a stable protein factor that is neither consumed during clotting nor destroyed by aging at 4°C for 2 weeks
Factor XIII, an acute-phase reactant, is consumed during the clotting process and is not found in serum. Factor VIII is
extremely labile, with a 50% loss within 12 hours at 4°C in vitro and a similar 50% loss in vivo within 8 to 12 hours after
transfusion. In addition, factor VIII can be falsely decreased in the presence of lupus anticoagulant (LA).
Factor V is an extremely labile globulin protein. It deteriorates rapidly, having a half-life of 16 hours
A large quantity of thrombin is consumed during the process of converting fibrinogen to fibrin. A unit of thrombin will coagulate
1 mL of a standard fibrinogen solution in 15 seconds at 28°C.
Tissue thromboplastin is the term given to any non-plasma substance containing lipoprotein complex from tissues.
These tissues can be from the brain, lung, vascular endothelium, liver, placenta, or kidneys; these tissue types are capable
of converting prothrombin to thrombin
Factor VII, a beta-globulin, is not an essential component of the intrinsic thromboplastin-generating mechanism. It is not
destroyed or consumed in clotting and is found in both plasma and serum, even in serum left at room temperature for up to 3
days. The action of factor VII is the activation of tissue thromboplastin and the acceleration of the production of thrombin from
prothrombin. This factor is reduced by vitamin K antagonists
intrinsic
APTT= assess + common
pathway
Reptilase and Thrombin time (TCT)= assess
fibrinogen deficiency
Stypven time= for common
pathway
EXAMPLE CASES:
a. Fibrinogen deficiency
b. Factor IX deficiency
c. Factor VII deficiency
d. Factor XIII deficiency
MIXING STUDIES
a. Lupus anticoagulant / circulating anticoagulant
b. Fibrinogen deficiency
c. Factor II deficiency
d. Factor VII deficiency
D 1. PT Abnormal, PTT normal, Adsorbed plasma not corrected, Aged serum corrected, Fresh plasma corrected=
C 2. PT Abnormal, PTT Abnormal, Adsorbed plasma not corrected, Aged serum not corrected, Fresh plasma corrected =
B 3. PT Abnormal, PTT Abnormal, Adsorbed plasma corrected, Fresh serum not corrected, fresh plasma corrected =
A
4. PT Abnormal, PTT Abnormal, Fresh plasma not corrected, Fresh serum not corrected, Adsorbed plasma not corrected =
3. Common Pathway ( 1,11 V X )
( V" ' " ) , ,
2. Common Pathway ( V ,
X , 1,11 )
Adsorbed =
10,9 7,2 ,
forum 1 2,5, 8 13
Aged = , ,
K+
1- Caz-1
Factor I, V, VIII, XIII Factor VII, IX, X, II Factor XII, XI, HMKW, PK
Consumed during coagulation Requires vitamin K Not consumed during
Absent in serum Not consumed during coagulation
Not absorbed by barium sulfate coagulation Not absorbed by barium sulfate
or aluminum hydroxide Absorbed by barium sulfate or or aluminum hydroxide
Increases in inflammation, aluminum hydroxide
pregnancy, stress and fear, oral Increases in pregnancy, and
contraceptives oral contraceptives
NOTE Vitamin K catalyzes an essential posttranslational modification of the prothrombin group proteins:
gamma-carboxylation of amino-terminal glutamic acids
FIBRINOLYSIS
Primary purpose is to digest fibrin clots as they are formed in order to keep vascular system free of deposited fibrin and
fibrin clots
Occurs when plasminogen is converted to plasmin, which dissolves the fibrin or fibrinogen into smaller fragments termed
FDP (Fibrin degradation product) or FSP (Fibrin split product)
Plasmin a serine protease that systematically digest fibrin polymer by the hydrolysis of arginine-related and lysine
related peptide bonds
Free plasmin- capable of digesting plasma fibrinogen, factor V, Factor VIII, and fibronectin.
Plasminogen is a 92,000 Dalton plasma zymogen produced by the liver. It is a single-chain protein possessing five
glycosylated loops termed kringles. Kringles enable plasminogen, along with activators TPA and UPA, to bind the lysine
moieties on the fibrin molecule during the polymerization process. This fibrin-binding step is essential to fibrinolysis.
Fibrin-bound plasminogen becomes converted into a two-chain active plasmin molecule when cleaved between
arginine at position 561 and valine at position 562 by neighboring fibrin-bound TPA or UPA.
k
I.K Aytona Page 14
SPECIMEN COLLECTION AND PROCESSING FOR HEMOSTATIS TESTING
Methods of collection
Two syringe method
Evacuated tube technique
These two methods are preferred because it requires that a few volume of blood should be discarded first then the next
volume of blood is collected, this is done to prevent possible contamination of tissue fluid that contains thromboplastin
which somehow alters results obtain from clotting test
cyclooxygenase pathway
o Platelet function test: 1week abstinence from the following drugs
o Bleeding time test: 24 hours aspirin intake is prohibited
PRP Can be obtained by centrifuging WB collected from 3.2% Sodium Citrate at 50g x for 30
(Platelet Rich Plasma) minutes
It contains 200,000 to 300,000 /ul platelets (200 to 300 x109/L platelets)
It is used for platelet aggregometry or platelet function test
PPP Can be obtained by centrifuging WB collected from 3.2% Sodium Citrate at 1500g x for 15
(Platelet Poor Plasma) minutes
It contains less than 10,000/ul platelets
It is used for clot-based coagulation test (E.g PT and APTT)
1. Make a 1:100 dilution by placing 20 mL of well-mixed blood into 1980 ul of 1% ammonium oxalate in a small test tube.
2. Mix the dilution thoroughly and charge the chamber. (Note: A special thin, flat-bottomed counting chamber is used for phase-
microscopy platelet counts.)
3. Place the charged hemacytometer in a moist chamber for 15 minutes to allow the platelets to settle.
4. Platelets are counted using the 40x objective lens (400x total magnification). The platelets have a diameter of 2 to 4 um and
appear round or oval, displaying a light purple sheen when phase-contrast microscopy is used. The shape and color help
distinguish the platelets from highly refractile
background.
5.Count the number of platelets in the 25 small squares in the center square of the grid. The area of this center square is 1
mm2. Platelets should be counted on each side of the hemacytometer, and the difference between the totals should be less
than 10%.
6.Calculate the platelet count
7.The accuracy of the manual platelet count should be verified by performing a platelet estimate on a wright-stained peripheral
blood.
NOTE
A. If fewer than 50 platelets counted on each side = repeat procedure and dilute blood to 1:20
B. If more than 500platelets counted on each side = repeat procedure and dilute blood to 1:200
B. Indirect (Smear)
o Dameshek
o
o
NOTE
1 platelet per 10-40 RBCs
3-10 platelets per 100 RBCs (or in 1 OIF)
5-20 platelets per 200 RBCs
A normal blood smear should demonstrate approximately 8 to 20 platelets per field
b. STEFANINI
o 3-5 ml blood (37 oC)
o 1/2/16/18/24 hours
o NV: appreciable within 1 hour, complete within 18 to 24 hours
c. MacFarlane
o 5 mL of blood, 370C (1hour)
o NV= 44-67%
o CRT = Volume of serum x 100
TV
Inflate BP cuff to a point hallway between the systolic and diastolic pressure (never exceed 100mmHg), maintain pressure
for 5 minutes
Remove BP cuff and wait for 5 to 10 minutes before proceeding, count petechiae
Positive test is found on thrombocytopenia, decreased fibrinogen and in vascular purpura
Whole blood/ Lee and White method (Tilt tube method) N.V = 5-15 minutes
-uses glass tube 13x100mm in size
2.PROTHROMBIN TIME
NV 10-12 Seconds / 12.6 to 14.6 seconds (Rodak )
Determine extrinsic and common pathway factor deficient
PPP + PT reagent
PT reagent: tissue thromboplastin + CaCl2 (calcium chloride)
Begin the time for clot formation after the addition of calcium chloride
-PT reagents, often called thromboplastin or tissue thromboplastin, are prepared from recombinant or affinity-purified tissue
factor suspended in phospholipids mixed with a buffered 0.025 M solution of calcium chloride. A few less responsive
thromboplastins are organic extracts of emulsified rabbit brain or lung suspended in calcium chloride.
-Recently, however, newer generation thromboplastin reagents have been developed that are based on purified,
recombinant human tissue factor that has been reconstituted into phospholipid vesicles
INR CONDITIONS
2.0-2.5 DVT, high risk surgery
2.0-3.0 Hip surgery, femur fracture
2.0-3.0 DVT, Pulmonary embolism, transient ischemic attack
2.5-3.5 Mechanical/ prosthetic heart valves
2.0-4.5 Recurrent DVT and pulmonary embolism, myocardial infarction, arterial disease
The ISI is a calibration parameter that defines the responsiveness of the reagent relative to a World Health Organization (WHO)
International Reference Preparation, which by definition has an ISI of 1.0.
Begin the time for clot formation after the addition of calcium chloride
If the PTT is performed manually, the test should be done in duplicate, and the two results must match within
10%.
It is standard method for monitoring unfractionated heparin therapy
The test is affected and will give a prolong results in the presence of specific inhibitors/Lupus anticoagulant, Fibrin
degradation products or paraproteinemia such as Multiple myeloma
The PTT reagent contains phospholipid (previously called partial thromboplastin) and a negatively charged particulate activator
such as silica, kaolin, ellagic acid, or celite in suspension. The phospholipid mixture, which was historically extracted from rabbit
brain, is now produced synthetically
Prolonged in the presence of FDP/FSP, Heparin, paraproteins and of thrombolytic agent such as streptokinase
Prolonged when fibrinogen is below 100 mg/dl, and when the function of fibrinogen is impaired.
Hypofibrinogenemia, Afibrinogenemia (absence of fibrinogen) and dysfibrinogenemia (presence of fibrinogen that is
biochemically abnormal and nonfunctional) also cause a prolonged TCT
6.REPTILASE TEST
Reference value: 10-15seconds
Reptilase is an enzyme found in the venom of Bothrops athrox snake, capable of converting fibrinogen to fibrin
Test for fibrinogen deficiency/ fibrinogen abnormality
PPP + ATROXIN (reptilase)
Principle: reptilase catalyzes the conversion of fibrinogen to fibrin. In contrast to thrombin, this enzyme cleaves only
fibrinopeptide A.
Prolonged in the presence of FDP/FSP, Streptokinase, and paraproteins
It is NOT AFFECTED by Heparin / Heparin therapy
The reagent is reconstituted with distilled water and is stable for 1 month when stored at 1° C to 6° C
7
Screening test for factor XIII deficiency
Alternative reagents used can be:
1 % monochloroacetic acid
2 % acetic acid
PPP + 5M urea
Normal clot is insoluble to urea for 24 hours
Factor XIII deficiency the clot is dissolved in less than 24 hours
BASIC TERMINOLOGY
Petechiae Purplish red pinpoint hemorrhagic spots in the skin caused by loss of capillary ability to withstand
normal blood pressure and trauma.
Size: 1 mm or less than 3mm
Purpura Hemorrhage of blood into small areas of the skin, mucous membranes, and other tissue.
Size:3mm to 1cm
Ecchymosis Form of purpura in which blood escapes into large areas of skin and mucous membranes, but not into
deep tissues.
It is caused by leakage of a small amount of blood in the tissue around the puncture site
Size: more than 1cm
Epistaxis Nosebleed that are recurrent, bleed from both nostrils, last longer than 10 minutes, or require physical
intervention
Hemarthrosis Leakage of blood into joint cavities
Hematemesis Vomiting of blood
Hematoma Swelling of tumor in the tissue or a body cavity that contains clotted blood.
Results when leakage of a large amount of blood around the puncture site causes the area to
rapidly swell.
Hematuria RBC in urine
Hemoglobinuria Hemoglobin in urine
Melena Stool containing dark red or black blood
Menorrhagia Excessive menstrual bleeding
COAGULATION DISORDERS
A.CLOTTING FACTOR DEFICIENCIES
FACTOR INHERITED COAGULOPATHIES ACQUIRED
COAGULOPATHY
INHERITANCE COAGULOPATHY
PATTERN
I Autosomal recessive Afibrinogenemia Severe liver disease
DIC
Autosomal dominant Dysfibrinogenemia Fibrinolysis
II Autosomal recessive Prothrombin deficiency Liver disease
Vitamin K deficiency
Anticoagulant therapy
V Autosomal recessive Factor V deficiency Severe liver disease
/ Parahemophilia) DIC
Fibrinolysis
VII Autosomal recessive Factor VII deficiency Liver disease
Vitamin K deficiency
Anticoagulant therapy
VIII X-linked recessive Hemophilia A DIC
autosomal dominant Fibrinolysis
IX X-linked recessive Hemophilia B (Christmas Disease) Liver disease
Vitamin K deficiency
Bleeding associated with Primary Hemostasis defect Petechia, purpura, Ecchymoses, Epistaxis (nosebleed), Gingivail/gums
bleeding, Hematemesis (vomiting of blood), Menorrhagia
Bleeding associated with Secondary Hemostasis defect Hemarthroses, Intracranial hemorrhage, hematomas, Post-
(Clotting factor def.) surgical bleeding
Mucocutaneous (systemic) hemorrhage Associated with thrombocytopenia (platelet count lower than 150,000/ul),
qualitative platelet disorders, or vascular disorders
Anatomic (soft tissue) hemorrhage Seen in acquired or congenital defects in secondary hemostasis, or plasma
coagulation factor deficiencies (coagulopathies).
Most common congenital deficiencies VWD, factor VIII and IX deficiencies (hemophilia A and B), and platelet function disorders
Rare congenital deficiencies Inherited deficiencies of fibrinogen, prothrombin, and factors V, VII, X, XI, and XIII
Two group:
a. The specific inhibitors against specific coagulation factors and
b. Non-specific inhibitors such as the LUPUS INHIBITOR anti phospholipid antibody
Tests:
a. Platelet neutralization test
b. Dilute Russel viper venom test (DRVVT)
c. silica-based partial thromboplastin time (PTT),
IN VIVO, THEY ARE ASSOCIATED WITH ARTERIAL AND VENOUS THROMBOSIS/ THEY ARE PROTHROMBIC.
IN VITRO, THEY ARE ANTICOAGULANT.
C.LIVER DISEASE
The bleeding associated with liver disease may be localized or generalized, mucocutaneous or anatomic.
Associated with multiple factor deficiency without the presence of D-dimer
Liver is one of the major sites for coagulation factor synthesis
Liver disease predominantly alters the production of the vitamin K dependent factors II (prothrombin), VII, IX, and X and
control proteins C, S, and Z. Vitamin K catalyzes an essential posttranslational modification of the prothrombin group
proteins: gamma-carboxylation of amino-terminal glutamic acids. In liver disease, these seven factors are produced in
their des-y-carboxyl forms, which cannot participate in coagulation
Platelet count is decreased (<150,000cells/ul) due to sequestration and shortened platelet survival associated with portal
hypertension and resultant hepatosplenomegaly
Platelet aggregation and secretion is affected and suppressed
Factor VII is the first coagulation factor to exhibit decreased activity in liver disease
Factor V is more specific marker of liver disease because it is not affected by Vitamin K deficiency
Prothrombin time (PT) Serves as a sensitive early marker for mild liver disease that can detect factor VII
deficiency and Vitamin K deficiency
The factor V activity assay, performed in conjunction with the factor VII assay, may be used to distinguish liver disease
from vitamin K deficiency
The PT, PTT, thrombin time, fibrinogen concentration, platelet count, and D-dimer concentration are used to characterize
the hemostatic abnormalities in liver disease.
For hemostatic treatment to resolve liver disease, If the fibrinogen level is less than 50 mg/dL, spontaneous bleeding is
imminent, and cryoprecipitate or fibrinogen concentrate may be selected for therapy
Factor V and VII assays may be used in combination to differentiate liver disease from vitamin K deficiency
Normal Factor V and Decreased Factor VII Dietary Vitamin K deficiency
Decreased Factor V and Decreased Factor VII Liver disease
Acute seen in association with obstetric emergencies, intravascular hemolysis, septicemia, viremia, burns, acute inflammation,
DIC crush injuries, dissecting aortic aneurysms, and cardiac disorders
Chronic associated with vascular tumors, tissue necrosis, liver disease, renal disease, chronic inflammation, use of prosthetic
DIC devices, and adenocarcinoma
1. Clotted: will cause falsely shortened clotting times because of premature activation of coagulation factors and
platelets that generate FVIIa and thrombin
2. Lipemia: may cause falsely prolonged clotting times on OD instruments because of interference with light
transmittance
3. Hemolysis: may cause falsely shortened clotting times because of premature activation of coagulation factors and
platelets that generate Factor VIIa and thrombin
4. Icterus (bilirubinemia): indicates liver dysfunction that may lead to prolonged clotting times because of inadequate
clotting factor production; also may interfere with OD instruments
5. Abnormal clot formation: may lead to falsely elevated clotting times because of instrument inability to detect an end-
point
6. No end-point detected: indicates that the instrument was unable to detect clot formation; the specimen may need to
be tested using an alternate methodology
ANTICOAGULANT THERAPY
1. Heparin therapy
Used to prevent of thrombi in veins or to prevent propagation of previously formed thrombi in veins and arteries
Continuous IV infusion has now become the most popular method of administration
ACTION: inhibit thrombin
MONITORED BY APTT or ACT (Activated clotting time)
NEUTRALIZED WITH: ___________________
In recent years, the new oral anticoagulants (NOAs) factor Xa inhibitors apixaban, thrombin inhibitor abigatran, an factor Xa
inhibitor rivaroxaban and factor Xa inhibitor edoxaban have revolutionized anticoagulation therapy.
3. OTHERS
Examples are Hiruden, Lepirudin (Refludan), Argatroban, Danaparoid (Orgaran), Fondaparinux (Arixtra)
Lepirudin levels can be monitored by the Aptt, ECARIN clotting time, or a chromogenic assay based on the
inhibition of thrombin. The Aptt is the most widely used method of monitoring patients. The target range for
anticoagulation is 1.5 to 2.5 times the baseline Aptt
Argatroban binds to thrombin directly and acts as an anticoagulant by blocking the active site on the thrombin
molecule. This drug is monitored by the Aptt. The therapeutic level is 1.5 to 3.0 greater than the baseline Aptt.
Danaparoid is a mixture of heparinoids which only accelerates the binding of factor Xa to antithrombin and
possesses no anticoagulant activity of its own. It is monitored by chromogenic assay.
Fondaparinux is a synthetic pentasaccharide that accelerates the binding of antithrombin to activated factor Xa.
It has no antithrombin activity. Although this drug usually does not require monitoring, it is recommended that it
be assayed by a system based on the inhibition of factor Xa, if monitoring is needed.
ADDENDUM
Kinin system and complement Kinin system is activated by both coagulation and fibrinolytic system
system Both are associated with hemostasis
Pregnancy associated Normal pregnancy beginning at the time of conception is associated with increased
thrombosis concentrations of coagulation factors VII, VIII, and X and von Willebrand factor
Bethesda assay A test for factor VIII inhibitor
Fibrinogen levels Normal concentration = 200 to 400 mg/dl
Normal titer: 1:128 to 1:256
Abnormal titer: less than 1:64
Hemophilia A carrier detection Approximately 90% of female carriers of hemophilia A are detected by measuring the ratio of
FVIII activity to VWF:Ag concentration (FVIII to VWF:Ag).
20% Platelets transport about _______ of circulating factor V, thus the platelet function in factor V
deficiency may be diminished, which is reflected in a prolonged bleeding time test but normal
platelet aggregation.
Factor XIII A clotting factor that is a tetramer of paired a and b monomers
Confirm factor XIII deficiency To confirm factor XIII deficiency, factor activity may be measured accurately using a
chromogenic substrate assay such as the Behrichrom FXIII assay (Behring)
ARTERIAL Thrombosis Lipoprotein a, CRP, Fibrinogen assay, plasma Homocysteine level, Factor VIII levels
predictors
Clotting factors that are APR Fibrinogen and Factor VIII
Deep vein thrombosis The cardinal symptoms for DVT are edema, erythema, pain, and the sensation of heat.
Depending on clot location, symptoms may involve the entire leg, calf, or ankle and foot. Often
the
persist and grow over several days.
Reporting of plasma D-Dimer The plasma D-dimer assay is reported in D-dimer units (DDUs) or fibrinogen equivalent units
assay (FEUs), depending on the nature of the monoclonal antibody (MAB) employed in the selected kit.