Hemostasis Flashcards
What is involved in primary hemostasis?
Platelets
Vessel
What is involved in secondary hemostasis or coagulation?
Coagulation factors
Vessel
What is involved in tertiary hemostasis or fibrinolysis?
Fibrinolytic molecules
Vessel
What are platelets?
Small fragments of megakaryocyte cytoplasm
Describe the composition of platelets
Phospholipid membrane with glycoproteins
Open canalicular system of membrane invaginations
Cytoskeleton is a peripheral microtubule ring and is made of actin and mysoin
Tubular system of ER
α-granules
Dense granules
Glycogen and mitochondria
What is the function of the platelet membrane?
Coagulation and cell to cell interaction
What is the function of the open canalicular system of membrane invaginations in platelets?
Substance trafficking between platelet and plasma
Increase platelet surface
What does the tubular system of ER in platelets do?
Stores Ca for platelet activation and thromboxane production
What do α-granules play a role in?
Hemostasis
Angiogenesis
What are the dense granules in platelets?
Ca Mg ADP ATP Serotonin
What do glycogen and mitochondria do for platelets?
Energy
What plays a role in megakaryopoiesis and thrombopoiesis?
Thrombopoietin and other cytokines
Where do megakaryopoiesis and thrombopoiesis occur?
Bone marrow
Spleen
Lung
What happens with megakaryopoiesis and thrombopoiesis?
Platelets are released directly into the blood
Where is thrombopoietin made?
Hepatocytes (dog)
Renal tubular epithelium
BM stromal cells
What is TPO expressed?
Constitutively
What is the plasma concentration of TPO dependent on?
MPL receptor numbers present in platelets and megakaryocytes
How does inflammation/IL-6 impact TPO?
IL-6 leads to TPO expression in the BM
What are reticulated platelets?
Young platelets
Increased RNA
Less than 24 hours in dogs
What does the concentration of platelets depend on?
Production Consumption Destruction Shift/sequestration Dilution
What are the methods of evaluating platelets?
Blood smear
Blood sample
Platelet count
Mean platelet volume
Describe a blood smear for platelet evaluation
Platelet estimation
Morphologic abnormalities
Describe a blood sample for platelet evaluation
EDTA (not heparin)
Citrate, theophylline, dipyridamole, and adenosine (CTAD tubes) may be useful for cats
8 hours at RT and 48 hours at 4 deg C
Check for platelet clumping
What can be used for platelet count for platelet evaluation?
Impedance counters
Optical or layer flow cytometers
QBC
Hemocytometer
Describe mean platelet volume for platelet evaluation
Varies between analyzer, anticoagulant, and storage time
Presence of cell fragments and large platelets
What is the concentration of platelets like when there is thrombocytopenia?
Concentration is lower than the LRL
What breeds a predisposed to thrombocytopenia?
Greyhounds
Shiba Inus
What does thrombocytopenia reflect?
A pathologic process, it is not a diagnosis
What is present if the thrombocytes are less than 30,000/μL?
Petechiae and ecchymosis Mucosal bleeding (epistaxis, hematochezia, melena), hematuria, and hyphema
What are causes of pseudo-thrombocytopenia?
Not all of the platelets were counted
In vitro platelet activation or aggregation
Presence of large platelets in impedance counters
Cold agglutinins, or anticoagulant-induce, antibody mediated agglutination
What is a thrombocytopenia shift/sequestration?
Reversibly distributed platelets in vascular systems (spleen)
What does thrombocytopenia shift/sequestration result in?
Mild or moderate thrombocytopenia
Splenomegaly
Severe hypothermia
Endotoxemia
What are causes of decreased production of platelets?
Generalized BM or MK-specific process Idiopathic thrombocytopenia of CKCS Drugs (predictable and dose-dependent; idiosyncratic) Infectious Marrow replacement (myelophthisis)
What infections caused decreased production of platelets?
Direct infection of MK (BVD and canine distemper)
Myelosuppressive cytokinesm (EIA)
Unclear (parvo, canine monocytic ehrlichiosis)
FeLV
What kinds of marrow replacement cause decreased production of platelets?
Bone marrow neoplasia
Myelonecrosis
What is the cause of platelet destruction?
Immune-mediated thrombocytopenia (IMT)
Drug induced IMT
IMT associated to infection
Neonatal alloimmune thrombocytopenia
What is found with IMT?
Platelet surface-associated immunoglobulin
What happens to platelets with IMT?
Destruction by MPS
What does IMT result from?
Defective immune system
Defective platelets
Describe idiopathic or primary IMT
Not associated to a detected disease
It could be autoimmune
Common in dogs
Ab may target MKs
What is the criteria for diagnosis of drug induced IMT?
PSAIg present
Plasma Ab bind in vitro to platelets only in presence of the drug
Resolution after drug is discontinued
Relapse after drug is reintroduced
What are drugs that can cause drug induced IMT?
Gold salts, sulfonamides (dogs)
Methimazole and proplyrhiouracil (cats)
Penicillin and TMS (horses)
What are infections that can cause IMT?
Acute canine ehrlichiosis Other Ehrlichia and Anaplasma Rocky Mountain Spotted Fever Histoplasmosis, Leishmaniasis Distemper or modified virus vaccination Equine infectious anemia Babesiosis
What must neonatal alloimmune thrombocytopenia be differentiated from?
Other causes especially sepsis
Describe IMT caused by neoplasia
May include immunologic mechanisms
Lymphoma
Descibe IMT caused by systemic immune mediated disease (i.e. SLE)
SLE: evidence of increase PSAIg
Evan’s syndrome: IMHA with IMT described in dogs
What can cause thrombocytopenia dilution?
Blood loss (acute and severe) Hemodilution
Describe thrombocytopenia associated with blood loss
Thrombocytopenia could be due to consumption
Mild to moderate and self-limiting
What are causes of thrombocytopenia via consumption?
Platelet activation with accelerated consumption or use
Disseminated intravascaulr coagulation (DIC)
Drugs or foreign material
Evenomation
Vasculitis or endocarditis
What results from platelet activation with accelerated consumption or use?
Localized intravascular coagulation
What is a potent platelet activator?
Thrombin
Describe idiopathic thrombocytopenia of CKCS
Congenital
Common in the breed
Macrothrombocytopenia
No clinical bleeding problem
What is thrombocytopenia do to infectious causes frequently associated to?
Endotoxemia
What are bacteria that can cause thrombocytopenia?
Ehrlichia and Anaplasma spp
What is a fungi that can cause thrombocytopenia?
Histoplasma spp
What are protozoa that can cause thrombocytopenia?
Leishmania, Babesia, and Theileria spp.
How do infectious causes result in decreased platelet production?
Direct infection
Immune suppression
BM inflammation
How does Anaplasma platys cause thrombocytopenia?
Infect platelets
Clinical or subclinical
Mild parasitemia
PCR diagnosis
What neoplasma can cause thrombocytopenia?
Carcinomas
Sarcomas
Lymphomas
Leukemias
How do neoplasms cause decreased production of platelets?
Myelophthisis
Myelodysplasia
Estrogen secretion
Chemotherapy
How do neoplasms cause consumtpion of platelets?
DIC
Vasculitis or thrombosis
How do neoplasms cause destruction of platelets?
Secondary IMT
Hemorrhage, sepsis
How do neoplasms cause sequestration of platelets?
Splenomegaly or heptomegaly
Organ congestion
How does hypophosphatemia cause thrombocytopenia?
Hyperalimentation
Decreased platelet survial
Decreased platetlet ATP
How does anaphylaxis cause thrombocytopenia?
Mechanism incompletely characterized
Inflammatory mediators
DIC
Immune-complex interaction with platelets
What is thrombocytosis?
Platelet concentration greater than URL
What can cause thrombocytosis?
Redistribution or increased production (hemic neoplasias)
What is the cause of physiologic thrombocytosis?
Redistribution
What causes an increased production of platelets?
Inflammation (IL-6 induced TPO production) Nonhemic malignant neoplasia Iron deficiency Vinca alkaloids Rebound (recovery from thrombocytopenia) Postsplenectomy Blood loss Hypercortisolemia
What are the functions of platelets?
Primary hemostatic plug Adhesion Aggregation Degranulation Facilitation of coagulation Clot retratction
How do platelets cause adhesion?
Perturbed endothelium or exposed subendothelium
Via vWF binding to GPIb
What are the agonists that cause aggregation with platelets?
ADP
Collagen
PAF
Thrombin
What are the high affinity sites for cagulation?
Enzymes
Cofactors
Zymogen
What is involved with the contractile process of clot retraction?
Activated platelets
GPαIIbβ3 via fibrinogen
Actin and myosin
What is the standardized primary hemostasis test?
BMBT
What is the BMBT procedure?
Small cut that is 5 mm x 1 mm of the upper lip
Start timing when the cut is made (1-5 minutes)
Remove excess blood with a filter paper without disturbing the cut
The end point is when no blood is transferred to the filter paper
What can BMBT detect?
Only moderate to marked primary hemostasis defects
What could it mean if BMBT is prolonged (>4-5 minutes)?
Thrombocytopenia Thrombocytopathia (platelet dysfunction) vWD Anemia Vascular disease Antiplatelet drugs Afibrinogenemia
What is von Willebrand Factor responsible for?
Platelet adhesion
Platelet aggregation
What is vWF produced by?
Endothelial cells
What does vWF from with Factor VIII?
Noncovalant complexes
What is the purpose of the noncovalent complexes formed by vWF and factor VIII?
It is protective and a stabilizer for factor VIII
What is the most common hereditary bleeding disorder of dogs?
von Willebrand disease
What is type 1 vWD?
All vWF multimers are present in decreased concentration
Dioberman Pinchers and many other breeds
What is type 2 vWD?
Deficiency of vWF with disproportionate decrease of large multimers
German Shorthaired Pointers, German Wiredhaired Pointers, and horses
What is type 3 vWD?
Absence of all multimers
Chesapeaks Bay Retrievers, Dutch Kooikers, Scottish Terriers, Shetland Sheepdogs
What are the clinical and lab findings of vWD?
Mild to severe mucosal bleeding Hemarthrosis and hematomas in horses Absence of petechiae Prolonged BMBT Possibly decreased PTT due to decrease of factor VIII
What should be used to analyze vWF?
Sodium citrate or EDTA
How long is the vWF antigen stable at room temperature?
8 hours
What is coagulation?
Interconnected series of reactions that forms thrombin, which converts soluble fibrinogen into soluble fibrin
What are the enzymatic factors involved in coagulation produced by?
Hepatocytes
What enzymatic facotrs are vitamin k dependent?
2, 7, 9, and 10
Which enzymatic factor is sex-linked (x chromosome)?
Factor 9
What are the nonenzymatic factors involved in coagulation?
Tissue factor
Fibrinogen
FV and FVIII (cofactors)
What is tissue factor?
Cofactor for FVII
Major activator of coagulation in vivo
What is fibrinogen?
Positive acute phase protein produced by hepatocytes
What are FV and FVIII (cofactors)?
Markedly accelerate coagulation
Facilitates surface attachment of other factors
Which nonenzymatic factor is sex-linked?
FVIII
When are FV and FVIII consumed?
During coagulation
What is the extrinsic pathway initiated by?
Tissue factor that activated factor VII
What does the extrinsic pathway initiate?
Common pathway (factor X) Intrinsic pathway (factor 9)
What is the extrinisc pathway important in?
Initiating thrombin generation
What is the intrinsic pathway initiated by?
HMWK, PK, and factor 12 contacting negatively charged surfaces
What does the intrinsic pathway start?
Common pathway by activating factor 10
What is the intrinsic pathway inhibited by?
ATIII (2, 9, 10, 11)
What is the intrinsic pathway activated by?
Factor 7a, which activates factor 9
What does thrombin activate in the intrinsic pathway?
Factors 5, 8, and 11
What is the common pathway initiated by?
Activation of factor 10
What does the common pathway lead to?
Formation of thrombin from prothrombin
What does thrombin do in the common pathway?
Forms fibrin monomers and them multimers cross linked by factor XIIIA
What is the common pathway inhibited by?
AT3
What is antithrombin 3 produced by?
Hepatocytes
What does AT3 do?
Binds, inactivates and removes coagulation factors (most importantly 2a, 9a, and 10a)
What is AT3 markedly enhanced by?
Heparin
What kind of sample is needed to determine the ACT (activated coagulation/clotting time)?
Whole blood
What is ACT?
Screening coagulation test that evaluates surface activation (intrinsic and common)
What can increase the ACT?
Thrombocytopenia
What can ACT be used to monitor?
Heparin therapy
What does the activated partial thromboplastic time (aPTT) screen for?
Intirinsic and common pathways
What is the sample needed for aPTT?
Citrated blood (plasma)
What does prothrombin time screen for?
Extrinsic and common pathways
What is the samples needed for prothrombin time?
Citrated blood (Plasma)
What does PT test?
Ca, phospholipids, and TF
What does thrombin time asses?
Transformation of fibrinogen in fibrin without the interference of thrombin generation
What is the sample neededfor thrombin time?
Citrated blood (plasma)
What is the test done for TT?
Thrombin is added to the sample
What does it mean if there is prolonged time with a TT?
Hypofibrinogenemia or afibrinogenemia Increased consumption, localized coagulation, or DIC Hereditary deficiency Dysfibrinogenemia Heparinized sample Increased FDPs Paraproteinemia
What are proteins induced by vitamin K antagonism (PIVKA)?
Incomletely carboxylated vit K dependednt factors
Due to antagonism, deficiency, absence of vit K
What is used to test PIVKA?
Thrombotest PT
What is a thrombotest PT?
A modified PT test with added FV and fibrinogen so that it detects activity of factors 2, 7, and 10
What does it mean if the thrombotest PT is prolonged?
VitK antagoism or deficiency
Other acquired and hereditary coagulopathies
Cats with hepatic, biliary, or inflammatory bowel diseases
Heparin
What does it mean if there is decreased ATIII activity?
Hypercoagulable states
Decreased production
Loss due to protein-losing nephropathy or severe hemorrhage
Consumption (DIC, spesis, heparin)
What does it mean if there is decreased ATII?
Decreased production
Loss due to protein losing nephropathy
Consumption (DIC, sepsis, heparin)
What is fibrinolysis?
Enzymatic degradation of fibrin
What does fibrinolysis counteract?
Coagulation
When is fibrinolysis initiated?
At the same time as coagulation
Where does fibrinolysis happens?
On the hemostatic plug
What does plasmin do?
Degrades fibrin, fibrinogen, FVa, FVIIIa, vWF, HMWK
What is plasmin inhibited by?
Plasmin inhibitor
What is plasmin cleared by?
Liver
What is plasmin activated by?
tissue-plasminogen activator
What is used to assess fibrinolysis?
Fibrin and fibrinogen degradation products (FDPs)
What are FDPs cleared by?
Lvier
Phagocytes
Kidneys
What are effects of increased FDPs?
Prolonged PT, aPTT, TT, ACT
Impairs platelet function
Compete with fibrinogen for thrombin
Compete with fibrinogen for paltelet binding sites
Associate with fibrin monomers and may disrupt polymerization
What do FDPs detect?
Increased fibrinolysis due to excessive coagualtion
May also detect increased fibrinogenolysis
What is the sample needed to look at FDPs?
Serum: special tubes with thrombin or Botrox atrox venom Citrated plasma (24 hours at 4 deg C)
What are interpretive considerations (false results) of FDPs?
Serum samples: FDPs incorporated into the clots
Generation of FDPs during blood collection
Unspecificty of test
What are interpretive considerations (increased) of FDPs?
Higher concentration in equine neonates Localized intravascular coagualtion DIC Sepsis and inflammatory conditions Hemorrhagic fluid from body cavities Hyperplasminemia leading to increased fibrinolysis
What does D-dimer detect?
Increased fibrinolysis due to excessive coagulation
Factor 13a cross-linked fibrin
What is the sample used for D-dimer?
Citrated plasma
What can biopsy and histology of blood vessels show?
Vasculitis and thrombosis
Diseases involving small vessels
How can you diagnose vessel disorders?
Gross examination
Serology
Imagining
Biopsy
What are causes of large vessel disorders?
Surgical and nonsurgical traumas
Invasion
Aneurysms
Anomalies
What are causes of small vessel disorders?
Vasculitis
Vasculopathies
What are examples of blood disorders?
Impaired primary hemostatsis
Impaired secondary hemostasis
Excessive fibrinolysis: DIC and come envenomations
What is the bleeding pattern seen with a primary hemostasis problem or small vessels?
Petechiae and ecchymosis
Hemorrhages through mucosal surfaces
What is the bleeding pattern seen with a secondary hemostasis problem?
Subcutaneous hematomas and hemorrhage into blood cavities
Hemorrhages through mucosal surfaces
What are some predisposing factors to bleeding disorders?
Breed: might help diagnose inherited diseases
Age: young, prompt inherited
Gender: hemophilia A F8 deficiency and Hemophilia B F9 deficiency, males are not carries only affects; females can be carriers or affected
What is the diagnosis? PT= WRI PTT= WRI FDPs= WRI TT= WRI Platelets= WRI BMBT= Prolonged
Functional platelet problem: vWD or thrombopathia
What is the diagnosis? PT= WRI PTT= WRI FDPs= WRI TT= WRI Platelets= decreased BMBT= Prolonged
Thrombocytopenia with or without platelet functional problems
What is the diagnosis? PT= Prolonged PTT= Prolonged FDPs= WRI TT= Prolonged Platelets= WRI BMBT= WRI or Prolonged
Hepatic disease
What does hepatic disease cause defects in?
Production and clearing of procoagulants, anticoagulants, profibrinolytics, and antifibrinolytics
What tests determine hepatic disease and what is the result?
PT, aPTT, TT, ACT: Decreased production
FDPs and D-dimers: decreased clearance, intravascular coagualtion
Thrombocytopenia: sequestration, decreased production
BMBT: thrombocytopenia, thrombopathia
Fibrinogen, ATIII: decreased production, consumption
What is the diagnosis? PT= WRI PTT= Prolonged FDPs= WRI TT= WRI Platelets= WRI BMBT= WRI
Acquired: hepatic disease, heparin contamination, heparin therapy, VitK antagonism
Inherited: hemophilias A and B F11 deficiency
What is the diagnosis? PT= Prolonged PTT= WRI FDPs= WRI TT= WRI Platelets= WRI BMBT= WRI
Hepatic or heptobiliary disease, VitK antagonism
What is the diagnosis? PT= Prolonged PTT= Prolonged FDPs= WRI TT= WRI Platelets= WRI BMBT= WRI
VitK antagonism, hepatic disease
What is the diagnosis? PT= Prolonged PTT= Prolonged FDPs= Increased TT= Prolonged Platelets= Decreased BMBT= Prolonged
Typical of fulminant consumptive coagulopathy
What is consumptive coagulopathy?
The process of accelerated or unbalanced coagulation that leads to destruction or removal of procoagulant moleucles
DIC
What can you expect to find with consumptive coagulopathy?
3 or more of: Thrombocytopenia Prolonged PT and/or PTT Increased FDPs and/or increased d-dimer Decreased fibrinogen concentration Decreased ATIII RBC fragments
What is dilutional coagulopathy?
Dilution of blood components by colloid fluids, crystalloid fluids, plasma-poor packed erythrocytes, or oxyglobin solution
What can cause the inhibition of functional factors?
Heparin
FDPs
What is thrombosis?
Formation of thrombi within vascular system
Shift of equilibrium between prothrombic factors and antithrombic factors
What are the potential hemostatic abnormailites caused by thrombosis?
Increased D-dimers Increased FDPs Decreased ATIII Thrombocytopenia Increased tt: decreased fibrinogen