Study and analysis of the coagulation factors and their natural inhibitors levels in apheresis derived platelet concentrate units over a five-day storage period

Background: The possibility of utilizing clotting factors in the plasma phase of apheresis platelet concentrates, as a supplement to the standard FFP transfusion for clotting factor replacement needs to be explored. In this study, it was proposed to assess the effect of storage on clotting factors and inhibitors in stored apheresis platelet concentrates. This would give an insight into the hemostatic potential of the plasma phase of the apheresis platelet concentrates.Methods: This study was conducted on a sample size of 45 apheresis platelet concentrate units harvested on Amicus cell separator. Basic coagulation workup along with various coagulation factors and their natural inhibitors were studied in the apheresis platelet concentrates on day ‘0’ and day ‘5’ of the collection.Results: Prothrombin time and activated partial thromboplastin time of the apheresis platelet concentrates was increased on day ‘5’ of the collection but were within the normal range. Fibrinogen, Factor XII, and VWF: Ag showed an increase on day ‘5’ of collection. Protein C, protein S activity, and antithrombin decreased on day ‘5’ of collection. Also, Factors II, VII, IX, X, XI decreased on day ‘5’. The highest fall in activity was seen in the case of Factors V and VIII. Despite the fall, all the clotting factors were maintained within their normal range.Conclusions: Although the activity of most of the coagulation factors showed a decrease, it was maintained within their normal range and efficacy. Therefore, a reasonable hemostatic potential of the clotting factors is expected to be maintained in apheresis platelet concentrates after a storage period of five days at room temperature.

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Increased Activity of Vitamin K-Dependent Clotting Factors in Human Hyperlipoproteinaemia – Association with Cholesterol and Triglyceride Levels

Thrombosis and Haemostasis ◽

10.1055/s-0038-1651852 ◽

1977 ◽

Vol 38 (02) ◽

pp. 0465-0474 ◽

Cited By ~ 46

Author(s):

M Constantino ◽

C Merskey ◽

D. J Kudzma ◽

M. B Zucker

Keyword(s):

Vitamin K ◽

Plasma Lipids ◽

Coagulation Factors ◽

Clotting Factor ◽

Factor X ◽

Clotting Factors ◽

Blood Coagulation Factors ◽

Cholesterol Levels ◽

Type V ◽

Increased Activity

SummaryLevels of blood coagulation factors, cholesterol and triglyceride were measured in human plasma. Prothrombin was significantly elevated in type Ha hyperlipidaemia; prothrombin and factors VII, IX and X in type lib; and prothrombin and factors VII and IX in type V. Multiple regression analysis showed significant correlation between the levels of these plasma lipids and the vitamin K-dependent clotting factors (prothrombin, factors VII, IX and X). Higher cholesterol levels were associated with higher levels of prothrombin and factor X while higher triglyceride levels were associated with higher levels of these as well as factors VII and IX. Prothrombin showed a significant cholesterol-triglyceride interaction in that higher cholesterol levels were associated with higher prothrombin levels at all levels of triglyceride, with the most marked effects in subjects with higher triglyceride levels. Higher prothrombin levels were noted in subjects with high or moderately elevated (but not low) cholesterol levels. Ultracentrifugation of plasma in a density of 1.21 showed activity for prothrombin and factors VII and X only in the lipoprotein-free subnatant fraction. Thus, a true increase in clotting factor protein was probably present. The significance of the correlation between levels of vitamin K-dependent clotting factors and plasma lipids remains to be determined.

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Reduction of salivary tissue factor (thromboplastin) activity by warfarin therapy

Blood ◽

10.1182/blood.v53.3.366.366 ◽

1979 ◽

Vol 53 (3) ◽

pp. 366-374 ◽

Cited By ~ 1

Author(s):

LR Zacharski ◽

R Rosenstein

Keyword(s):

Tissue Factor ◽

Vitamin K ◽

Coagulation Factors ◽

Human Saliva ◽

Factor Vii ◽

Clotting Factor ◽

Activate Factor ◽

Total Protein Content ◽

Factor X ◽

Clotting Factors

Abstract The coagulant of normal human saliva has been identified as tissue factor (thromboplastin, TF) by virtue of its ability to cause rapid coagulation in plasmas deficient in first-stage coagulation factors and to activate factor x in the presence of factor VII and by virtue of the fact that its activity is expressed only in the presence of factor VII and is inhibited by an antibody to TF. The TF is related to cells and cell fragments in saliva. Salivary TF activity has been found to be significantly reduced in patients taking warfarin. The decline in TF activity during induction of warfarin anticoagulation occurs during the warfarin-induced decline in vitamin-K-dependent clotting factor activity, as judged by the prothrombin time. The decrease in TF activity is not related to a reduction in salivary cell count or total protein content or to a direct effect of warfarin on the assay. It is hypothesized that the mechanism by which warfarin inhibits TF activity may be related to the mechanism by which it inhibits expression of the activity of the vitamin-K-dependent clotting factors. Inhibition of the TF activity may be involved in the antithrombotic effect of warfarin.

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Reversible Factor VIII-Abnormalities in Reye’s Syndrome

10.1055/s-0038-1665837 ◽

1979 ◽

Author(s):

J. H. Joist ◽

J. Vargo ◽

M. W. Haymond ◽

J. P. Keating ◽

D. C. DeVivo

Keyword(s):

Factor Viii ◽

Coagulation Factors ◽

Clotting Factor ◽

Metabolic Disturbances ◽

Clotting Factors ◽

Reye's Syndrome ◽

Blood Coagulation Factors ◽

Qualitative And Quantitative ◽

Quantitative Studies ◽

Slow Moving

Reye’s syndrome, an acute and frequently fatal viral infection with severe metabolic disturbances is frequently associated with marked hemostatic alterations. Hypopro-thrombinemia and deficiencies of other clotting factor activities as well as findings suggestive of disseminated intravascular coagulation have been reported. This report deals with detailed sequential qualitative and quantitative studies of blood coagulation factors in four children with this disorder. Depression of factors IX, X, VII, V and II and marked disproportionate elevation of VIIIAHF, VIIIAGN and VIIIvWF associated with a loss of the slow moving component of VIIIAGU on crossed immunoelectro phoresis were consistently observed during the initial phase of the illness. in three of the four children who survived, the changes were fully reversible. Since laboratory evidence for DIC was lacking (normal fibrinogen, consistently negative protamine sulfate precipitation test) the findings seem to indicate that Reye’s syndrome is associated with an acute, transient stimulation of synthesis of normal and abnormal factor VIII as well as decreased synthesis or synthesis of functionally abnormal clotting factors by the liver.

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Properties of Recombinant Human Thromboplastin that Determine Sensitivity to Vitamin K-Dependent Coagulation Factors.

Blood ◽

10.1182/blood.v104.11.533.533 ◽

2004 ◽

Vol 104 (11) ◽

pp. 533-533

Author(s):

Stephanie A. Smith ◽

James H. Morrissey

Keyword(s):

Ionic Strength ◽

Vitamin K ◽

Phospholipid Composition ◽

Coagulation Factors ◽

Factor Vii ◽

Clotting Factor ◽

Sensitivity Index ◽

Clotting Factors ◽

Individual Factor ◽

Factor Deficiency

Abstract Patients undergoing oral anticoagulant therapy (OAT) with coumarins have reduced plasma levels of vitamin K-dependent clotting factors. The primary laboratory test for monitoring OAT is the prothrombin time (PT), in which clotting is initiated by tissue factor (TF). Clotting factors that contribute to the PT, and whose levels respond to OAT, are factor VII (FVII), factor X (FX), and prothrombin, although they are not suppressed to the same extent. Thromboplastin reagents (the source of TF activity in PT tests) can vary dramatically in their sensitivities to the effects of OAT. A calibration system, the International Sensitivity Index (ISI), is widely used to correct the PT for variable thromboplastin sensitivity, but discrepant responses by reagents of similar ISI have been reported. We have undertaken studies aimed at understanding which factors control the sensitivity of thromboplastin reagents, with a goal of creating “designer thromboplastins” whose sensitivities to specific clotting factors can be individually tailored. Thromboplastin reagents were prepared by reconstituting recombinant human TF into phospholipid vesicles containing varying amounts of phosphatidylcholine, phosphatidylserine (PS), and phosphatidylethanolamine (PE). Thromboplastins containing low levels of PS and high ionic strength had the highest sensitivity to OAT (i.e., lowest ISI). PE shifted the dose-response such that lower levels of PS were required to obtain the same ISI value. These studies demonstrate that multiple combinations of phospholipid composition and ionic strength can be used to produce reagents of identical ISI. We hypothesized that reagents of identical ISI values but different composition could have very different responses to changes in the levels of individual coagulation factors. Accordingly, thromboplastin reagents of varying composition were evaluated for their responses to deficiencies of FVII, FX and prothrombin. PT tests were performed using pooled normal plasma mixed with individual factor-depleted plasmas to yield 10%, 3%, 1% or 0.3% of the normal level of the specific clotting factor. Responses of thromboplastin reagents to individual factors were compared by plotting the clotting times obtained with these plasmas on log-log scales versus the percent factor level and fitting lines to the data by linear regression. Interestingly, altering the composition of the thromboplastin reagents dramatically and independently altered their sensitivities to individual clotting factors. For example, increasing ionic strength had no impact on the response to FVII, but markedly enhanced the response to prothrombin deficiency. Furthermore, the effect of changes in ionic strength on specific factors levels differed depending upon the phospholipid composition. These studies demonstrate that thromboplastin reagents of dissimilar composition but nearly identical ISI values can have very different sensitivities to deficiencies in FVII, FX, or prothrombin, so reagents of identical ISI do not necessarily respond to the factor deficiencies induced by OAT in an identical fashion. These studies evaluated samples with isolated individual factor deficiency, whereas patients on OAT have combined factor deficiency and therefore have more potential for discrepancy in PT responses between reagents. Controlling the responsiveness of thromboplastin reagents to deficiencies in individual clotting factors may therefore be desirable for monitoring OAT and for the other clinical diagnostic uses to which PT tests are commonly applied.

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Reduction of salivary tissue factor (thromboplastin) activity by warfarin therapy

Blood ◽

10.1182/blood.v53.3.366.bloodjournal533366 ◽

1979 ◽

Vol 53 (3) ◽

pp. 366-374 ◽

Cited By ~ 15

Author(s):

LR Zacharski ◽

R Rosenstein

Keyword(s):

Tissue Factor ◽

Vitamin K ◽

Coagulation Factors ◽

Human Saliva ◽

Factor Vii ◽

Clotting Factor ◽

Activate Factor ◽

Total Protein Content ◽

Factor X ◽

Clotting Factors

The coagulant of normal human saliva has been identified as tissue factor (thromboplastin, TF) by virtue of its ability to cause rapid coagulation in plasmas deficient in first-stage coagulation factors and to activate factor x in the presence of factor VII and by virtue of the fact that its activity is expressed only in the presence of factor VII and is inhibited by an antibody to TF. The TF is related to cells and cell fragments in saliva. Salivary TF activity has been found to be significantly reduced in patients taking warfarin. The decline in TF activity during induction of warfarin anticoagulation occurs during the warfarin-induced decline in vitamin-K-dependent clotting factor activity, as judged by the prothrombin time. The decrease in TF activity is not related to a reduction in salivary cell count or total protein content or to a direct effect of warfarin on the assay. It is hypothesized that the mechanism by which warfarin inhibits TF activity may be related to the mechanism by which it inhibits expression of the activity of the vitamin-K-dependent clotting factors. Inhibition of the TF activity may be involved in the antithrombotic effect of warfarin.

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VKCFD2 – from clinical phenotype to molecular mechanism

Hämostaseologie ◽

10.1055/s-0037-1617062 ◽

2016 ◽

Vol 36 (S 02) ◽

pp. S13-S20 ◽

Cited By ~ 1

Author(s):

K. J. Czogalla ◽

M. Watzka ◽

J. Oldenburg

Keyword(s):

Vitamin K ◽

Physiological Activity ◽

Coagulation Factors ◽

Clotting Factor ◽

Clotting Factors ◽

Vitamin K Cycle ◽

Factor Deficiency ◽

Er Retention ◽

Deficiency Type ◽

Diagnosis Therapy

SummaryVitamin K 2,3-epoxide reductase complex, subunit 1 (VKORC1) is an enzyme essential for the vitamin K cycle. VKORC1 catalyses the reduction of vitamin K 2,3-epoxide to the quinone form of vitamin K and further to vitamin K hydroquinone. The generated vitamin K hydroquinone serves as substrate for the enzyme γ-glutamyl-carboxylase which modifies all vitamin K-dependent proteins, allowing them to bind calcium ions necessary for physiological activity. Vitamin K-dependent proteins include the coagulation factors FII, FVII, FIX, FX, and proteins C, S und Z. Insufficient VKORC1 enzyme activity results in deficiency of the vitamin K-dependent clotting factors leading to haemorrhagic disorders. This phenotype is known as vitamin K clotting factor deficiency type 2 (VKCFD2). Worldwide, only four families of independent origin have been reported with this rare bleeding disorder. Affected family members carry the mutation VKORC1:p.Arg98Trp in homozygous form, the only mutation found so far to be associated with VKCFD2. Now, ten years after the identification of the VKORC1 gene, the molecular pathomechanism of VKCFD2 has been clarified. The Arg98Trp mutation disrupts an ER retention motif of VKORC1 leading to mislocalisation of the protein to outside the endoplasmatic reticulum. In this review, we summarize the clinical data, diagnosis, therapy and molecular patho -mechanism of VKCFD2.

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Relationships Between Factor XII-Dependent Systems And The “Prorenin”-Renin System In Human Plasma

10.1055/s-0038-1653015 ◽

1981 ◽

Author(s):

E A Wilczynski ◽

A D Purdon ◽

D H Osmond

Keyword(s):

Clotting Factor ◽

Factor Xii ◽

Clotting Factors ◽

In Vitro Techniques ◽

Subsequent Exposure ◽

Control Plasma ◽

Free Plasma

Treatment of plasma with cold (-4°C,72 hr), and with trypsin (0.5 mg trypsin/ml plasma), are well established in-vitro techniques used to activate plasma prorenin. Various clotting factor deficiencies have been found to impair the conversion of prorenin to renin in plasma. Studies with factor XII deficient plasma, in which marked reduction in both cold and tryptic activation was seen, led to further studies on the role of clotting factors and other factor XI I-dependent systems in prorenin activation. Removal of factors II, VII, IX, and X by adsorption onto BaSO4, and subsequent exposure of the residual plasma to cold (-4°C, 48 hr) and trypsin (1 mg/ml), resulted in a decreased capacity for prorenin activation when compared to control plasma, more so in cold than in trypsin-treated plasma. Plasminogen-free plasma responded similarly and, while increased concentrations of trypsin could enhance its prorenin activation to near-normal levels, prolonged cold incubation could not. This suggests that trypsin, added in an appropriate concentration to deficient plasma, may be able to substitute for the missing factor(s), while cold activation is limited by availability of one or more crucial factors. Unmanipulated Fletcher plasma (prekallikrein deficient) has a low level of active renin, and elevated prorenin, symptomatic of a block of prorenin conversion in-vivo. However, cold and tryptic activation were, if anything, relatively greater than normal, especially for trypsin, suggesting that enzymes other than kallikrein are important activators, in-vitro, and can substitute for the missing kallikrein. Thus, neither kallikrein, nor any other single factor studied here, including factor XII, is solely responsible for the activation of plasma prorenin.

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Intracellular maturation of the γ-carboxyglutamic acid (Gla) region in prothrombin coincides with release of the propeptide

Biochemical Journal ◽

10.1042/bj2910723 ◽

1993 ◽

Vol 291 (3) ◽

pp. 723-727 ◽

Cited By ~ 13

Author(s):

R Wallin ◽

C Stanton ◽

S M Hutson

Keyword(s):

Conformational Change ◽

Vitamin K ◽

Secretory Pathway ◽

Coagulation Factor ◽

Coagulation Factors ◽

Coagulation System ◽

Clotting Factor ◽

Clotting Factors ◽

Post Translational Modifications ◽

Carboxyglutamic Acid

Vitamin K-dependent coagulation factors undergo several post-translational modifications before the proteins are secreted into the blood as functional zymogens of the coagulation system. The modifications include Asn-linked glycosylation, Asn/Asp hydroxylation, removal of a signal peptide for translocation of the polypeptide into the endoplasmic reticulum and removal of a propeptide which, when attached to the intracellular coagulation factor precursor, directs the protein for vitamin K-dependent gamma-carboxylation. gamma-Carboxylation of targeted Glu residues results in formation of Ca(2+)-binding gamma-carboxyglutamic acid (Gla) residues. Ca2+ binding by these residues induces a conformational change in the protein which is a necessary event for optimal activation or activity of the clotting factor in blood. In the present study we have monitored the intracellular prothrombin precursor in the secretory pathway of liver cells to determine the effect that the propeptide has on Ca(2+)-dependent folding of the protein. The data provide evidence that the Ca(2+)-induced conformational change required for activation of prothrombin coincides with release of the propeptide in the trans-Golgi apparatus of the liver cell and elucidates an important function for the endoproteinase furin in biosynthesis of vitamin K-dependent clotting factors.

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Reversible Factor VIII-Abnormalities in Reye’s Syndrome.

10.1055/s-0039-1684565 ◽

1979 ◽

Author(s):

J. H. Joist ◽

J. Vargo ◽

M. W. Haymond ◽

J. P. Keating ◽

D. C. DeVivo

Keyword(s):

Factor Viii ◽

Coagulation Factors ◽

Clotting Factor ◽

Metabolic Disturbances ◽

Clotting Factors ◽

Reye's Syndrome ◽

Blood Coagulation Factors ◽

Quantitative Studies ◽

Crossed Immunoelectrophoresis ◽

Slow Moving

Reye’s syndrome, an acute and frequently fatal viral infection with severe metabolic disturbances is frequently associated with marked hemostatic alterations. Hypoprothrombinemia and deficiencies of other clotting factor activities as well as findings suggestive of disseminated intravascular coagulation have been reported. This report deals with detailed sequential qualitative and quantitative studies of blood coagulation factors in four children with this disorder. Depression of factors IX, X, VII, V and II and marked disproportionate elevation of VIIIAHF, VIIIAGN VIIIVWF associated with a loss of the slow moving component of VIIIAGN on crossed Immunoelectrophoresis were consistently observed during the initial phase of the illness. In three of the four children who survived, the changes were fully reversible. Since laboratory evidence for DIC was lacking (normal fibrinogen, consistently negative protamine sulfate precipitation test) the findings seem to indicate that Reye's syndrome is associated with an acute, transient stimulation of synthesis of normal and abnormal factor VIII as well as decreased synthesis or synthesis of functionally abnormal clotting factors by the liver.

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Thrombotic activation before and after total hip arthroplasty. A prospective cohort study

BMC Musculoskeletal Disorders ◽

10.1186/s12891-021-04566-1 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Marta Burbul ◽

Dariusz Tomaszewski ◽

Anna Rogalska ◽

Krzysztof Gawroński ◽

Sławomir Literacki ◽

...

Keyword(s):

Total Hip Arthroplasty ◽

Blood Loss ◽

Factor Viii ◽

Hip Arthroplasty ◽

Clotting Factor ◽

Diagnostic Study ◽

Fibrinogen Concentration ◽

Clotting Factors ◽

Normal Range ◽

Total Hip

Abstract Background Total hip arthroplasty (THA) causes acute blood loss. It may lead to a deficiency in coagulation factors, which, in turn, may lead to increased bleeding during the postoperative period. Methods Thirty patients (18 women) with a mean age of 67 years (range: 63–72 years) participated in this prospective diagnostic study. THA was performed without tranexamic acid administration in the perioperative period. Activities of clotting factors II, VIII, X, and fibrinogen concentration were evaluated before surgery, 6 hours after the procedure, 2, 4, and 6 days after the operation. All laboratory tests were performed using ACL TOP 500 CTS analyzer. Results No thromboembolic complications were noted during hospitalization. Mean fibrinogen concentration was 366 mg/dL before surgery, which decreased to 311 mg/dL 6 hours after the operation and peaked at 827 mg/dL on the 4th day after the procedure. Activities of factors II and X decreased on the second and fourth days after surgery. Although the activity of factor VIII decreased after the procedure, it remained within the normal range. Increased baseline fibrinogen concentrations were observed in 6 out of 30 (20%) patients. Mean blood loss was 1332 mL (range, 183–2479 mL) and did not correlate with changes in clotting factor activities. Conclusions In patients undergoing THA, fibrinogen acts as an acute-phase protein. Activities of clotting factors II and X normalize within 6 days, and although the activity of factor VIII decreases, it remains within the normal range. Trial registration The study was pre-registered May 1st, 2020 on ClinicalTrials.gov

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