scholarly journals Evaluation of Thrombin Generation Assay in Patients with Hemophilia

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5061-5061
Author(s):  
Mehran Karimi ◽  
Sezaneh Haghpanah ◽  
Asghar Bazrafshan ◽  
Peyman Eatemadfar ◽  
Javad Dehghani ◽  
...  
Keyword(s):  
Disease Severity ◽  
Thrombin Generation ◽  
Lag Time ◽  
Activity Level ◽  
Screening Tests ◽  
Control Group ◽  
Factor Activity ◽  
Thrombotic Risk ◽  
Plasma Factor ◽  
Bleeding Score

Abstract Background: Diagnosis of hemophilia is based on detection of factor level and coagulation screening test. These screening tests can not reflect global hemostatic balance. Thrombin generation (TG), as a focal point and key process, is an important step in hemostasis and thrombosis. Recent evidence has indicated that TG assay is a better predictor of the coagulation capacity and subsequently overall assessment of hemostasis compared with traditional coagulation tests. The parameters of the thrombogram are helpful in the evaluation of bleeding or thrombotic risk and also in the management of these situations. Objective: The aim of this study is to evaluate the correlation between TG parameters with bleeding symptoms and disease severity in patients with hemophilia. Methods: In this cross-sectional study, 59 male patients with hemophilia were randomly selected from those already registered at Shiraz Hemophilia Center of southern Iran, from February to December 2012. Also, 38 healthy age-matched men were considered as a control group from those referred for check up. Informed written consent was obtained from all participants. The proposal was approved by the Medical Ethics Committee of Shiraz University of Medical Sciences. Disease severity was defined based on the activity level of deficient factor (severe <1%, moderate from 1 to 5%, and mild >5% IU/dL). Bleeding score (BS) was calculated by performing a clinical evaluation using a modified questionnaire based on Tosetto et al- questionnaire. Correlation of TG parameters with disease severity and BS were determined. Results: From 59 patients with hemophilia (52 hemophilia A and 7 hemophilia B), 40 patients (68%) had severe disease, 14 (24%) moderate and 5 (8%) mild disease. All TG parameters showed statistically significant differences between patients and controls (P<0.001). Only Lag time showed a statistically significant correlation with BS (rs= 0.316, P =0.016). All TG parameters except peak showed association with disease severity (P<0.05). In addition, endogenous thrombin potential showed a significant correlation with factor activity level (rs= 0.459, P <0.001). Both Lag time and start tail showed significant negative correlations with factor activity level (rs= -0.488, P <0.001 and rs=- 0.289, P <0.026 respectively). Conclusion: Based on our results, although most of the TG parameters evaluated were not significantly correlated with bleeding score of hemophilia patients, the majority of TG parameters were significantly associated with factor activity level and disease severity. In patients with hemophilia, plasma factor activity level has a poor predictive value in the evaluation of efficacy of treatment; but it seems that TG assay is an appropriate tool for assessment of global hemostasis and better reflection of clotting function in the management of patients with hemophilia. Disclosures No relevant conflicts of interest to declare.

scholarly journals Impact of Adopting Population Pharmacokinetics for Tailoring Prophylaxis in Haemophilia A Patients: A Historically Controlled Observational Study

2019 ◽  
Vol 119 (03) ◽  
pp. 368-376 ◽  
Author(s):  
Michaela Stemberger ◽  
Felix Kallenbach ◽  
Elisabeth Schmit ◽  
Alanna McEneny-King ◽  
Federico Germini ◽  
...  
Keyword(s):  
Clinical Practice ◽  
Activity Level ◽  
Critical Threshold ◽  
Life Questionnaire ◽  
Haemophilia A ◽  
Factor Activity ◽  
Traffic Light ◽  
Plasma Factor ◽  
Population Pk ◽  
The Impact

Background Performing individual pharmacokinetics (PK) studies in clinical practice can be simplified by adopting population PK-based profiling on limited post-infusion samples. The objective of this study was to assess the impact of population PK in tailoring prophylaxis in patients with haemophilia A. Patients and Methods Individual weekly treatment plans were developed considering predicted plasma factor activity levels and patients' lifestyle. Patients were trained using a visual traffic-light scheme to help modulate their level of physical activity with respect to factor infusions timing. Annualized joint bleeding rate (ABJR), haemophilia-specific quality of life questionnaire for adults (Haemo-QoL-A) and factor utilization were measured for 12 months before and after tailoring, compared within patients and analysed separately for those previously on prophylaxis (P), situational prophylaxis (SP) or on-demand (OD). Results Sixteen patients previously on P, 10 on SP and 10 on OD were enrolled in the study. The median (lower, upper quartile) ABJR changed from 2.0 (0, 4.0) to 0 (0, 1.6) for P (p = 0.003), from 2.0 (2.0, 13.6) to 3.0 (1.4, 7.2) for SP (p = 0.183) and from 16.0 (13.0, 25.0) to 2.3 (0, 5.0) for OD (p = 0.003). The Haemo-QoL-A total score improved for 58% of P, 50% of SP and 29% of OD patients. Factor utilization (IU/kg/patient/year) increased by 2,400 (121; 2,586) for P, 1,052 (308; 1,578) for SP and 2,086 (1,498; 2,576) for OD. One of 138 measurements demonstrated a factor activity level below the critical threshold of 0.03 IU/mL while the predicted level was above the threshold. Conclusion Implementing tailored prophylaxis using a Bayesian forecasting approach in a routine clinical practice setting may improve haemophilia clinical outcomes.

Kinetics of Thrombin Generation in Patients with Multiple Myeloma.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3535-3535 ◽  
Author(s):  
Anna D. Petropoulou ◽  
Grigoris T. Gerotziafas ◽  
Kostas Zervas ◽  
A. Mpanti ◽  
Michel Meyer Samama ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Endothelial Cell ◽  
Plasma Levels ◽  
Thrombin Generation ◽  
Cell Damage ◽  
Control Group ◽  
Endothelial Cell Damage ◽  
Thrombotic Risk ◽  
Soluble Thrombomodulin ◽  
Significant Difference

Abstract Thalidomide has emerged as a promising treatment for multiple myeloma (MM). Thrombosis is the most serious complication of thalidomide therapy, essentially when it is combined with dexamethasone. The pathogenesis of thrombosis in MM patients (pts) treated with thalidomide is not clear and probably of multi factorial origin. We used the Thrombin Generation test (TGT) and measured the plasma levels of soluble thrombomodulin (sTM) to better clarify the MM-related and thalidomide-related thrombogenicity. TGT was performed in citrated frozen platelet poor plasma (PPP). Blood was obtained from 26 MM pts, Salmon and Durie stage II and III, 62.5 years old (42–77), 9 males and 17 females, 10 treated with thalidomide (100–200mg/d orally) and dexamethasone (40mg/d for 4 days) (TD group) and 16 receiving no treatment (MM group). 13 healthy volunteers formed the control group. Thrombin Generation (TG) was initiated by adding the PPP reagent (Thrombogram-Thrombinoscope®) and the triggering solution (CaCl2 and fluorogenic substrate). We analyzed the endogenous thrombin potential (ETP), the Cmax and the velocity index of TG. The plasma levels of sTM in PPP were measured by a specific ELISA (Diagnostica Stago, France). In the MM group we observed an increase of the ETP, though not significant compared to the controls. The Cmax was almost equal to the control group value, while the velocity index of TG was statistically lower in the MM group compared to controls. In the TD group, a statistically significant increase of ETP was observed as compared to the control group. The Cmax was higher, compared to controls, though not significantly, whereas the velocity index of TG was almost equal to the control group value. There was no significant difference in the TG parameters between MM and TD groups. sTM in the control group was 45±14ng/ml. Both groups of pts had significantly increased sTM plasma levels as compared to the control but the difference between the two groups did not reach significance. Results are shown in Table 1. In patients with MM coexists an increase of sTM, a marker of endothelial cell damage, together with an increased TG capacity. The addition of thalidomide treatment is associated with a slight but not significant increase of ETP and Cmax. The co-existence of endothelial cell damage with increased TG capacity could be associated to the increased thrombotic risk in MM patients treated with thalidomide. This hypothesis will be controlled in a prospective study. Table 1: Thrombogram parameters and sTM plasma levels of studied pts. Control MM group TD group * Results significantly different between the MM and TD groups and the control group (p<0.05 vs the control group) ETP (nM×min) 1399±297 1651±478 1747±448* Cmax (nM) 366±54 342±52 402±99 Velocity Index (nM/min) 198±45 160±19* 184±65 STM (ng/ml) 45±14 84±42* 73±30*

Increased Microparticle-Linked Procoagulant Activity In Patients with Primary Immune Thrombocytopenia.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3707-3707
Author(s):  
Elena G. Arias Salgado ◽  
Ihosvany Fernández Bello ◽  
Mayte Álvarez Román ◽  
Isabel Rivas ◽  
Mónica Martín Salces ◽  
...  
Keyword(s):  
Platelet Count ◽  
Immune Thrombocytopenia ◽  
Thrombin Generation ◽  
Conflicts Of Interest ◽  
Lag Time ◽  
Peak Height ◽  
Procoagulant Activity ◽  
Control Group ◽  
Increased Risk ◽  
Primary Immune Thrombocytopenia

Abstract Abstract 3707 Primary immune thrombocytopenia (ITP) is an acquired immune-mediated disorder characterized by isolated thrombocytopenia (platelet count less than 100,000/μL) and the absence of any obvious initiating and/or underlying cause for the thrombocytopenia. In spite of the low platelet number, some thrombocytopenic patients seldom bleed, indicating the existence of other factors that regulate haemostasis in these patients. Elevated levels of plasma microparticles (MPs) had been observed in IPT patients. MPs are vesicles with a size less than 0.5 micrometers, derived from cell membranes after their activation or apoptosis. Most MPs are highly procoagulant, expressing annexin V binding sites and tissue factor. However, relatively little is known of their specific functions in ITP. In the present study we aim to elucidate if a relationship exists between microparticle-linked procoagulant activity and haemostasis in ITP patients. Twenty-two ITP patients, 3 male and 19 female, aged between 25 to 92 years, were included. Sixteen age- and sex-matched healthy individuals were used as control group. Platelet-related primary haemostasis was evaluated with an automated platelet function analyzer (PFA-100®, Siemens Healthcare Diagnostics). Samples of citrated blood were aspirated under a shear rate of 4,000–5,000/s through a 150-micrometer aperture cut into a collagen-ADP (COL-ADP) or collagen-epinephrine (COL-EPI) coated membrane. The platelet haemostatic capacity is indicated by the time required for the platelet plug to occlude the aperture (closure time), which is expressed in seconds. MP procoagulant activity was determined with ZYMUPHEN MP-Activity kit (Aniara, Mason, Ohio) and by calibrated automated thrombography (CAT) in plasma samples obtained after 2 centrifugations at room temperature (first: 15 min at 1,500 g, second: 2 min at 13,000 g). These methods measure endogenous thrombin generation. CAT evaluates four parameters of thrombin generation: the endogenous thrombin potential (ETP), lag time, time to peak (TTP) and peak height. PFA-100® determinations with COL-EPI and COL-ADP cartridges in blood samples from ITP patients with less than 50,000/μL showed longer closure times than control group (p<0.05), whereas samples from ITP patients with a platelet count between 50,000/μL and 100,000/μL showed closure times of the same order of magnitude as control ones (platelet count ranging from 162,000 to 368,000)μL).Plasma from these patients had higher MP-mediated procoagulant activity evaluated with ZYMUPHEN kit (control 6.1+3.9 nM, ITP group 10,1±8.2 nM, p<0.05) as well as with CAT (ETP (nM*min): control: 1692.6±341.9, ITP: 2191,8±398.9, p<0.01; lag time (min): control: 19.9±8.2, ITP: 14.3±4.3, p<0.05; TTP (min): control: 22.0±8.3, ITP:16.3±4.4, p<0.05; peak height (nM): control: 389.7±70.6, ITP: 498,8±97.5, p<0.01). Our results indicate that increased MP procoagulant activity in ITP patients may be protective against bleeding events that should be observed in those thrombocytopenic conditions. Three of the ITP patients included in this study had been splenectomyced and we consider of interest to point out that two of them in spite of recovering a normal platelet count still maintain a high MP procoagulant activity. This observation agrees with a recent work that postulates that MPs might contribute to an increased risk of thrombosis, progression of atherosclerosis and cardiovascular disease following splenectomy (Fontana et al, Thromb Research, 2008;122:59). Disclosures: No relevant conflicts of interest to declare.

Procoagulant State in Current and Former Anabolic Androgenic Steroid Abusers

2018 ◽  
Vol 47 (04) ◽  
pp. 647-653
Author(s):  
Jon Rasmussen ◽  
Mikkel Frandsen ◽  
Morten Schou ◽  
Marie Johansen ◽  
Jens Faber ◽  
...  
Keyword(s):  
Linear Regression ◽  
Protein C ◽  
Thrombin Generation ◽  
Protein S ◽  
Lag Time ◽  
Anabolic Androgenic Steroid ◽  
Thrombotic Risk ◽  
Time To Peak ◽  
Androgenic Steroid ◽  
Cardiovascular Morbidity And Mortality

Background Anabolic androgenic steroid (AAS) abusers are considered at increased risk of cardiovascular morbidity and mortality. We hypothesized that current and former AAS abuse would induce a procoagulant shift in the haemostatic balance. Methods Men 18 to 50 years of age were included as current AAS abusers, former AAS abusers or controls. Morning blood samples were collected after overnight fasting. Thrombin generation (lag time, time to peak, peak height, and endogenous thrombin potential [ETP]) and coagulation factor II (prothrombin), VII and X, antithrombin, protein C, free protein S and tissue factor pathway inhibitor (TFPI) were assessed. Groups were compared by ANOVA or Kruskal–Wallis test and probabilities were corrected for multiple comparisons. Associations were evaluated using linear regression models. Results ETP was increased around 15% in current (n = 37) and former (n = 33) AAS abusers compared with controls (n = 30; p < 0.001). Prothrombin and factor X were increased ≥10% in AAS abusers and prothrombin was a predictor of ETP (p < 0.0005). Lag time and time to peak were increased 10 to 30% in current AAS abusers (p < 0.001) and associated with higher concentrations of TFPI, antithrombin, protein C and protein S (p < 0.0005; = 0.005). Multivariate linear regression, with all coagulation inhibitors as covariates, identified TFPI to be independently associated with lag time and time to peak (p < 0.0005). Conclusion Thrombin generation is augmented in current and former AAS abusers, reflecting a procoagulant state, with altered concentrations of coagulation proteins. Prospective studies are needed to clarify whether these findings translate into an increased thrombotic risk in AAS abusers potentially even after cessation.

Thrombin Generation In Patients with Essential Thrombocythemia Treated with Anagrelide

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 5050-5050
Author(s):  
Krystyna M Zawilska ◽  
Agnieszka Skubiszak ◽  
Jolanta M Kurosz ◽  
Jerzy Jaworski
Keyword(s):  
Essential Thrombocythemia ◽  
Thrombin Generation ◽  
Lag Time ◽  
Control Group ◽  
Study Group ◽  
Time To Peak ◽  
History Of ◽  
Wbc Count ◽  
Age Range ◽  
Von Willebrand

Abstract Abstract 5050 Introduction: Thrombosis may be serious and life threatening in patients with essential thrombocythemia (ET). Bleeding is usually from the gastrointestinal tract, and is, in most cases, generally associated with a platelet count greater than 1 million/μL. The mechanism by which thrombocythemia produces hemorrhage or thrombosis is not well defined. Several defects have been described, including a decrease in platelet aggregation or hyperaggregation, a decrease in von Willebrand ristocetin cofactor activity and high molecular weight von Willebrand factor multimers. Some reports show patients with an acquired deficiency of antithrombin, protein C, and acquired APC resistance, probably due to a reduction in free protein S levels. More recently the presence of JAK2V617F mutation and baseline leukocyte (WBC) count have been considered as independent predictors of major thrombosis in ET. Aim of the study: To assess the thrombin generation by the calibrated automated thrombogram method in plasma from ET patients treated with anagrelide. This assay reflects the net results of the procoagulant and anticoagulant forces operating in plasma. The area under the thrombin generation curve (also known as the endogenous thrombin potential (ETP) is a good overall indicator of prothrombotic and hemorrhagic tendency. It could be hypothezised, that platelet-lowering treatment could diminish haemostasis disturbances in ET. Material and methods: The study group consisted of ten ET patients (4 males and 6 females; age range, 24–70 years), diagnosed according to the Polycythemia Vera Study Group criteria. All patients have been treated with anagrelide for mean 4 years (range 1 – 7 years), without antiplatelet drugs. At the time of enrollment their mean platelet number was 344 000/μL (210 000 – 535 000), WBC count 8 600/μL (3 300 – 9 800), and hematocrit 38 (33-47). One patient had a history of splenic vein thrombosis, another had a gastrointestinal bleeding before the start of treatment. Four ET patients (40%) were positive for the JAK2V617F. Ten healthy subjects (4 males and 6 females; age range, 30–70 years) without history of thrombohemorrhagic events, acted as a control group. Thrombin generation (TG) was determined by calibrated automated thrombography (Technothrombin TGA-Technoclone). Thrombin generation curves were described in terms of lag time, peak height, time to peak, slope and ETP. Whole blood thromboelastometry with TEM-ROTEM delta - Pentapharm GmbH has been performed as well. Results: Thrombin generation (ETP) in platelet free plasma was significantly increased in patients with ET (2235 ± 376 nM/min) in comparison with controls (1631 ± 257 nM/min; p=0,0082), the TG lag time, peak, time to peak and slope were also abnormal. Patients with ET showed markedly increased values of INTEM coagulation time-CT, clot formation time-CFT and increased maximum clot firmness, when compared to results of the control group (Table). There were no differences in the EXTEM values. Conclusion: Our results suggest that patients with essential thrombocythemia, in spite of a long-lasting anagrelide treatment, still exhibit an increased thrombin generation as well as formation of a clot with an increased firmness. Disclosures: No relevant conflicts of interest to declare.

Thrombotic risk assessment in the antiphospholipid syndrome requires more than the quantification of lupus anticoagulants

Blood ◽  
2010 ◽  
Vol 115 (4) ◽  
pp. 870-878 ◽  
Author(s):  
Katrien Devreese ◽  
Kathelijne Peerlinck ◽  
Marc F. Hoylaerts
Keyword(s):  
Risk Assessment ◽  
Thrombin Generation ◽  
Lag Time ◽  
Peak Height ◽  
Factor Vii ◽  
Thromboembolic Complications ◽  
Thrombotic Risk ◽  
Predictive Values ◽  
Lupus Anticoagulants ◽  
Glycoprotein I

Abstract Lupus anticoagulants (LACs) are associated with thromboembolic complications (TECs). LACs can be detected by their anticoagulant properties in thrombin generation assays, by the peak height (PH) and lag time (LT). To assess the thrombotic risk in LAC-positive patients, we have expressed the LAC activity quantitatively by PH/LT calibration curves, constructed for mixtures of monoclonal antibodies against β2-glycoprotein I (β2GPI) and prothrombin, spiked in normal plasma. PH/LT was determined in LAC patients, with (n = 38) and without (n = 21) TECs and converted into arbitrary LAC units. LAC titers ranged from 0 to 200 AU/mL, with 5 of 59 patients being negative. In the positive LAC titer population (54 of 59), LAC and anti-β2GPI immunoglobulin G (IgG) titers correlated with TECs, with odds ratios of 3.54 (95% CI, 1.0-1.7) and 10.0 (95% CI, 1.98-50.6), respectively. In patients with single or combined low titers, useful predictions on thrombosis could be made only after additional measurements of soluble P-selectin and factor VII. This layered strategy yielded positive and negative predictive values, sensitivity, and specificity values approximately 90% in this subgroup. Hence, LAC and anti-β2GPI IgG titers, when combined with selected markers of the hypercoagulable state, allow a relevant thrombotic risk assessment in nearly all patients with LACs.

scholarly journals Global Coagulation Assays in the Normal Population: Female Gender, Older Age and East Asian Ethnicity Associated with Prothrombotic Parameters

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 4678-4678 ◽  
Author(s):  
Prahlad Ho ◽  
Hui Yin Lim ◽  
Cheryl Ng ◽  
Carole L. Smith ◽  
Geoffrey Donnan ◽  
...  
Keyword(s):  
Thrombin Generation ◽  
Normal Population ◽  
East Asian ◽  
Lag Time ◽  
Peak Height ◽  
Clot Lysis ◽  
Older Individuals ◽  
Thrombotic Risk ◽  
Coagulation Assays ◽  
Α Angle

Abstract Aim Thrombotic diseases are major causes of morbidity and mortality, and yet there are currently no laboratory tests to evaluate this risk. Global coagulation assays such as thrombin generation via calibrated automated thrombogram (CAT) and thromboelastography (TEG), may be better surrogate measures of an individualÕs thrombosis risk. However, the evaluation of age, race and gender differences in the normal population is important prior to the inclusion of these assays into studies on diseased populations. Methods Normal controls with no thrombotic history and no anticoagulant/antiplatelet use were recruited. Routine tests were performed to exclude underlying risk factors including full blood count, thrombophilia screen and fasting lipids. All samples were citrated; platelet poor plasma (PPP) samples were double-centrifuged at 2500G, aliquoted and frozen at -80o C within 2 hours of collection. TEG 5000S analysis was performed within 4 hours of collection, using whole citrated blood under standard manufacturer guidelines. CAT was performed with PPP using standard commercially available (STAGO) 5 pmol reagent and flurogenic (Fluca-Kit). Results 63 normal controls (43F, 20M) with median age of 36.5 (20-75) years were recruited. None had abnormal platelet or coagulation parameters. TEG: 36 volunteers (57%) had TEG parameters outside the manufacturerÕs normal range. Females and older individuals (age >50 years) had more prothrombotic TEG parameters including increased maximum amplitude (MA), alpha-angle and derived thrombin generation; with reduced R and K time. Older females (age >50 years) also had more prothrombotic parameters compared to younger females. Clot lysis (LY30) was significantly lower in the older population with no gender differences. (Table 1) CAT: Females had higher endogenous thrombin potential (ETP), which represented the total thrombin formed, and higher velocity index. Older individuals had higher ETP but paradoxically a longer lag time. (Table 1) CAT vs TEG: There was no correlation between TEG parameters (such as MA or derived thrombin generation), and ETP via CAT (r2=0.005 and r2=0.006 respectively). Race differences: Of the 63 volunteers, 25 and 30 were of European and East Asian origin respectively. East Asians had significantly higher ETP, thrombin peak and velocity index (p<0.01), but these differences were less apparent using TEG, which also evaluated the platelet effect on clot formation. Statistically significant variation was only seen in the α-angle (p=0.04). (Table 2) Conclusion This study demonstrates that there are significant gender and age differences within the normal population. 57% of our volunteers had at least one TEG parameter outside the manufacturers reference range. Females had more prothrombotic parameters, which was independent of hormonal status. Older individuals (age >50 years) having more prothrombotic parameters, particularly clot lysis (LY30) using TEG, suggests a role of fibrinolysis in the futher evaluation of thrombotic risk. East Asians had significantly lower CAT parameters, which may explain the lower venous thrombotic risk seen in this population. Our results suggest that a localized reference range, tailored to gender, age and ethnicity is required for global coagulation assays. Table 1. Gender and age difference with TEG and CAT Male Female Age <50years Age ³50years No of controls 20 43 32 31 TEG parameters R time (min) 8.7 6.8 p <0.01 8.2 6.6 p =0.01 K time (min) 3.2 2.2 p <0.01 2.9 2.2 p =0.03 α-angle (¡) 50.6 59.8 p <0.01 52.2 61.3 p <0.01 MA (mm) 54.3 59.7 p <0.01 56.1 59.8 P=0.02 LY30 (%) 1.8 1.9 p=0.92 3.1 0.7 p <0.01 Derived Thrombin Gen 671 724 p <0.01 676 737 p <0.01 CAT parameters Lag Time (min) 3.0 3.2 p=0.53 2.8 3.4 p <0.01 ETP (nM/min) 1245 1399 p =0.03 1290 1403 p=0.06 Peak height (nM) 211 245 p=0.06 221 243 p=0.13 Vel Index (nM/min) 65 86 p =0.03 79 77 p=0.97 Time to Peak (min) 6.5 6.4 p=0.58 6.2 6.8 p=0.06 Table 2. Ethnic difference with CAT and TEG parameters Manufacturer values East Asian (n=30) European (n=25) p-value (% variation) TEG parameters R time (min) 2-8 7.8 6.8 0.15 (+14%) K time (min) 1-2 2.8 2.2 0.06 (+23%) α-angle (¡) 55-78 53.9 60.0 0.04 (-10%) MA (mm) 51-69 57.4 58.4 0.57 (-2%) Lysis time, LY30 (%) 0-8 1.9 2.4 0.53 (-34%) CAT parameters Not available Lag Time (min) 3.0 3.2 0.22 (-7%) ETP (nM/min) 1231.5 1411.7 <0.01 (-23%) Peak height (nM) 206.8 254.8 <0.01 (-19%) Vel Index (nM/min) 65.6 92.5 <0.01 (-30%) Disclosures No relevant conflicts of interest to declare.

Stability Of Thrombin Generation In Cirrhotic Patients

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2361-2361
Author(s):  
Thomas Sinegre ◽  
Armand Abergel ◽  
Géraldine Lamblin ◽  
Marc G Berger ◽  
Aurélien Lebreton
Keyword(s):  
Protein C ◽  
Thrombin Generation ◽  
Activated Protein C ◽  
Final Concentration ◽  
Control Group ◽  
Healthy Controls ◽  
Thrombotic Risk ◽  
Compensated Cirrhosis ◽  
Cirrhotic Patients ◽  
The Stability

Abstract Introduction Cirrhotic patients have a significant impairment of hemostasis. Most procoagulant and anticoagulant factors decrease but there are still exceptions such as factor VIII. Since many years, new approaches allowed to consider the cirrhotic patients as patients exposed to a thrombotic risk in many circumstances. In this concern, global tests as Thrombin Generation Assay (TGA) have an interest and many studies using thrombomodulin (TM) demonstrated a resistance to activated protein C (aPC) pathway. These assays evaluate the variations of both procoagulant factors and anticoagulant factors. However, the stability of thrombin generation in these patients is unknown. The aim of this study was to evaluate the stability of thrombin generation parameters (with and without TM and aPC) in cirrhotic patients compared to healthy volunteers. Patients and Methods 8 cirrhotic patients and 15 healthy controls were included in this study. For each patient, the follow-up period covers three months including three blood samples (D0, W6 and W12). Patients included in the study are cirrhotic (Prothrombin Time < 70% and / or liver dysmorphia and / or Fibroscan> 20 kPa and / or histology and / or association of portal hypertension and liver failure) of alcoholic origin. They are exclusively male, free of hepatocellular carcinoma and not anticoagulated. Thrombin Generation was performed in platelet-poor plasma in the presence / absence of TM (3.6 nmol/L) and in the presence/absence of aPC (1 nmol/L). The thrombin generation test was carried out according to the principle described by Hemker using Fluoroskan Ascent®, and the analysis software ThrombinoscopeTM. Reagent contains phospholipids (4 microM / L) and tissue factor (5 pM/L). TM is soluble thrombomodulin purified from rabbit lungs at final concentration 3.6 nmol/L. Human aPC was used at final concentration of 1 nmol / L (IC 50). The results are expressed as ratios: with and without TM and with and without PCa. Lag Time (LT), Endogenous Thrombin Potential (ETP), Peak Height (PH) and Time to Peak (TP) were analyzed. Statistical analysis compared the coefficient of variation (CV) between the control group and cirrhotic patients for each ratio (with and without TM with and without PCa) and that for each parameter by the Mann-Whitney test. Results Patients included in the study have chronic liver disease. Seven of them have compensated cirrhosis (CHILD A) and one cirrhotic patient has complicated cirrhosis (infection). The average age of cirrhotic is 50 (range 37-57). For both TM or aPC, all parameters of TGA are stable in cirrhotic patients compared with healthy controls. Regarding ETP, the most used parameter, CV of the ratio with and without TM is 12% (2-24) for the controls and 13% (2-22) for the cirrhotic patients while CV of the ratio with and without aPC is 11% (3-24) for the controls and 11% (6-21) for the cirrhotic patients. Similar matches were found for PH: 12% (3-25) for the controls versus 12% (1-22) for the cirrhotic patients with TM and 12% (1-20) for the controls versus 11% (5-19) for the cirrhotic patients. Similar results were found for LT and TP. Conclusions The Thrombin generation - performed with Thrombomodulin and activated Protein C and expressed with a ratio - in patient with cirrhosis is similarly stable than the thrombin generation in the control population. These observations can be used in the management and the design of further studies involving cirrhotic patients. Now, it is possible to follow the cirrhotic patients with TGA in order to detect the occurrence of a hypercoagulability state. However patients included in the study were mostly compensated cirrhosis. Further studies are needed to evaluate the stability of thrombin generation in the other subgroups of cirrhotic patients. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Decreased Thrombin Generation Potential in Lymphoma Patients Is Associated with Increased D-Dimer, CRP, Vwf and TNF-α. Interrelationship between Thrombogenesis and Inflammation

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5239-5239
Author(s):  
Fakiha Siddiqui ◽  
Emily Bontekoe ◽  
Darko Antic ◽  
Debra Hoppensteadt ◽  
Grigoris Gerotziafas ◽  
...  
Keyword(s):  
Hodgkin Lymphoma ◽  
Thrombin Generation ◽  
Inflammatory Responses ◽  
Sandwich Elisa ◽  
Lag Time ◽  
Inflammatory Biomarkers ◽  
Activation Process ◽  
Thrombotic Risk ◽  
Tnf Α ◽  
D Dimer

Introduction: The prevalence of thrombosis in lymphoma patients is reportedly high and ranges from 3 - 10%, and further increased at advanced stages of the disease especially in hgNHL. Inflammation and other vascular factors contribute to the pathogenesis of these thrombotic complications. Biomarkers of hemostatic activation, vascular dysfunction and inflammation are elevated in lymphoma. We have previously reported that thrombin generation biomarkers such as D-Dimer, thrombin anti-thrombin complex (TAT) and prothrombin fragment (F1.2) along with inflammatory biomarkers are increased in cancer patients. Interestingly, despite an increase in thrombin generation markers, thrombin generation potential in these patients is decreased. This study was designed to compare the thrombin generation potential and its relevance to the generation of various biomarkers of hemostatic activation process and inflammatory responses. Methods: Citrated blood samples from 90 patients with confirmed diagnosis of non-Hodgkin lymphoma (NHL), Hodgkin lymphoma (HL) and Chronic lymphocytic leukemia/Small lymphocytic lymphoma (CLL/SLL) were collected from the Clinic of Hematology Unit, University of Belgrade, Belgrade, Serbia. 50 samples of normal human plasma (NHP) was obtained from George King Biomedical (Overland park, KS). NHP was prepared for referencing purposes. Thrombin generation studies were carried out using a commercially available a kinetic fluorogenic substrate method (calibrated automated thrombogram; CAT). Thrombin generation parameters such as peak thrombin (PT), lag time (LT) and area under the curve (AUC) were compiled. Plasma levels of D-Dimer, CRP, vWF and TNF- α were measured by using commercially available Sandwich ELISA assays. Results were compiled in terms of mean ± SD. Correlation analysis carried out by using Prism, Graphpad. Results: On a cumulative basis, lymphoma patients showed an increase in lag time (2.9 1.17) in comparison to the NHP (2.7). The peak thrombin levels was decreased (119.0 46.2) in the lymphoma patients in comparison to NHP (159.7 31.1). The AUC was decreased (670.2 290.01) in the lymphoma patients in comparison to NHP (700.9 ). When the lymphoma patients were sub grouped, peak thrombin levels were similar in HL (129.9), NHL (114.7) and CLL (127.4) and were decreased compared to NHP (159.7). However the AUC was increased in the HL (769), decreased in NHL (642.4) and was comparable for CLL (715.4) compared to NHP (700.9). Variations in lag time were noted in the three groups. The lymphoma patients also exhibited elevated levels of D-Dimer (878 1205 ng/ml) in comparison to NHP (280 ng/ml), CRP (15.2 26.6 ug/ml) in comparison to NHP (0.9 ug/ml), vWF (271.8 189.7 %) in comparison to NHP (122%) and TNF-α (16.4 20.64 pg/ml) in comparison to NHP (4.2 pg/ml). Interestingly, when the lymphoma group were subdivided into various groups, the D-Dimer level was highest in HL following the order HL>NHL>CLL. CRP and vWF levels were highest in HL and comparable in both NHL and CLL. Wide scatter in TNF-α data was noted with CLL exhibiting highest levels with the ranked order CLL>NHL>HL. The composite data on thrombin generation parameters and biomarkers is shown on the table 1A. In the HL group D-Dimer and CRP correlated well with AUC and PT, in this group vWF also showed a good correlation with PT. In the NHL group, CRP correlated well with LT. In the CLL group none of the biomarkers correlated well with the thrombin generation parameters. On a cumulative basis, D-Dimer, CRP, vWF and TNF-α had no significant relevance to thrombin generation parameters. The composite regression coefficience data is shown on table 1B. Conclusion: Lymphoma patients represent a heterogenous group in which both the hypercoagulable state and inflammatory responses simultaneously occur as evident by increase in D-Dimer and inflammatory biomarkers. While the overall thrombin generation potential in the plasma of these patients is decreased, thrombin is constantly generated in some of these patient leading to the formation of fibrin. The observed decrease in thrombin generation potential in these patients may be due to the consumption of prothrombin and other coagulation factors. Wide variations among these lymphoma groups are noted. Profiling of thrombin generation and inflammatory biomarkers may be helpful in thrombotic risk stratification of lymphoma patients and their anti-thrombotic management. Disclosures No relevant conflicts of interest to declare.

Microparticle-Associated Thrombin Generation and Procoagulant Activity Is Increased In Patients with Essential Thrombocythemia

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1985-1985
Author(s):  
Annamaria Leuzzi ◽  
Marina Marchetti ◽  
Marina Panova-Noeva ◽  
Carmen Julia Tartari ◽  
Laura Russo ◽  
...  
Keyword(s):  
Essential Thrombocythemia ◽  
Thrombin Generation ◽  
Lag Time ◽  
Procoagulant Activity ◽  
Jak2v617f Mutation ◽  
Wild Type ◽  
Thrombotic Risk ◽  
Time To Peak ◽  
Mutation Carriers ◽  
Free Plasma

Abstract Abstract 1985 Essential Thrombocythemia (ET) is a chronic myeloproliferative neoplasm characterized by an increased thrombotic risk. Numerous quantitative and qualitative abnormalities of platelets and leukocytes, arising from the clonal proliferation of hematopoietic progenitor cells, have been described as responsible of the thrombophilic state in this disease. Recently, a high number of plasma microparticles (MP) of different cellular origin has been described in ET patients. MP are a key component of the hemostatic response and have been found increased in diseases at high thrombotic risk. To explore the contribution of plasma MP to the hypercoagulable state of patients with ET, in this study we aimed to characterize the MP functional procoagulant features. We used two different methods: the calibrated automated thrombogram (CAT), to determine the MP-associated thrombin generation (TG), and the P-PPL/1 assay (Stago R&D) to measure the MP-associated procoagulant activity (PCA). Both assays were performed in platelet free plasma (P-FP) obtained from 69 ET patients (24M/45F; 33 carriers of JAK2V617F mutation) and 67 control subjects (32M/35F). In a subgroup of 23 ET patients and 23 controls MP-associated TG and PCA were also determined in MP-free plasma (MP-FP). PFP was obtained by two serial centrifugations (4,000 rpm for 15 min, then 11,000 rpm for 10 min) and MP-FP by re-centrifuging P-FP at 14,000 rpm for 30 min. MP were isolated from the pellet. For the TG assay, 80 ul of P-FP or MP-FP and 20 ul of buffer were mixed, and TG started by adding CaCl2. The results were expressed as lag-time, peak, area under the curve (ETP), and time-to-peak (ttPeak). For P-PPL/1 assay, 100 ul of P-FP or MP-FP were mixed with 50 ul of phospholipid-depleted plasma. Clotting was started by adding FXa and CaCl2 and results expressed in seconds. The results show that P-FP from ET patients generated significantly higher quantity of thrombin compared to controls, as demonstrated by the shorter lag-time (22.1±8.7 vs 29.5±14.7 min; p=0.004) and time to peak (26.6±8.4 vs 33.3±13.2 min; p=0.004) and the significantly greater peak (52.9±24.9 vs 38.24±20.9 nM; p=0.002) and ETP (765.6±206.4 vs 537.5±295 nM*min; p=0.001). Similarly, the MP-associated PCA was significantly increased in ET patients (79±11 sec) compared to controls (89±11 sec; p<0.05). This increase was due to the presence of MP, as no TG and little PCA was observed in MP-FP from both patients and controls. The addition of isolated MP to autologous MP-FP restored the TG and PCA of the samples to the original values of P-FP for both assays. TG was significantly (p<0.05) increased in the JAK2V617F mutation carriers (lag-time: 19.5±7.4 min; peak: 59.8±26.9 nM; ttpeak: 23.9± 4.9 min) compared to wild-type subjects (lag-time: 24.5±9.1 min, peak: 45.4±20.6 nM, ttpeak: 29.3±10.3 min), while no significant differences were found for PCA. Significant correlations were found between the PCA by PPL-assay and the different parameters of TG assay [lag-time (R2= 0.414), peak (R2= -0.542), ETP (R2= -0.514)]. In conclusion, our results show that MP-associated TG capacity, as well as PCA, are increased in plasma from ET patients. The highest MP-associated TG was found in JAK-2 mutation carriers, who also are at higher risk for thrombosis compared to wild-type subjects. Our data provide evidence for a contribution of MP to the thrombophilic state of these patients and suggest to test MP associated TG and PCA in prospective studies to predict thrombosis in ET patients. Disclosures: No relevant conflicts of interest to declare.

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