Thrombin Activatable Fibrinolysis Inhibitor Concentration Research Articles

Fibrinolytic Status and Risk of Death After Acute Pulmonary Embolism.

Acute pulmonary embolism (PE) is a heterogeneous disease process with variable presentation and outcomes. The endogenous fibrinolytic system is a complex framework of regulatory pathways that maintains homeostasis by dissolving overabundant thrombi. We sought to investigate phenotypic profiles of the endogenous fibrinolytic system among patients presenting with acute PE and their impact on mortality. We enrolled all consecutive patients with acute PE in our institutional Pulmonary Embolism Response Team registry. We collected blood samples at the time of PE diagnosis and analyzed concentrations of plasminogen activator inhibitor 1 (PAI-1), thrombin-activatable fibrinolysis inhibitor (TAFI), and alpha-2-antiplasmin (A2A). We assessed the association of concentration of fibrinolytic inhibitors and 1-year all-cause mortality and various echocardiographic markers of right ventricular (RV) dysfunction. There is significant variability of PAI-1, A2A, and TAFI concentrations across the spectrum of PE risk profiles with high PAI-1, low TAFI, and low A2A (herein referred to as a high-risk biomarker profile) correlating with worse PE severity. High-risk biomarker profile correlated with high-risk echocardiographic features of RV dysfunction, including increased RV/left ventricular (LV) ratio, low tricuspid annular plane systolic excursion, and low right ventricular outflow tract velocity time integral. Higher-risk biomarker profile was able to discriminate and independently identify patients at high risk of all-cause mortality (Group 2 HR 6 95% CI 1.3-27.8, Group 3 HR 12, 95% CI 1.7-86). Further studies are needed to assess the exact pathophysiological link between fibrinolytic status and poor outcome after acute PE and to ascertain the impact of anti-inhibitors of the fibrinolytic system on response to therapy and outcomes after acute PE.

A Case-Control Study of the Dose-Response Relationship Between Thrombin Activatable Fibrinolysis Inhibitor and Acute Myocardial Infarction.

BackgroundAcute myocardial infarction (AMI) is considered an acute coronary syndrome (ACS), which is caused by the death of myocardial cells after prolonged ischemia, and there is a high risk of sudden death during AMI. Therefore, the purpose of this study is to explore the relationship between thrombin activatable fibrinolysis inhibitor (TAFI) and AMI and provide evidence for their association and potentially the prevention of AMI.MethodsThere were 228 subjects included in this retrospective study, which included 78 AMI patients and 150 controls. The immune turbidimetry was used to measure TAFI concentration in the serum. Mann–Whitney U test was used to compare serum TAFI levels. The logistic regression analysis was used to construct a model of influencing factors of AMI. The dose-response relationship between serum TAFI level and AMI was explored by using the restricted cubic spline (RCS) functions combined with logistic regression analysis.ResultsThe serum TAFI levels of the AMI group were higher than the control group’s (P = 0.003). The risk of AMI in the high-TAFI level group was 2.24 times higher than the low-TAFI level group (P = 0.007) and it was 2.74 times higher after adjustment of other risk factors (P = 0.025). According to the dose-response curve, the risk of AMI increased significantly with an increase of serum TAFI concentration (P = 0.0387).ConclusionAcute myocardial infarction patients had higher serum TAFI levels, and TAFI was an independent risk factor for AMI patients. Serum TAFI levels demonstrated a dose- dependent response to the risk of AMI. Our study provides evidence that TAFI could be used for risk stratification of AMI patients.

Quantitation of thrombin-activatable fibrinolysis inhibitor in human plasma by isotope dilution mass spectrometry

Measurement of Thrombin-activatable fibrinolysis inhibitor (TAFI) in human plasma is dependent on reproducible assays. To date, standards for measuring TAFI are frequently calibrated relative to pooled normal human plasma and arbitrarily assigned a potency of 100% TAFI, despite variation in TAFI concentrations between plasma pools. Alternatively, TAFI calibrators can be assigned a value in SI units but the approach used for value assignment is not consistent and furthermore, if purified TAFI is used to determine TAFI concentration in plasma, may be adversely affected by matrix effects. A TAFI plasma standard in mass units with traceability to the SI unit of mass is desirable.We report here the establishment of a quantitative mass spectrometry method for TAFI in plasma. Traceability is obtained by reference to calibrators that consist of blank plasma spiked with a defined amount of purified TAFI, value assigned by amino acid analysis. The calibrators are run alongside the samples, using the same preparation steps and conditions; an acetonitrile assisted tryptic digestion and multi-dimensional liquid chromatography (LC) separation followed by MRM-MS analysis.We measured the TAFI quantitatively in human plasma with reproducibility, reliability and precision, and demonstrated the applicability of this approach for value assigning a common reference standard.

Comparison of the in-vivo effect of two tranexamic acid doses on fibrinolysis parameters in adults undergoing valvular cardiac surgery with cardiopulmonary bypass - a pilot investigation

BackgroundThe blood saving efficacy of TXA in cardiac surgery has been proved in several studies, but TXA dosing regimens were varied in those studies. Therefore, we performed this study to investigate if there is a dose dependent in-vivo effect of TXA on fibrinolysis parameters by measurement of fibrinolysis markers in adults undergoing cardiac surgery with CPB.MethodsA double-blind, randomized, controlled prospective trial was conducted from February 11, 2017 to May 05, 2017. Thirty patients undergoing cardiac valve surgery were identified and randomly divided into a placebo group, low-dose group and high-dose group by 1: 1: 1. Fibrinolysis parameters were measured by plasma levels of D-Dimers, plasminogen activator inhibitor-1 (PAI-1), thrombin activatable fibrinolysis inhibitor (TAFI), plasmin-antiplasmin complex (PAP), tissue plasminogen activator (tPA) and thrombomodulin (TM). Those proteins were measured at five different sample times: preoperatively before the TXA injection (T1), 5 min after the TXA bolus (T2), 5 min after the initiation of CPB (T3), 5 min before the end of CPB (T4) and 5 min after the protamine administration (T5). A Thrombelastography (TEG) and standard coagulation test were also performed.ResultsCompared with the control group, the level of the D-Dimers decreased in the low-dose and high-dose groups when the patients arrived at the ICU and on the first postoperative morning. Over time, the concentrations of PAI-1, TAFI, and TM, but not PAP and tPA, showed significant differences between the three groups (P < 0.05). Compared with the placebo group, the plasma concentrations of PAI-1 and TAFI decreased significantly at the T3 and T4 (P < 0.05); TAFI concentrations also decreased at the T5 in low-dose group (P < 0.05). Compared with the low-dose group, the concentration of TM increased significantly at the T4 in high-dose group.ConclusionsThe in-vivo effect of low dose TXA is equivalent to high dose TXA on fibrinolysis parameters in adults with a low bleeding risk undergoing valvular cardiac surgery with cardiopulmonary bypass, and a low dose TXA regimen might be equivalent to high dose TXA for those patients.Trial registrationChiCTR-IPR-17010303, Principal investigator: Zhen-feng ZHOU, Date of registration: January 1, 2017.

Levels of TAFI, TFPI and ADAMTS-13 in inflammatory bowel disease.

There is an increased tendency for thrombosis and thromboembolic complications in patients with inflammatory bowel disease (IBD). The aim of the present study was to determine the serum concentrations of thrombin-activatable fibrinolysis inhibitor (TAFI), tissue factor pathway inhibitor (TFPI) and a disintegrin and metalloproteinase with thrombospondin motif-13 (ADAMTS-13) in patients with IBD and to assess their possible role in the etiopathogenesis of the disease. Thirty-four patients with IBD (23 ulcerative colitis and 11 Crohn's disease) and 20 healthy controls were included in the present study. TAFI, TFPI, and ADAMTS-13 concentrations were determined by enzyme-linked immunosorbent assay. Mean TAFI, TFPI, and ADAMTS-13 concentrations in the patient group were 17.75 ng/ml, 72.10 ng/ml, and 14.90 U/l, respectively. In the control group, these values were 117.10 ng/ml, 300 ng/ml, and 191.55 U/l, respectively. TAFI, TFPI, and ADAMTS-13 values were significantly lower in the patient group than in the control group (all p<0.01). TAFI, TFPI, and ADAMTS-13 levels were significantly lower in the patient group. These findings indicate the presence of a clear, multifactorial imbalance in the coagulation-fibrinolytic system in the patient group. It is also possible that this imbalance in the coagulation and fibrinolytic system may play a role in the still unclear etiopathogenesis of the disease.

Reduced thrombin activatable fibrinolysis inhibitor and enhanced proinflammatory cytokines in acute coronary syndrome

ObjectiveA study was made of the changes in the serum levels of thrombin activatable fibrinolysis inhibitor (TAFI), proinflammatory cytokines and acute phase proteins in the acute stage of acute coronary syndrome (ACS), in order to explore the possibility of using TAFI as a biomarker for ACS risk assessment. MethodsA total of 211 patients with ACS were enrolled, and healthy subjects were used as controls. Blood samples were taken within 24h after admission. Serum TAFI levels were determined by immunoturbidimetry. Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA). Procalcitonin (PCT) and C-reactive protein (CRP) levels were measured by gold-immunochromatographic assay. ResultsSerum TAFI levels in ACS patients were significantly decreased versus the controls. The IL-1β, IL-6, TNF-α, PCT and CRP levels were markedly higher in the ACS patients than in the controls. Correlation analysis revealed a strong negative correlation between TAFI concentration and the IL-1β, IL-6, TNF-а, PCT and CRP levels in ACS patients and in controls. Multivariate logistic regression analysis suggested decreased serum TAFI to be an independent risk factor for ACS (OR 9.459; 95% CI 2.306–38.793; P=0.002). The area under the receiver operating characteristic (ROC) curve for TAFI was 0.872 (95% CI 0.787–0.909; P<0.001). The optimum TAFI cutoff point for the prediction of ACS was 24μg/ml, with a sensitivity of 75.83% and a specificity of 72.57%. ConclusionThese findings suggest that TAFI can be useful as a potential biomarker for ACS risk assessment.

Blood markers of fibrinolysis and endothelial activation in canine babesiosis

BackgroundCanine babesiosis is a tick-borne disease caused by hemoprotozoan parasites of the genus Babesia. The disease can be clinically classified into uncomplicated and complicated forms. The aim of this study was to assess the level of endothelial activation and alterations in the fibrinolytic pathway during canine babesiosis.ResultsBlood samples were collected on the day of admission and on the 6th day after treatment with imidocarb propionate, from 30 dogs of various breeds and of both sexes with naturally occurring babesiosis caused by B. canis. In this prospective study, plasminogen activity was assessed using a chromogenic assay, and concentrations of high mobility group box-1 protein (HMGB-1), intercellular adhesive molecule-1 (ICAM-1), vascular adhesive molecule-1 (VCAM-1), soluble urokinase receptor of plasminogen activator (suPAR), thrombin activatable fibrinolysis inhibitor (TAFI), soluble thrombomodulin (TM) and plasminogen activator inhibitor-1 (PAI-1) were determined using a canine specific ELISA. Concentrations of TM, HMGB-1, VCAM-1 and suPAR were increased in dogs with babesiosis at admission compared to healthy dogs. After treatment, concentrations of TM were lower in infected dogs compared to healthy dogs. Dogs with babesiosis also had increased concentrations of TM, ICAM-1 and HMGB-1 and decreased plasminogen and PAI-1 at presentation compared to day 6 after treatment. Dogs with complicated babesiosis had higher concentrations of TM, HMGB1 and TAFI at admission compared to the 6th day.ConclusionsBiomarkers of endothelial activation and fibrinolysis were altered in dogs with babesiosis. Further studies into their usefulness as biomarkers of disease severity or prognosis is warranted.

Reduced thrombin activatable fibrinolysis inhibitor and enhanced proinflammatory cytokines in acute coronary syndrome

ObjectiveA study was made of the changes in the serum levels of thrombin activatable fibrinolysis inhibitor (TAFI), proinflammatory cytokines and acute phase proteins in the acute stage of acute coronary syndrome (ACS), in order to explore the possibility of using TAFI as a biomarker for ACS risk assessment. MethodsA total of 211 patients with ACS were enrolled, and healthy subjects were used as controls. Blood samples were taken within 24h after admission. Serum TAFI levels were determined by immunoturbidimetry. Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA). Procalcitonin (PCT) and C-reactive protein (CRP) levels were measured by gold-immunochromatographic assay. ResultsSerum TAFI levels in ACS patients were significantly decreased versus the controls. The IL-1β, IL-6, TNF-α, PCT and CRP levels were markedly higher in the ACS patients than in the controls. Correlation analysis revealed a strong negative correlation between TAFI concentration and the IL-1β, IL-6, TNF-а, PCT and CRP levels in ACS patients and in controls. Multivariate logistic regression analysis suggested decreased serum TAFI to be an independent risk factor for ACS (OR 9.459; 95% CI 2.306–38.793; P=0.002). The area under the receiver operating characteristic (ROC) curve for TAFI was 0.872 (95% CI 0.787–0.909; P<0.001). The optimum TAFI cutoff point for the prediction of ACS was 24μg/ml, with a sensitivity of 75.83% and a specificity of 72.57%. ConclusionThese findings suggest that TAFI can be useful as a potential biomarker for ACS risk assessment.

The Occurrence of Thrombosis in Inflammatory Bowel Disease Is Reflected in the Clot Lysis Profile.

The occurrence of thromboembolic events (TE) is an important extraintestinal manifestation in patients with inflammatory bowel disease (IBD). The aim of this study was to compare fibrinolysis and clot lysis parameters between (1) patients with IBD and healthy controls and (2) patients with IBD with TE (IBD + TE) and without TE (IBD - TE). One hundred thirteen healthy controls and 202 patients with IBD, of which 84 patients with IBD + TE and 118 patients with IBD - TE, were included in this case-control study. Three clot lysis parameters (area under the curve, 50% clot lysis time, and amplitude) were determined using a clot lysis assay. Plasminogen activator inhibitor 1 (PAI-1) and thrombin activatable fibrinolysis inhibitor concentrations were determined by enzyme-linked immunosorbent assay. PAI-1 antigen, active PAI-1, and intact thrombin activatable fibrinolysis inhibitor concentrations, as well as 50% clot lysis time and area under the curve, were significantly associated with the presence of IBD (all P < 0.05). The median time between TE and plasma collection was 5.0 (1.8-11.0) years. Comparing IBD + TE versus IBD - TE, active to total PAI-1 ratio (0.36 [0.24-0.61] versus 0.24 [0.13-0.40]), area under the curve (31 [24-49] versus 22 [13-31]), 50% clot lysis time (110 [64-132] versus 95 [70-126] minutes), and amplitude (0.295 [0.222-0.436] versus 0.241 [0.168-0.308]) were significantly higher in IBD + TE (all P <0.05) and remained higher after adjustment for age, gender, C-reactive protein, type of disease, presence of comorbidities, and disease activity. Patients with IBD have an altered clot lysis profile compared with healthy controls. Clot lysis parameters differ significantly between patients with IBD with and without a history of TE and should be included in the risk assessment.

Active PAI-1 as marker for venous thromboembolism: Case–control study using a comprehensive panel of PAI-1 and TAFI assays

BackgroundBoth activated Thrombin Activatable Fibrinolysis Inhibitor (TAFI) and active Plasminogen Activator Inhibitor-1 (PAI-1) attenuate fibrinolysis and may therefore contribute to the pathophysiology of Venous ThromboEmbolism (VTE). Whether increased TAFI and/or PAI-1 concentrations are associated with VTE is unclear. ObjectiveTo study an association of impaired fibrinolysis and VTE using a comprehensive panel of in-house developed assays measuring intact TAFI, activation peptide of TAFI (AP-TAFI), PAI-1 antigen, endogenous PAI-1:t-PA complex (PAI-1:t-PA) and active PAI-1 levels in 102 VTE patients and in 113 healthy controls (HC). ResultsActive PAI-1 was significantly higher in VTE patients compared to HC (20.9 [9.6-37.8] ng/ml vs. 6.2 [3.5-9.7] ng/ml, respectively). Active PAI-1 was the best discriminator with an area under the ROC curve and 95% confidence interval (AUROC [95%CI]) of 0.84 [0.79-0.90] compared to 0.75 [0.68-0.72] for PAI-1:t-PA, 0.65 [0.58-0.73] for PAI-1 antigen, 0.62 [0.54-0.69] for AP-TAFI and 0.51 [0.44-0.59] for intact TAFI. Using ROC analysis, we defined an optimal cut-off of 12.8ng/ml for active PAI-1, with corresponding sensitivity of 71 [61–79] % and specificity of 89 [82–94] %. A lack of association with the time between VTE event and sample collection or with the intake of anticoagulant treatment suggests that active PAI-1 levels are sustainable high in VTE patients. ConclusionsThis case–control study emphasizes the clinical importance of measuring active PAI-1 instead of PAI-1 antigen and identifies active PAI-1 as a potential marker of VTE. Prognostic studies will need to address the clinical significance of active PAI-1 as biomarker.

Regulation of the gene encoding human thrombin-activatable fibrinolysis inhibitor by estrogen and progesterone

Thrombin-activatable fibrinolysis inhibitor (TAFI) is a pro-carboxypeptidase B-like pro-enzyme that upon activation attenuates fibrinolysis and inflammatory processes. There is a large inter-individual variability in plasma TAFI levels within the population which is primarily due to epigenetic factors. Novel associations between plasma TAFI levels and sex steroids have triggered interest in determining the role of TAFI as a mediator of the cardioprotective effects of estrogens and progestins, or as a mediator of the increased thrombotic risk that accompanies use of oral contraceptives or hormone replacement therapy (HRT). In this study, we measured the effect of sex steroids on hepatic expression of CPB2, the human gene encoding TAFI. Using human hepatocellular carcinoma cells cultured in the presence of progesterone and 17β-estradiol, we demonstrated that the level of TAFI protein is decreased by those sex steroids. These changes in protein expression were paralleled by decreases in CPB2 mRNA abundance and promoter activity. We did not find evidence of estrogen or progesterone receptor binding sites in the CPB2 promoter region, suggesting that the genomic effects of progesterone and 17β-estradiol are mediated indirectly, without receptor binding to the CPB2 promoter. Indeed, we found that the effect of estrogen was independent of the estrogen receptor and was mediated through a novel signaling pathway dependent on phosphatidylinositol 3-kinase and Akt that did not involve GPR30. Our findings provide the molecular explanation for the ability of female sex steroids to decrease plasma TAFI concentrations.

Thrombin Activatable Fibrinolysis Inhibitor (TAFI) Levels in Hypertensive Patients and a Comparison of the Effects of Amlodipine and Ramipril on TAFI Levels

Hypertension is associated with fibrinolysis abnormality. Thrombin-activatable fibrinolysis inhibitor (TAFI) is a novel molecule-linking coagulation and fibrinolysis. The aim of this study was to investigate the levels of TAFI in primary hypertensive patients and to compare the effects of amlodipine and ramipril on TAFI levels. The study was performed with 58 hypertensive subjects and 27 healthy volunteers. Biochemical and hematological parameters and TAFI levels were measured at baseline and after 1-month follow-up. TAFI concentrations increased in hypertensive patients compared with the controls (P = .030). Additionally, TAFI levels decreased with blood pressure control at 1-month follow-up (P = .026). There was no significant difference between TAFI levels in the amlodipine and ramipril groups at baseline. However, after 1-month follow-up, TAFI levels were decreased in the amlodipine group (P = .037) but not in the ramipril group. Our study is the first in the literature to determine increased TAFI levels in primary hypertension patients. In addition, we determined a decrease in TAFI levels in the amlodipine group after 1 month, but none in the ramipril group.

Flexibility of the Thrombin-activatable Fibrinolysis Inhibitor Pro-domain Enables Productive Binding of Protein Substrates

We have previously reported that thrombin-activatable fibrinolysis inhibitor (TAFI) exhibits intrinsic proteolytic activity toward large peptides. The structural basis for this observation was clarified by the crystal structures of human and bovine TAFI. These structures evinced a significant rotation of the pro-domain away from the catalytic moiety when compared with other pro-carboxypeptidases, thus enabling access of large peptide substrates to the active site cleft. Here, we further investigated the flexible nature of the pro-domain and demonstrated that TAFI forms productive complexes with protein carboxypeptidase inhibitors from potato, leech, and tick (PCI, LCI, and TCI, respectively). We determined the crystal structure of the bovine TAFI-TCI complex, revealing that the pro-domain was completely displaced from the position observed in the TAFI structure. It protruded into the bulk solvent and was disordered, whereas TCI occupied the position previously held by the pro-domain. The authentic nature of the presently studied TAFI-inhibitor complexes was supported by the trimming of the C-terminal residues from the three inhibitors upon complex formation. This finding suggests that the inhibitors interact with the active site of TAFI in a substrate-like manner. Taken together, these data show for the first time that TAFI is able to form a bona fide complex with protein carboxypeptidase inhibitors. This underlines the unusually flexible nature of the pro-domain and implies a possible mechanism for regulation of TAFI intrinsic proteolytic activity in vivo.

Secretion and antifibrinolytic function of thrombin-activatable fibrinolysis inhibitor from human platelets.

The thrombin-activatable fibrinolysis inhibitor (TAFI) is a zymogen first characterized in human plasma that is activated through proteolytic cleavage by thrombin, thrombin in complex with thrombomodulin, or plasmin. Active TAFI attenuates fibrinolysis by removing C-terminal lysine residues from partially degraded fibrin, thereby inhibiting a potent positive feedback loop in the fibrinolytic cascade. The existence of a separate pool of TAFI within platelets has been described. We aimed to confirm the presence of TAFI in the medium of washed, thrombin-stimulated platelets and to evaluate the characteristics of platelet TAFI by western blot analysis and with a quantitative assay for activated TAFI. We also assessed the ability of platelet TAFI to inhibit fibrinolysis in vitro, using a platelet-rich thrombus lysis assay. Our data are consistent with the presence of TAFI in the α-granules of resting platelets. In contrast to previous reports, platelet TAFI is very similar in electrophoretic mobility to plasma-derived TAFI. We also show, for the first time, that platelet-derived TAFI is capable of attenuating platelet-rich thrombus lysis in vitro independently of plasma TAFI. Moreover, we demonstrate additive effects on thrombolysis of platelet-derived TAFI and TAFI present in plasma. Taken together, these observations indicate that the secretion of platelet-derived TAFI can augment the concentrations of TAFI already present in plasma to enhance attenuation of the fibrinolytic cascade. This could be significant in regions of vascular damage or pathologic thrombosis, where activated platelets are known to accumulate.

Valor pronóstico de la concentración del inhibidor de la fibrinolisis activable por trombina y del polimorfismo C1040T en el infarto agudo de miocardio tratado con fibrinolisis

To analyze the prognostic value of thrombin activatable fibrinolysis inhibitor (TAFI) and C1040T polymorphism in acute myocardial infarction treated with fibrinolysis. To analyze C1040T polymorphism influence on its plasma level. An observational, prospective study performed from November 2003 to November 2005 and with a 3 month follow-up. Intensive Medicine Service from a university-affiliated teaching hospital. A total of 53 patients with acute myocardial infarction with persistent ST segment elevation treated with the same fibrinolytic therapy. A control group of 53 biologically similar subjects was included. None. Baseline characteristics; frequency of wild-type genotype (Thr325Thr) and of those corresponding to the mutation (Thr325LLe and LLe325lle), TAFI levels at 6 h, 34 h and 3 months post-fibrinolysis; ejection fraction; Killip-Kimball; reperfusion; ischemic recurrence; death. No relationship was found between biological features and TAFI concentration. The latter was significantly higher in infarct patients (p<0.01) and in the mutation group (p<0.01). The homozygotic mutation (Ile325Ile) was significantly higher in infarct patients (p<0.01). Reperfusion was significantly associated with lower body mass index (p=0.02. OR 0.22. 95% CI), ejection fraction (p=0.004. OR 0.91. 95% CI), triglyceride level (p=0.01. OR 1.02. 95% CI) and cholesterol levels (p=0.001. OR=0.84. 95% CI). Mutation was associated to a significant fall in post-fibrinolysis concentration TAFI antigen and functional TAFI (p=0.01) and (p=0.02), and lower frequency of reperfusion. Reperfusion was associated with a significant post-fibrinolysis reduction in the level of TAFI antigen (p=0.02). Recurrence was associated to a significantly higher post-fibrinolysis level (p=0.05. OR=0.84. 95% CI). This was more frequent in mutation. Post-fibrinolysis TAFI antigen concentration was significantly lower in non-recurrence patients (p=0.028. OR=1.03. 95% CI). A higher concentration of TAFI is associated to a worse prognosis in reperfusion and recurrence in acute myocardial infarction treated with fibrinolysis. Homozygotic mutation was more frequent in myocardial infarction patients. Wild genotype is associated to a better prognosis. Mutation is associated to a higher expression of TAFI.

Assessment of thrombin-activatable fibrinolysis inhibitor (TAFI) activation in acquired hemostatic dysfunction: a diagnostic challenge

Human thrombin activatable fibrinolysis inhibitor (TAFI) is a carboxypeptidase zymogen that was first described in 1995 [1]. It had been previously reported under different names such as carboxypeptidase basic (B), unstable (U), and arginine (R) [2]. The mature TAFI protein contains 401 amino acids, and after glycosylation, the molecular weight becomes 60 kDa. TAFI in its plasma zymogen form can be activated by thrombin, the thrombin-thrombomodulin complex, or plasmin. The activated TAFI (TAFIa) decays spontaneously into the inactive form of TAFI (TAFIai) through a temperature-dependent conformational change. TAFIa plays a role in the inhibition of fibrinolysis by cleaving the carboxyl-terminal lysine residues from partially degraded fibrin. In the coagulation cascade, the generated thrombin can activate TAFI, and the resulting TAFIa inhibits fibrinolysis, finally resulting in the formation of more stable clots [3]. Therefore, a deficiency of the coagulation factors might reduce the TAFIa levels via reduced thrombin generation, resulting in the formation of an unstable clot. In an animal study, TAFI was activated during a thrombotic event, and the inhibition of TAFIa activity enhanced the efficacy of thrombolytic therapy, thereby confirming its role as an inhibitor of fibrinolysis [4-6]. TAFIa also plays a role in the modulation of inflammation through down-regulation of pericellular plasminogen activation and inactivation of the inflammatory peptides bradykinin and anaphylatoxins C3a and C5a. In mice with Escherichia coli-induced sepsis, TAFI deficiency prevented liver damage via inactivation of C3a and C5a [7]. The plasma concentration of TAFI appears to be under the control of genetic factors and non-genetic factors such as sex hormones, cytokines, and lipids [2]. Several diseases, including diabetes, kidney transplantation, hypertension, nephritic syndrome, insulin resistance, obesity, and inflammatory bowel disease, have been shown to be positively associated with plasma TAFI levels [2]. Because TAFI is expressed in adipocytes, an increased TAFI level can be expected in metabolic disturbance. The TAFI levels were also elevated in thrombotic diseases. In keeping with the role of TAFI as an antifibrinolytic factor, high plasma level of TAFI is a risk factor for thrombotic diseases. Meanwhile, TAFI levels are reduced in acquired hemostatic dysfunctions, such as post-partum hemorrhage and sepsis accompanying disseminated intravascular coagulation [3], because the enhanced generation of thrombin or plasmin may result in enhanced TAFI activation and consequent consumption of the TAFI zymogen. Many different methods for measuring TAFI by using ELISA, enzymological methods, or functional methods have been described. All the methods have their inherent advantages and disadvantages, and to date, there is no reference standard for TAFI. Most studies have used the total antigen level of TAFI, but this has yielded ambiguous results, because of the existence of different isoforms and various degrees of activation. Ceres et al. [8] have reported the use of an ELISA that specifically detects TAFI activation. Since substantial TAFI activation occurs in acquired hemostatic dysfunction, ELISA that measures the extent of TAFI activation is potentially useful. In this issue of the Korean Journal of Hematology, Park et al. [9] reported high levels of TAFI activation in sepsis patients by measuring TAFIa/ai levels using an ELISA that they developed in-house. In the in-house ELISA, potato tuber carboxypeptidase inhibitor was used as a capture agent that specifically captured the activated and inactivated TAFI isoforms and did not capture the intact TAFI zymogen. Although only a few septic patients (n=25) were enrolled in the study, the activation status of TAFI could clearly differentiate septic patients from healthy controls. This result is in agreement with the results of a recent study [10] that reported a significant elevation of TAFI activation in meningococcal infection. Since decreases in total TAFI level are associated with increases of TAFI activation level, simultaneous determination of total TAFI and TAFI activation marker is required to interpret the role of TAFI in acquired hemostatic dysfunctions including sepsis. Although the principal physiologic role of TAFI is still a matter of debate, TAFI is reported to be an important protein in the fibrinolytic system. The advent of more accurate assays for measuring plasma TAFI levels may help validate the mechanism underlying TAFI activation in acquired hemostatic dysfunction. This information may help elucidate the contribution of the TAFI pathway to acquired hemostatic dysfunction and will form the basis of potential therapeutic strategies to manipulate the TAFI pathway for acquired hemostatic dysfunction.

Elevated levels of activated and inactivated thrombin-activatable fibrinolysis inhibitor in patients with sepsis

BackgroundIn sepsis, large scale inflammatory responses can cause extensive collateral damage to the vasculature, because both coagulation and fibrinolysis are activated unevenly. Thrombin-activatable fibrinolysis inhibitor (TAFI) plays a role in modulating fibrinolysis. Since TAFI can be activated by both thrombin and plasmin, it is thought to be affected in sepsis. Hence, activated and inactivated TAFI (TAFIa/ai) may be used to monitor changes in sepsis.MethodsTAFIa/ai-specific in-house ELISA can detect only the TAFIa/ai form, because the ELISA capture agent is potato tuber carboxypeptidase inhibitor (PTCI), which has selective affinity towards only the TAFIa and TAFIai isoforms. TAFIa/ai levels in plasma from 25 patients with sepsis and 19 healthy volunteers were quantitated with the in-house ELISA.ResultsWe observed increased TAFIa/ai levels in samples from patients with sepsis (48.7±9.3 ng/mL) than in samples from healthy individuals (10.5±5.9 ng/mL). In contrast, no difference in total TAFI concentration was obtained between sepsis patients and healthy controls. The results suggest that TAFI zymogen was activated and that TAFIa/ai accumulated in sepsis.ConclusionThe detection of TAFIa/ai in plasma could provide a useful and simple diagnostic tool for sepsis. Uneven activation of both coagulation and fibrinolysis in sepsis could be caused by the activation of TAFI zymogen and elevation of TAFIa/ai. TAFIa/ai could be a novel marker to monitor sepsis and other blood-related disturbances.

Presence of Thrombin‐Activatable Fibrinolysis Inhibitor in Helicobacter pylori‐Associated Gastroduodenal Disease

Thrombin-activatable fibrinolysis inhibitor (TAFI) plays a role in the regulation of coagulation and inflammation. In addition to inhibiting the fibrinolytic system, TAFI may also regulate the bradykinin and complement systems. We hypothesized that TAFI also plays a role in defense mechanisms of the gastric mucosa during Helicobacter pylori infection. This study comprised 65 patients with gastroduodenal disorders: 41 patients with H. pylori infection, 13 without, and 11 patients with cured H. pylori infection. The gastric intramucosal concentrations of TAFI were measured by enzyme immunoassay. The gastric levels of TAFI and plasminogen activator inhibitor-1 were significantly increased in patients with H. pylori compared to those without infection or cured H. pylori. The presence of TAFI was detected in gastric mucosal epithelial cells. The concentration of TAFI was correlated with the degree of gastric mucosal atrophy, inflammation, and disease activity. These results show that TAFI is present in the gastric mucosa and that it may play a role in the pathogenesis of H. pylori infection-associated gastroduodenal disorders.

Fenofibrate improves endothelial function and decreases thrombin-activatable fibrinolysis inhibitor concentration in metabolic syndrome

Procoagulant state, inflammation, and endothelial dysfunction have been documented in metabolic syndrome. Endothelial dysfunction is a strong predictor of cardiovascular events. Studies on the association of thrombin-activatable fibrinolysis inhibitor and thrombosis are still controversial, but substantial evidence suggests that increased thrombin-activatable fibrinolysis inhibitor or thrombin-activatable fibrinolysis inhibits or protects against arterial thrombosis. This study aimed to assess concomitantly the effects of fenofibrate therapy on thrombin-activatable fibrinolysis inhibitor concentrations and endothelial functions in patients with metabolic syndrome. Twenty-five patients (16 women; mean age 50.4 +/- 7.0) were enrolled in the study. Plasma thrombin-activatable fibrinolysis inhibitor, C-reactive protein, and fibrinogen levels were measured before fenofibrate administration and after 8 weeks of fenofibrate treatment. Endothelial function was assessed by endothelial-dependent flow-mediated dilatation from brachial artery. Pretreatment (baseline) thrombin-activatable fibrinolysis inhibitor level was 52.3 (1.2-119.7) decreasing to 7.7 (0.9-51.2; P < 0.001) after 8 weeks of fibrate treatment. Endothelial functions, which were measured with flow-mediated dilatation, were significantly improved after treatment (mean flow-mediated dilatation was 6.76 +/- 2.21 at baseline and 10.66 +/- 1.17% after 8 week of fenofibrate treatment, P < 0.001). Fenofibrate decreases thrombin-activatable fibrinolysis inhibitor levels and improves endothelial function in metabolic syndrome and, thus, suggests a potential for protection against cardiovascular effects. Further studies are warranted to confirm the effects of fibrates on thrombin-activatable fibrinolysis inhibitor and for conclusive evidence on the association between thrombin-activatable fibrinolysis inhibitor and thrombosis.

Thrombin activatable fibrinolysis inhibitor and other hemostatic parameters in patients with essential arterial hypertension

Hypertension is associated with hemostatic abnormalities and endothelial dysfunction. thrombin activatable fibrinolysis inhibitor (TAFI) is a glycoprotein linking coagulation and fibrinolysis. Objectives. We evaluated TAFI concentrations in patients with essential hypertension in relation to blood pressure. Additionally, we studied TAFI activator, thrombin activity (thrombin-antithrombin complexes--TAT, prothrombin fragments F1 + 2), thrombomodulin (TM)--a marker of endothelial cell injury, degree of plasmin generation (plasmin-antiplasmin complexes [PAP]), other markers of endothelial cell injury--von Willebrand factor (vWF). Seventy-two patients with essential hypertension (27 untreated, 13 treated with enalapril (angiotensin-converting enzyme inhibitor [ACEI]), 32 with beta-blocker, betaxolol). In every hypertensive patients ambulatory blood pressure measurements and echocardiography were performed. All hypertensive patients did not differ with respect to age, creatinine, fibrinogen, D-dimers. In ACEI-treated patients a significantly higher TAFI concentration was observed when compared to beta-blocker-treated patients. In beta-blocker-treated patients both diastolic and systolic blood pressure were lower than in ACEI treated patients as well as ejection fraction, while serum triglycerides were higher. Diastolic blood pressure correlated significantly with TAFI concentrations in untreated patients (r = 0.27, p < 0.05), and in beta-blocker-treated patients (r = 0.25, p = 0.05), TAFI activity was inversely associated with interventricular septal diameter (r = -0.75, p < 0.01) in patients treated with ACEI. Elevated TAFI concentrations and enhanced thrombin generation in hypertensive patients may contribute to atherosclerosis progression in this population. Differences in the studied parameters may be due to a small sample size, monotherapy and potential effects of antihypertensive drugs on glicemia, ejection fraction and triglycerides.