Figure 2 - available via license: Creative Commons Attribution 4.0 International
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A clot waveform analysis for thrombin time. (a) Calibration plasma; (b) FII-deficient plasma; (c) FV-deficient plasma; (d) FVII-deficient plasma; (e) FVIII-deficient plasma; (f) FIX-deficient plasma; (g) FX-deficient plasma; (h) FXIdeficient plasma; (i) FXII-deficient plasma; (j) FXIII-deficient plasma; thrombin 0.5 IU/mL. Navy line, fibrin formation curve; red line, 1st derivative curve (velocity); light blue, 2nd derivative curve (acceleration).
Source publication
Object:
Although thrombin burst has attracted attention as a physiological coagulation mechanism, clinical evidence from a routine assay for it is scarce. This mechanism was therefore evaluated by a clot waveform analysis (CWA) to assess the thrombin time (TT).
Material and methods:
The TT with a low concentration of thrombin was evaluated using...
Context in source publication
Context 1
... total of 5 IU/mL of thrombin showed similar CWA-TT between calibration and FVIII-deficient plasma samples. The heights of the 2nd DP, 1st DP, and FFC in FII-, FV-, FVII-, FVIII-, FIX-, FX-, FXI-, and FXII-deficient plasma using the CWA-TT with 0.5 IU thrombin were significantly lower than those in calibration plasma and FXIII-deficient plasma using the CWA-TT with 0.5 IU thrombin ( Figure 2). In particular, the heights of the 2nd DP, 1st DP, and FFC in FVIIIdeficient plasma were extremely low. ...
Citations
... reported to be caused by a thrombin burst. [53][54][55][56] Although the mechanism underlying the prolonged clotting time in patients with HCC may involve hepatic dysfunction, the concomitance of both prolonged peak times and increased peak heights cannot be sufficiently understood. These findings in the CWA of prolonged peak times and increased peak heights indicate that a strong coagulation process might continue for a long time, suggesting that a thrombotic risk may exist in patients with HCC. ...
Background: Although hepatocellular carcinoma (HCC) is frequently associated with thrombosis, it is also associated with liver cirrhosis (LC) which causes hemostatic abnormalities. Therefore, hemostatic abnormalities in patients with HCC were examined using a clot waveform analysis (CWA). Methods: Hemostatic abnormalities in 88 samples from HCC patients, 48 samples from LC patients and 153 samples from patients with chronic liver diseases (CH) were examined using a CWA-activated partial thromboplastin time (APTT) and small amount of tissue factor induced FIX activation (sTF/FIXa) assay. Results: There were no significant differences in the peak time on CWA-APTT among HCC, LC, and CH, and the peak heights of CWA-APTT were significantly higher in HCC and CH than in HVs and LC. The peak heights of the CWA-sTF/FIXa were significantly higher in HCC than in LC. The peak times of the CWA-APTT were significantly longer in stages B, C, and D than in stage A or cases of response. In the receiver operating characteristic (ROC) curve, the fibrin formation height (FFH) of the CWA-APTT and CWA-sTF/FIXa showed the highest diagnostic ability for HCC and LC, respectively. Thrombosis was observed in 13 HCC patients, and arterial thrombosis and portal vein thrombosis were frequently associated with HCC without LC and HCC with LC, respectively. In ROC, the peak time×peak height of the first derivative on the CWA-sTF/FIXa showed the highest diagnostic ability for thrombosis. Conclusion: The CWA-APTT and CWA-sTF/FIXa can increase the evaluability of HCC including the association with LC and thrombotic complications.
... Elevated PT and APTT values signify prolonged clotting time, impaired coagulation function, and increased bleeding risk [17,18]. TT evaluates coagulation function by examining plasma fibrinogen's capacity to form fibrin polymers [19]. Prolonged TT indicates potential dysfunction in fibrinogen-to-fibrin conversion [20]. ...
... FIB is the substrate of the process, so its level affects the length of TT. 30,31 Based on the activation of FVIII, TT can reflect a thrombin burst, which was synergistic with the generation of FIXa caused by the process that APTT activated the intrinsic pathway, thus contributing to the conversion of FIB to fibrin. 31,32 In our study, the results suggested the decrease of TT with accompanying by the increase of FIB. The shorter TT in the adenomyosis patients than in the patients with uterine leiomyoma or the control group may be due to the higher level of FIB in the adenomyosis. ...
Objective
Adenomyosis patients are in a hypercoagulable state, and studies have shown that carbohydrate antigen125 (CA125) may relate to the hypercoagulability and thrombosis of patients with adenomyosis, but there is still a lack of clarity regarding the changes in CA125-related coagulation indicators. This study was to explore the changes and influencing factors of CA125-related coagulation parameters in patients with adenomyosis.
Methods
Retrospective observational study conducted on 200 patients with adenomyosis (AM group), 240 patients with uterine leiomyoma (LM group) and 81 patients with cervical intraepithelial neoplasia (CIN)-III (control group), of which the coagulation parameters were detected by clinical blood sample collection and statistical method analysis and informed consent was obtained.
Results
The level of CA125 in the AM group was significantly higher than that in the LM group and control group. However, thrombin time (TT) shortened in the AM group when compared with the LM and control group. Activated partial thromboplastin time (APTT) in the AM group was shorter than in the control group. Multivariate logistic regression analysis found that adenomyosis was associated with CA125 level (OR=323.860, 95% CI 90.424–1159.924, P<0.001), APTT (OR=1.295, 95% CI 1.050–1.598, P=0.016), TT (OR=0.642, 95% CI 0.439–0.938, P=0.022), menorrhagia (OR=7.363, 95% CI 2.544–21.315, P<0.001), dysmenorrhea (OR=22.590, 95% CI 8.185–62.347, P<0.001). Correlation analysis revealed that APTT (r= −0.207) and TT (r = −0.174) were negatively correlated with the level of CA125.
Conclusion
The shortening of CA125-related APTT and TT indicates that it is meaningful to detect coagulation parameters of patients with elevated CA125 levels early, dysmenorrhea and menorrhagia, and maybe further discover the hypercoagulability and prevent the occurrence of thrombus in adenomyosis.
... Furthermore, the use of a small scale of TF-induced FIX activation (sTF/FIXa) (CWA-sTF/ FIXa) assay can evaluate hemostatic abnormalities including platelet abnormalities [17]. Hypercoagulability is considered to be caused by a thrombin burst [20,21] which is evaluated by thrombin time using a small amount of thrombin with a CWA (CWA-TT) [22]. ...
... The CWA-TT was measured using 0.5 IU thrombin (Thrombin 500 units Mochida Pharmaceutical CO., LTD, Tokyo, Japan) with an ACL-TOP ® system (Instrumentation Laboratory) [17]. Three types of curves are shown on this system monitor [22]. The 1 st DP showed two peaks and the second peak height of the 1 ST DP (1 st DPH-2) reflected the thrombin burst. ...
... Markedly high peak heights of the sTF/FIXa using small amount of TF suggest that intrinsic TF released from cancer cells exists in plasma. Thrombin causes an elevation in FVIII activity and increases the activation cycle from thrombin to FXIa, called the thrombin burst [22]. Elevated peak heights, especially the second peak of the 1 st DPH (1 st DPH-2) of CWA-TT suggest that the thrombin burst worsens hypercoagulability in cancer patients including those with pancreatic cancer. ...
Background: Cancer, especially pancreatic cancer, is frequently associated with thrombosis which is one of the causes of poor outcomes; moreover hypercoagulability can be present in cancer patients. Hypercoagulability is considered to be caused by a thrombin burst. Methods: Activated Partial Tthromboplastin Time (APTT), small amount of tissue factor induced FIX activation assay (sTF/FIXa) and Thrombin Time (TT) assessment using Clot Waveform Analysis (CWA) were performed in 138 patients with malignant neoplasms, including pancreatic cancer. Results: The first derivative peak (1st DP) time (1st DPT), 1st DP height (1st DPH) and 1st DPH/1st DPT ratio were increased in a clotting-factor-FVIII-dependent manner. Thrombosis was frequently associated with pancreatic cancer and was observed in the early stage. CWA-APTT and CWA-sTF/FIXa indicated that the peak times and heights were markedly longer and higher, respectively, in cancer patients, especially pancreas cancer patients, than in patients without cancer. The 1st DPH/1st DPT ratios of CWA-sTF/FIXa were significantly high in patients with pancreas cancer (median value 1.5). CWA-TT showed that the peak times were significantly shorter in cancer patients than in healthy volunteers and that the peak heights were significantly higher in cancer than in benign pancreas diseases. The cutoff value of the 1st DPH/ 1st DPT of sTF/FIXa for cancer patients with thrombosis vs. all patients without cancer was 1.3. Conclusions: Cancer patients, including those with pancreatic cancer were frequently associated with thrombosis due to hypercoagulability caused by thrombin burst detected by CWA. A high 1st DPH/1st DPT ratio of sTF/FIXa may suggest an association with cancer or thrombosis.
... Although PT was the most important indicator for detecting the coagulation status of patients, it was influenced by various factors such as liver synthesis function and inflammatory factors [42]. TT was the time at which fibrinogen was converted into fibrin after the addition of thrombin, and the prolongation of TT to a certain extent reflected the level and state of fibrinogen [43]. The decrease in fibrinogen levels also indicated liver synthesis dysfunction, reflecting long-term damage to liver function [44]. ...
Objective
The predictive biomarkers of immune checkpoint inhibitors (ICIs) in hepatocellular carcinoma (HCC) still need to be further explored. This study aims to establish a new immune prognosis biomarker to predict the clinical outcomes of hepatocellular carcinoma patients receiving immune checkpoint inhibitors.
Methods
The subjects of this study were 151 HCC patients receiving ICIs at Harbin Medical University Cancer Hospital from January 2018 to December 2021. This study collected a wide range of blood parameters from patients before treatment and used Cox’s regression analysis to identify independent prognostic factors in blood parameters, as well as their β coefficient. The hepatocellular carcinoma immune prognosis score (HCIPS) was established through Lasso regression analysis and COX multivariate analysis. The cut-off value of HCIPS was calculated from the receiver operating characteristic (ROC) curve. Finally, the prognostic value of HCIPS was validated through survival analysis, stratified analyses, and nomograms.
Results
HCIPS was composed of albumin (ALB) and thrombin time (TT), with a cut-off value of 0.64. There were 56 patients with HCIPS < 0.64 and 95 patients with HCIPS ≥ 0.64, patients with low HCIPS were significantly related to shorter progression-free survival (PFS) (13.10 months vs. 1.63 months, P < 0.001) and overall survival (OS) (14.83 months vs. 25.43 months, P < 0.001). HCIPS has also been found to be an independent prognostic factor in this study. In addition, the stratified analysis found a significant correlation between low HCIPS and shorter OS in patients with tumor size ≥ 5 cm ( P of interaction = 0.032). The C-index and 95% CI of the nomograms for PFS and OS were 0.730 (0.680–0.779) and 0.758 (0.711–0.804), respectively.
Conclusions
As a new score established based on HCC patients receiving ICIs, HCIPS was significantly correlated with clinical outcomes in patients with ICIs and might serve as a new biomarker to predict HCC patients who cloud benefit from ICIs.
... However, few reports have described the relationship between FVIII activity, which is assessed using the peak time and height of CWA-APTT, including a small amount of tissue factor-induced activated FIX (sTF/FIXa) assay [15]. A small amount of thrombin reflects the thrombin burst, including the activation of FXI, FVIII, and FV instead of fibrinogen, and thrombin time (TT) using only a small amount of thrombin can evaluate the intrinsic pathway and FVIII activity [16]. Recently, it was reported that CWA-TT can measure FVIII activity independent of the presence of emicizumab [17]. ...
... This study was conducted in accordance with the principles of the Declaration of Helsinki. The CWA-TT ( Figure 1a) using 0.5 IU thrombin (Thrombin 500 units; Mochida Pharmaceutical Co., Ltd., Tokyo, Japan) was measured using an ACL-TOP ® system (Instrumentation Laboratory, Bedford, MA, USA) [16,17]. This system shows three types of curves [16,17]. ...
... The CWA-TT ( Figure 1a) using 0.5 IU thrombin (Thrombin 500 units; Mochida Pharmaceutical Co., Ltd., Tokyo, Japan) was measured using an ACL-TOP ® system (Instrumentation Laboratory, Bedford, MA, USA) [16,17]. This system shows three types of curves [16,17]. One illustrates the changes in the absorbance observed while measuring CWA-TT, corresponding to the fibrin formation curve (FFC). ...
Background:
Regular prophylactic therapy has become an increasingly common treatment for severe hemophilia. Therefore, hypercoagulability-a potential risk factor of thrombosis-is a cause for concern in hemophilic patients treated with a high dose of FVIII concentrate. In clot waveform analysis (CWA)-thrombin time (TT), a small amount of thrombin activates clotting factor VIII (FVIII) instead of fibrinogen, resulting in FVIII measurements using CWA-TT with a small amount of thrombin.
Methods:
The coagulation ability of patients treated with FVIII concentrate or emicizumab was evaluated using activated partial thromboplastin time (APTT), TT and a small amount of tissue factor-induced FIX activation assay (sTF/FIXa) using CWA.
Results:
The FVIII activity based on CWA-TT was significantly greater than that based on the CWA-APTT or chromogenic assay. FVIII or FVIII-like activities based on the three assays in plasma without emicizumab were closely correlated; those in plasma with emicizumab based on CWA-TT and chromogenic assays were also closely correlated. CWA-APTT and CWA-TT showed different patterns in patients treated with FVIII concentrates compared to those treated with emicizumab. In particular, CWA-TT in patients treated with FVIII concentrate showed markedly higher peaks in platelet-rich plasma than in platelet-poor plasma. CWA-APTT showed lower coagulability in hemophilic patients treated with FVIII concentrate than in healthy volunteers, whereas CWA-sTF/FIXa did not. In contrast, CWA-TT showed hypercoagulability in hemophilic patients treated with FVIII concentrate.
Conclusions:
CWA-TT can be used to evaluate the thrombin bursts that cause hypercoagulability in patients treated with emicizumab. Although routine APTT evaluations demonstrated low coagulation ability in patients treated with FVIII concentrate, CWA-TT showed hypercoagulability in these patients, suggesting that the evaluation of coagulation ability may be useful when using multiple assays.
... However, few reports have described this relationship between the FVIII activity assessed using the peak time and height of CWA-APTT, including a small amount of tissue factor-induced activated FIX (sTF/FIXa) assay [14]. A CWA-small amount of thrombin time (CWA-TT) also reflects thrombin burst and FVIII activity [15] and can be used to measure the FVIII activity independent of the presence of emicizumab [16]. ...
... This study was carried out in accordance with the principles of the Declaration of Helsinki. The CWA-TT was measured using 0.5 IU thrombin (Thrombin 500 units; Mochida Pharmaceutical Co., Ltd., Tokyo, Japan) with an ACL-TOP ® system (Instrumentation Laboratory, Bedford, MA, USA) [15,16]. Three types of curves are shown on this system monitor [15,16]. ...
... The CWA-TT was measured using 0.5 IU thrombin (Thrombin 500 units; Mochida Pharmaceutical Co., Ltd., Tokyo, Japan) with an ACL-TOP ® system (Instrumentation Laboratory, Bedford, MA, USA) [15,16]. Three types of curves are shown on this system monitor [15,16]. One shows the changes in the absorbance observed while measuring the TT, corresponding to the fibrin formation curve (FFC). ...
Background: Although the use of regular replacement therapy including emicizumab for severe hemophilia has been spread, the assessment of the hemostatic ability using routine activated partial thromboplastin time (APTT) is still difficult in patients being treated with emicizumab. Methods: The hemostatic ability in patients treated with FVIII concentrate or emicizumab was evaluated by APTT, thrombin time (TT) and a small amount of tissue factor induced FIX activation assay (sTF/FIXa) using a clot waveform analysis (CWA). Results: FVIII activities based on a CWA-TT were significantly higher than those based on a CWA-APTT or chromogenic assay. FVIII activities based on the three assays in plasma without emicizumab were closely correlated, and those in plasma with emicizumab based on a CWA-TT and chromogenic assays were also closely correlated. The CWA-APTT and CWA-TT showed different patterns in patients treated with FVIII concentrates from those treated with emicizumab. In particular, the CWA-TT in patients treated with FVIII concentrate showed that the peak heights were significantly higher in platelet-rich plasma than in platelet-poor plasma. In plasma with approximately 16% of FVIII activity based on APTT assay from patients treated with FVIII concentrate, the peak height on the CWA-sTF/FIXa showed a higher hemostatic abilitythan normal plasma. Conclusions: The CWA-TT can measure the FVIII activity in patients treated with emicizumab. Although routine APTT evaluations demonstrate a low hemostatic ability in patients treated with FVIII concentrate, the CWA-TT and CWA-sTF/FIXa show hypercoagulability in those patients.
... Clot waveform analysis (CWA) [1][2][3][4][5] is based on the activated partial thromboplastin time (APTT) [1][2][3], prothrombin time (PT) [4], or thrombin time (TT) [5] (CWA-APTT, CWA-PT, and CWA-TT, respectively) ( Table 1). Although conventional clotting assays such as the APTT, PT, and TT are inexpensive, easy, and automated, enabling the measurement of multiple samples, they cannot visualize the clotting process. ...
... Clot waveform analysis (CWA) [1][2][3][4][5] is based on the activated partial thromboplastin time (APTT) [1][2][3], prothrombin time (PT) [4], or thrombin time (TT) [5] (CWA-APTT, CWA-PT, and CWA-TT, respectively) ( Table 1). Although conventional clotting assays such as the APTT, PT, and TT are inexpensive, easy, and automated, enabling the measurement of multiple samples, they cannot visualize the clotting process. ...
... Modified CWA is based on CWA-dilute PT [32,35], dilute TT [5], and clot-fibrinolysis waveform analysis (CFWA) [36,37]. CWAdilute PT shows both the extrinsic and intrinsic pathways. ...
Clot waveform analysis (CWA) observes changes in transparency in a plasma sample based on clotting tests such as activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). Evidence indicates that not only an abnormal waveform but also peak times and heights in derivative curves of CWA are useful for the evaluation of hemostatic abnormalities. Modified CWA, including the PT with APTT reagent, dilute PT (small amount of tissue factor [TF]-induced clotting factor IX [FIX] activation; sTF/FIXa), and dilute TT, has been proposed to evaluate physiological or pathological hemostasis. We review routine and modified CWA and their clinical applications. In CWA-sTF/FIXa, elevated peak heights indicate hypercoagulability in patients with cancer or thrombosis, whereas prolonged peak times indicate hypocoagulability in several conditions, including clotting factor deficiency and thrombocytopenia. CWA-dilute TT reflects the thrombin burst, whereas clot-fibrinolysis waveform analysis reflects both hemostasis and fibrinolysis. The relevance and usefulness of CWA-APTT and modified CWA should be further investigated in various diseases.
... Regarding COVID -19 (Figure 3), leukocyte counts are generally decreased early in COVID-19 [8], suggesting that activated platelets and injured vascular endothelial cells may play an important role in the onset of thrombosis through CD-147 [53]. However, CWA-APTT and CWA-sTF/FIXa showed hypercoagulability in patients with COVID-19 [41] suggesting that thrombin burst (Figure 2) [83] which is enhanced by activated platelets, causes hypercoagulability in this state. ...
Coronavirus disease 2019 (COVID-19) has spread, with thrombotic complications being increasingly frequently reported. Although thrombosis is frequently complicated in septic patients, there are some differences in the thrombosis noted with COVID-19 and that noted with bacterial infections. The incidence (6–26%) of thrombosis varied among reports in patients with COVID-19; the incidences of venous thromboembolism and acute arterial thrombosis were 4.8–21.0% and 0.7–3.7%, respectively. Although disseminated intravascular coagulation (DIC) is frequently associated with bacterial infections, a few cases of DIC have been reported in association with COVID-19. Fibrin-related markers, such as D-dimer levels, are extremely high in bacterial infections, whereas soluble C-type lectin-like receptor 2 (sCLEC-2) levels are high in COVID-19, suggesting that hypercoagulable and hyperfibrinolytic states are predominant in bacterial infections, whereas hypercoagulable and hypofibrinolytic states with platelet activation are predominant in COVID-19. Marked platelet activation, hypercoagulability and hypofibrinolytic states may cause thrombosis in patients with COVID-19.
... An optical automatic coagulation analyzer has been able to demonstrate the clot reaction curve of APTT [14,15], diluted prothrombin time (PT) [16] and thrombin time (TT) [17]. Such an analysis of the coagulation curve is called a clotting waveform analysis (CWA) [18]. ...
... In addition, sTF/FIXa may reflect thrombin burst, which mainly depends on the activation of FXI, FVIII and FV [20,21]. CWA-TT has also been reported to be useful for evaluating thrombin burst [17]. Although measurement of FVIII activity in patients treated with emicizumab using anti-idiotype, monoclonal antibodies is possible [22], this method is not easy to perform. ...
... The CWA-TT was measured using 0.5 IU thrombin (500 units of thrombin, Mochida Pharmaceutical Co., Ltd., Tokyo, Japan) with an ACL-TOP ® system (Instrumentation Laboratory, Bedford, MA, USA) [17]. Three types of curves are shown on this system monitor [21]. ...
Objective:
Although emicizumab is a bispecific, monoclonal antibody that has led to a significant improvement of treatment for hemophilia A patients with inhibitors, the routine monitoring of patients treated with emicizumab is difficult. Thrombin time (TT) reflects thrombin burst, which mainly depends on activation of factor V (FV) and FVIII.
Methods:
We, therefore, developed a method for evaluating clotting activity independent of the presence of emicizumab. Normal plasma (NP) or FVIII-deficient plasma (FVIIIDP) with and without emicizumab was measured using clot waveform analysis (CWA)-activated partial thromboplastin time (APTT) and TT.
Results:
Emicizumab caused clot formation in FVIIIDP using the CWA-APTT; however, the coagulation peaks of plasma with and without emicizumab measured by the CWA-TT did not differ to a statistically significant extent. Regarding the mixing tests with NP and FVIIIDP, CWA-APTT showed large differences between each mixing test in plasma with and without emicizumab, whereas the CWA-TT showed similar patterns in mixing plasma with and without emicizumab. Regarding the standard curve of FVIII activity, the CWA-APTT showed an FVIII-concentration-dependent increase; however, the values with each concentration of FVIII differed between samples with and without emicizumab, whereas CWA-TT showed FVIII-concentration-dependent fluctuations independent of the presence of emicizumab, and the values with each concentration of FVIII were similar in samples with and without emicizumab.
Conclusions:
As CWA-TT using a small amount of thrombin (0.5 IU/mL) can reflect thrombin burst and be useful for evaluating FVIII activity, independent of the presence of emicizumab, it is useful for monitoring clotting activity in patients with an anti-FVIII inhibitor treated with emicizumab.
