Clinical Application of a Chromogenic Substrate Method for Determination of Factor VIII Activity

Rosén, Steffen; Andersson, Mikael; Blombäck, Margareta; Hägglund, U; Larrieu, M.J.; Wolf, Martine; Boyer, Catherine; Rothschild, Chantal; Im, Nilsson; Sjörin, Elsy

doi:10.1055/s-0038-1660140

Cited by 59 publications

(37 citation statements)

References 12 publications

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“…Cleav age of FPA from fibrinogen has been shown to occur with thrombin concentrations about 100 times less than do release BTG from platelets [12], Whether thrombin activ ity may injure platelets prior to detectable release of BTG is not known. With regard to FVIIFC, the chromogenic peptide substrate assay employed in this study is insensi tive to activation of FVIIFC by thrombin [19], and such activation would, therefore, have escaped our detection. One could, however, have expected to detect a faster decay of FVIIFC following storage of blood with high FPA levels, but this was not the case.…”

Section: Resultsmentioning

confidence: 96%

Thrombin Generation During Collection of Blood from Donors Taking Oral Contraceptives

Skjønsberg¹,

Kierulf²,

Engebretsen³

et al. 1987

Vox Sanguinis

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Thrombin generation, as evidenced by plasma fibrinopeptide A (FPA) concentrations, was studied during blood collection from donors taking oral contraceptives (OC). 450 ml blood were drawn into Fenwal PVC bags from 26 OC users and 28 nonusers. Blood samples for determination of FPA, beta-thromboglobulin (BTG), thrombotest (TT), prekallikrein (PKK), antithrombin-III (AT-III) and factor VIII procoagulant activity (FVIII:C) were drawn from the bags immediately after ending blood donation and following storage for 24 h at 4°C. The FPA concentrations following donation were significantly higher in the OC than in the control group (p<0.05). The levels of PKK were also higher in blood obtained from OC users (p<0.001), as was the FVIII:C level, the latter difference, however, was not significant (p=0.06). No cold-promoted activation of factor VII, as evidenced from TT, was detected following storage at 4°C, neither was any change observed in the FPA, PKK and AT-III levels. The BTG concentrations increased significantly during storage, most pronounced in the control group (p<0.05). The decay of FVIIIiC was similar in the two groups, averaging 24.7%. No correlation was observed between the FPA levels and the other parameters determined. We conclude that thrombin generation is more pronounced during routine blood collection from donors taking OC.

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Section: Resultsmentioning

confidence: 96%

Thrombin Generation During Collection of Blood from Donors Taking Oral Contraceptives

Skjønsberg¹,

Kierulf²,

Engebretsen³

et al. 1987

Vox Sanguinis

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“…However, in a recent paper on the effects of high FVIII levels on the risk of VTE recurrence, they measured FVIII using both methods and could not confirm that the chromogenic method gives more reproducible results than the clotting method, as has been reported previously. [23][24][25][26][27] Third, because FVIII levels were only measured once, it is possible that misclassification occurred due to a transient rise in FVIII as an acute reaction. In order to avoid this error, the blood samples were taken from resting subjects by an experienced technician.…”

Section: Resultsmentioning

confidence: 99%

High Plasma Level of Factor VIII - An Important Risk Factor for Venous Thromboembolism -

Ota

Yamada

Ogihara

et al. 2011

Circ J

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Background: Elevated levels of Factor VIII (FVIII) have been suggested as important for the pathogenesis of venous thromboembolism (VTE). The objective of this study was to explore the association between elevated FVIII level and VTE in Japan. Methods and Results:68 patients with objectively documented VTE and 40 controls were included. In the patients, the FVIII level was measured for a mean follow-up period of 52.6 months (3-348 months) after the onset of VTE, in order to avoid an acute-phase response. The VTE patients had a higher mean FVIII level than the controls (154.5±55.8 IU/dl vs. 114.3±16.0 IU/dl, P<0.0001). According to multivariate analysis, FVIII levels above the 50 th percentile (124 IU/dl) conferred an odds ratio of 4.9 (95% confidence intervals (CI) 1.9-12.4; P<0.001) and those above the 75 th percentile (150 IU/dl) conferred an odds ratio of 31.9 (95%CI 4.0-252.3; P<0.001). Conclusions:Elevated plasma FVIII level is a risk factors for VTE in Japanese people, and FVIII levels above the 75 th percentile are associated with a significant odds ratio for the occurrence of VTE. (Circ J 2011; 75: 1472 - 1475

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“…However, at least for fVIII, this increase was not observed in a preactivation-insensitive two-stage activity assay (Table 2). 21 Similarly, fVIII antigen levels by ELISA were comparable in AT ϩ/ϩ and AT m/m mice (500Ϯ100 ng/mL for AT m/m versus 410Ϯ230 for AT ϩ/ϩ mice; meanϮSD, nϭ4 to 8, PϭNS). These results suggested a low level continuous activation of the coagulation system in AT m/m mice, consistent with the observed thrombotic phenotype.…”

Section: Hemostasis Parameters In Adult Wild-type and Mutant Micementioning

confidence: 88%

“…Plasma fibrinogen was determined by a coagulation rate assay. 20 Factor VIII plasma levels were additionally determined using the Coatest Factor VIII kit (Chromogenix, Sweden) 21 and by ELISA. 22 Plasma AT antigen levels were determined by rocket immunoelectrophoresis 23 using a polyclonal rabbit anti-human AT antibody (Dako) that cross-reacts with murine AT.…”

Section: Coagulation and Hematological Parametersmentioning

confidence: 99%

Life-Threatening Thrombosis in Mice With Targeted Arg48-to-Cys Mutation of the Heparin-Binding Domain of Antithrombin

Dewerchin

Hérault

Wallays

et al. 2003

Circulation Research

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Abstract-Antithrombin (AT) inhibits thrombin and some other coagulation factors in a reaction that is dramatically accelerated by binding of a pentasaccharide sequence present in heparin/heparan-sulfate to a heparin-binding site on AT.Based on the involvement of R47 in the heparin/AT interaction and the frequent occurrence of R47 mutations in AT deficiency patients, targeted knock-in of the corresponding R48C substitution in AT in mice was performed to generate a murine model of spontaneous thrombosis. The mutation efficiently abolished the effect of heparin-like molecules on coagulation inhibition in vitro and in vivo. Mice homozygous for the mutation (AT m/m mice) developed spontaneous, life-threatening thrombosis, occurring as early as the day of birth. Only 60% of the AT m/m offspring reached weaning age, with further loss at different ages. Thrombotic events in adult homozygotes were most prominent in the heart, liver, and in ocular, placental, and penile vessels. In the neonate, spontaneous death invariably was associated with major thrombosis in the heart. This severe thrombotic phenotype underlines a critical function of the heparin-binding site of antithrombin and its interaction with heparin/heparan-sulfate moieties in health, reproduction, and survival, and represents an in vivo model for comparative analysis of heparin-derived and other antithrombotic molecules.

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Clinical Application of a Chromogenic Substrate Method for Determination of Factor VIII Activity

Cited by 59 publications

References 12 publications

Thrombin Generation During Collection of Blood from Donors Taking Oral Contraceptives

Thrombin Generation During Collection of Blood from Donors Taking Oral Contraceptives

High Plasma Level of Factor VIII - An Important Risk Factor for Venous Thromboembolism -

Life-Threatening Thrombosis in Mice With Targeted Arg48-to-Cys Mutation of the Heparin-Binding Domain of Antithrombin

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