Novel starches: single-dose pharmacokinetics and effects on blood coagulation.

BACKGROUND: Carboxymethyl starch (CMS) and carboxymethylated hydroxyethyl starch (CM-HES) might offer advantages over hydroxyethyl starch (HES) with regard to their volume expansion effect and their pharmacokinetic characteristics. The goal of the current study was to determine the pharmacokinetics of CMS and CM-HES and to investigate their influence on blood coagulation in comparison with the standard low-molecular, low-substituted HES (130/0.42) used in Europe. METHODS: The study was conducted as a randomized, blinded, parallel three-group study in 30 pigs. Twenty ml/kg of 6% HES (control), 6% CMS, or 6% CM-HES was infused as a single dose, and serial blood sampling was performed over 20 h to measure plasma concentration and molecular weight and to assess blood coagulation. Concentration-effect relations were assessed by pharmacokinetic-pharmacodynamic analysis. RESULTS: CMS and CM-HES showed significantly higher plasma concentrations and molecular weights over 20 h (P for both<0.001) with smaller volumes of distribution and longer elimination rates during the terminal phase (P for both<0.01) when compared with HES. CMS and CM-HES impaired whole blood coagulation more than HES as assessed by Thrombelastograph analysis (Haemoscope Corporation, Niles, IL). However, similar effects of all three starch preparations on blood coagulation were found when related to the plasma concentrations in mass units. CONCLUSIONS: Carboxymethylation of starch results in an increased intravascular persistence and a slower fragmentation compared with HES. The greater impairment of blood coagulation by CMS and CM-HES seems to be caused by the higher plasma concentrations.

Novel, semi-automated, 60-min-assay to determine von Willebrand factor cleaving activity of ADAMTS-13.

INTRODUCTION: The clinical entity of idiopathic thrombotic thrombocytopenic purpura (TTP) has been closely associated with a severe deficiency of the von Willebrand factor (vWF)-cleaving protease (ADAMTS-13). Levels below 5% are highly specific for TTP basically excluding hemolytic uremic syndrome (HUS). Deficiency of ADAMTS-13 can either be inherited or caused by inhibitory IgG autoantibodies to ADAMTS-13. Diagnosis of TTP is often difficult since it often manifests oligosymptomatic exhibiting only laboratory signs of thrombocytopenia and hemolysis or involving atypical organs. In such cases rapid knowledge of the ADAMTS-13 activity is very useful in helping to establish a correct diagnosis rapidly and to initiate the appropriate therapy. METHODS AND RESULTS: We have therefore designed a rapid, sensitive and simple assay for the determination of ADAMTS-13 activity and ADAMTS-13 inhibitors. The test is based on the measurement of the ristocetin-cofactor activity of purified, urea-denaturated vWF concentrate after incubation with diluted patient plasma that has been preactivated with barium chloride. While this assay has already previously been established, we have further modified and simplified the test thereby improving markedly the sensitivity, accuracy and turn-around time. A major improvement proved to be the change from buffer to heat-treated and vWF-replenished plasma as diluent for making the serial dilutions of normal pool plasma (NPP) for the calibration curve. Set up as a semi-automated assay the test is performed in only 50 to 60 min. Tested in 15 patients with ADAMTS-13 deficiency this new assay compared favorably and even more accurately with the classic electrophoretic ADAMTS-13 assay, especially in the low range below 10%. Mixing studies with patient plasma and NPP (ratio 1:1) revealed that a relatively short period of only 30 min at room temperature is sufficient for most samples to detect an inhibitor. With a single mixing experiment it is possible to discriminate inhibitor levels in the range of 0.5-4 Bethesda units. CONCLUSION: Using this novel assay, determination of ADAMTS-13 activity and ADAMTS-13 inhibitors can be performed easily, rapidly (less than 2 h), and accurately, providing rapid information about the severity of ADAMTS-13 deficiency and the presence of an inhibitor. In addition, the test is suitable for full automation enabling to screen large populations and, thus, help to get further insights into the clinical significance of ADAMTS-13 deficiency in other diseases.

The effect of clonidine on perioperative blood coagulation.

STUDY OBJECTIVE: To evaluate the influence of perioperative stress protection by clonidine on blood coagulation. DESIGN: Prospective, randomized, double-blinded, placebo-controlled clinical trial. SETTING: University hospital. PATIENTS: 50 patients scheduled for elective gynecoabdominal surgery. INTERVENTIONS AND MEASUREMENTS: Patients were randomly assigned to control (placebo) or clonidine group (single intravenous clonidine dose; 4 microg/kg(-1) or 3 microg/kg(-1) for age >65 years). Three measurement time points were defined: before administration of placebo/clonidine and anesthesia induction, (t1; baseline measurement); after surgery, before emergence of anesthesia (t2); and at the first postoperative day, 24 hours after anesthesia induction (t3). Blood coagulation was analyzed at all time points measuring international normalized ratio, platelets, thrombin-antithrombin complex, von Willebrand factor, soluble thrombomodulin, d-dimers, plasminogen activator inhibitor 1, and Thrombelastograph analysis. MAIN RESULTS: In the postoperative period (t2, t3), hypercoagulability was present in all patients compared with baseline measurements (t1) but without differences between the control and clonidine group. Regarding hematologic, laboratory blood coagulation, and Thrombelastograph parameters, there was no statistically and clinically relevant difference throughout the study period between the 2 groups. No hemodynamic adverse events of clonidine were observed in the perioperative period. Until day of discharge, no thrombotic or thromboembolic events were reported in both groups. CONCLUSIONS: Preoperative administration of a single dose of clonidine has no effect on perioperative blood coagulation.

Decreased anticoagulant response to tissue factor pathway inhibitor in patients with venous thromboembolism and otherwise no evidence of hereditary or acquired thrombophilia.

No relevant deficiency of TFPI or genetic polymorphisms could thus far consistently be associated with venous thromboembolism. We hypothesized that the substrates of the TFPI protein, including FVII or FX (rather than the protein itself) could induce a hypercoagulable state. We created a novel TF-based clotting assay that evaluated the anticoagulant response to exogenously added recombinant TFPI. The response to TFPI was expressed as the ratio of the clotting time with and with-out TFPI. By using 118 healthy controls, we established a reference range between 1.31 and 1.93 (mean value +/-2 standard deviations (SD), 1.62 +/-0.31). We then evaluated samples from 120 patients with a history of venous thromboembolism but no evidence of hereditary and acquired thrombophilia. The range Coagulation Laboratory, Division of Haematology, University Hospital of Zurich, Zurich, Switzerland of the patients' ratios was significantly (P < 0.001) lower, falling between 1.2 and 1.78 (mean value +/-2 SD, 1.49 +/-0.29). Of the 120 patients, 39 (32.5%) had a TFPI sensitivity ratio below the 10th percentile of the controls, compared with 11 (9.3%) of the healthy controls. The crude odds ratio for venous thrombosis for subjects with a TFPI sensitivity ratio below the 10th percentile was 13 (95% CI; range, 3.1 to 54.9) compared with those with a ratio above 1.8 (90th percentile). Patients with idiopathic thromboembolism did not have a decreased TFPI sensitivity ratio more often than patients with thrombosis with a circumstantial risk factor. Based on these results, a reduced response to TFPI may lead to an increased risk of venous thrombosis.

Prevalence of hereditary thrombophilia in patients with thrombosis in different venous systems.

The prevalence of hereditary thrombophilia is well known in patients with lower-extremity thrombosis but only poorly studied in patients with thrombosis at unusual sites. Consequently, it is still unclear whether such patients should generally be screened for hereditary thrombophilia. We retrospectively analyzed 260 patients with thrombosis at unusual sites including thrombosis in portal, cerebral, retinal, and upper-extremity veins with respect to the prevalence of FV Leiden, prothrombin G20210A, protein C, protein S, and antithrombin deficiency. In addition, all thrombotic episodes were analyzed for circumstantial risk factors. Used as controls, healthy volunteers (120) and patients with lower-extremity thrombosis (292) showed overall prevalence of hereditary thrombophilia of 9.1% and 39.0%, respectively. The corresponding numbers were 33.3%, 34.3%, and 39.0% in patients with portal vein, upper-extremity, and lower-extremity thrombosis, respectively. In patients with cerebral vein thrombosis, however, the prevalence was significantly lower (23.5%). Patients with retinal vein occlusion did not show an increased frequency of thrombophilia at all (5.9%). In all five groups FV Leiden was by far the most frequent defect (4.4-27.1%), while prothrombin G20210A occurred rarer (2.5-7.6%). Protein C, protein S, and antithrombin deficiency were much less prevalent (0-3.1%) except for patients with portal vein thrombosis (4.8-7.1%). Compared to healthy individuals, the relative risk of thrombosis was 4.3 (2.2-8.1), 3.8 (1.8-7.7), 2.5 (1.0-6.1), 3.7 (1.5-8.6), and 0.6 (0.2-2.1) for patients with lower-extremity, upper-extremity, cerebral vein, portal vein, and retinal vein thrombosis, respectively. Circumstantial risk factors were more frequent in patients without than with hereditary thrombophilia and were found most often in patients with upper-extremity thrombosis. In each group the most frequent circumstantial risk factor was different. However, oral contraceptives and cancer were found in all five groups. In conclusion, independent upon the presence of circumstantial risk factors, screening for hereditary thrombophilia is warranted in all patients with thrombosis at unusual sites except in those with retinal vein occlusion.