Circulating activated protein C levels are not increased in septic patients treated with recombinant human soluble thrombomodulin

Figures

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  Fig. 1. rTM promotes thrombin-mediated APC generation in vitro. Pooled normal plasma was incubated with thrombin (0–0.2 U/mL) and rTM (0–2000 ng/mL) at 37 °C for 10 min. APC levels were analyzed by the OLIGOBIND APC activity assay. The data represent mean ± SD (n = 3). Representative data of two independent experiments are shown

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  Fig. 2. Decreased concentrations of protein C and antithrombin are associated with decreased and increased APC generation, respectively. a Plasma samples with 0, 10, 30, 50, 70, and 100% protein C levels were prepared by the addition of vehicle, 0.36, 1.08, 1.81, 2.53, and 3.61 μg/mL of protein C, respectively, to protein C deficient plasma. b Plasma samples with 0, 10, 30, 50, 70, and 100% antithrombin levels were prepared by the addition of vehicle, 0.07, 0.25, 0.43, 0.61, and 0.88 U/mL of antithrombin, respectively, to antithrombin deficient plasma. These plasma samples were incubated with 0.2 U/mL of thrombin (T1063, Sigma-Aldrich, St. Louis, MO) and 1000 ng/mL of rTM at 37 °C for 10 min. APC levels were analyzed by the OLIGOBIND APC activity assay. The data represent mean ± SD (n = 3). Representative data of two independent experiments are shown. PNP: pooled normal plasma, PC: protein C, AT: antithrombin

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  Fig. 3. Plasma APC levels are not increased in septic patients during a 30–60-min period of rTM treatment, while TAT levels are decreased during the treatment period. a Blood samples were collected from eight patients with sepsis-associated DIC before and after administration of rTM (130 U/kg: red symbols; 380 U/kg: blue symbols) on day 1 and day 2. Plasma sTM levels were determined by ELISA. b Plasma TAT levels were analyzed by STACIA CLEIA TAT. c Plasma APC levels were analyzed by the OLIGOBIND APC activity assay. d Plasma protein C levels were measured by a synthetic chromogenic substrate method using HemosIL Protein C. A paired t-test was used for comparisons before and after rTM treatment. *P < 0.05, **P < 0.01