Page 47 - Pro-active Management of Women’s Health after Cardiometabolic Complicated Pregnancies

2.2 Associations between phenotypes of preeclampsia and thrombophilia| 45
diluted Prothrombin time (Recombiplastin, Instrumentation Laboratories, Italy, normal ratio < 1.20) and the activated partial thromboplastin time assay (Platelin, Organon Teknika, USA, normal value < 40 seconds). Effect of factor deficiency was eliminated by mixing 1:1 patient and normal pool plasma. Protein S (normal range 0.70-1.40 U/ml) was determined as protein S activity by a clotting assay (Diagnostica Stago, France) and as free protein S by an ELISA (Biopool, USA). Protein S deficiency is defined as an outcome of <0.70 (IU/ ml) for two assays at least 6 weeks apart without use of oral contraceptives. Protein C (normal range 0.70-1.40 U/ml) deficiency was determined by measuring protein C activity (IL Testkit ProclotTM Proclot, Instrumentation Laboratory, Italy) and protein C antigen by a house-made ELISA with the use of rabbit antihuman anti-protein C (Dako Cytomation, Denmark). Protein C deficiency is defined as an outcome of <0.70 (IU/ ml) for two assays at least 6 weeks apart.
Serum homocysteine concentrations were measured both after (an overnight) fasting and 6 hours after an oral loading dose of L- methionine (0.1 g/kg body weight). During the test the patients were given a standardized methionine-poor breakfast and lunch. EDTA-blood samples were kept on ice and plasma was separated within one hour for determination of homocysteine. Plasma total homocysteine was determined using HPLC with fluorescence detection. The patients were considered to have hyperhomocysteineamia if the fasting homocysteine concentration exceeded 15 μmol/L and/or the post-load value exceeded 45 μmol/L.
Factor V Leiden and Prothrombin gene mutation were determined by polymerase chain reaction.
Patients with an abnormal test result, except for factor V Leiden, Prothrombin gene mutation, and serum homocysteine concentrations were retested at least six weeks after the initial test. Only if the same abnormality was