Στο Διεθνές συνέδριο της Αγγεοχειρουργικής και Αγγειολογίας που γίνεται στην Αθήνα τον Ιούνιο έστειλα τις ακόλουθες τρεις εργασίες
RECURRENT FETAL LOSSES: ETIOLOGICAL AND THERAPEUTIC APPROACH
Michael P Makris, Athanasios Kanidis Pantelis E Makris
Aristotle University of Thessaloniki- Greece.
Many researchers, beside the usual causes, implicate disturbances in haemostasis (haemorrhagic or hyper coagulant) for recurrent fetal losses. It appears that during the 1st trimester of pregnancy fetal losses occur that are connected mainly with haemorrhagic diathesis (such as minor Willebrand disease or other minor coagulant factors deficiencies), while fetal losses between the 2nd and 3d trimester are connected with thrombophilic diathesis (such as hereditary or acquired thrombophilia e.g. antiphospholipid syndrome).
In the last three years we examined 91 women with recurrent fetal losses in order to determine the causes of this losses. We followed up their courses applying if possible anticoagulant treatment in the cases that were connected with hereditary or acquired thrombophilic diathesis and on the cases that haemorrhagic diathesis was responsible we treated with DDAVP to control this diathesis.
In order to make an estimate on the success of the treatment we applied on these cases, we made a record.
Material: we studied 91 women with mean age 33,9 ±4,8 years that had about 2,9±1,3 fetal loses each.
Method: We studied all the possible factors of haemostasis that can be implicated for disturbances with the automatic instruments of DADE-Behring and IL (Advance) and also full genetic control with the strip assay of Vienna Lab with extracted DNA by Qiagen kit.
Results: Amongst our results we found 11 women heterozygote for FV-Leiden, 9 women were carriers of the ΗΡΑ-1 (Gp-IIIa) mutation and also 3 more women had the FV -HR2 with the ΗΡΑ-1 (Gp-IIIa) mutation, 1 woman with FII 20210 G>A mutation, 2 women with the 455 G>A mutation of fibrinogen. Also 2 more women with protein S deficiency, 1 woman with FXII deficiency, 1 woman showed plasminogen deficiency, 3 women with antiphospholipid syndrome and 8 women had haemorrhagic diathesis (mainly mild von Willebrand disease). Overall in 41 women we found risk factors connected with recurrent fetal losses. From those women 29 were able to have one effective pregnancy with the help of antithrombotic treatment (μΜΒΗ, or antipllatelets mainly aspirin) or with DDAVP in order to increase the levels of factor Willebrand. In the other women an effective therapeutic approach was not possible and they seized their attempts.
Comment: The determination of the causes of recurrent fetal losses by a hematologist, who can make a full functional and genetic control of the haemostasis, can help in an effective pregnancy.
ORAL ANTICOAGULATION : DETECTION OF VKORC1 GENE MUTATIONS IN GREEKS PATIENTS
Makris MP, Amiridou B, Koliakos G, Makris PE
Aristotle University of Thessaloniki-Greece
The oral anticoagulant treatment with warfarin is responsible for the 10% of all submissions in hospitals due to the side affects that warfarin causes. Those side affects are shown as minor or major haemorrhagic complications or as recurrent thromboembolic episodes. For these side affects factors like age, height, nutrition habits, interaction with other drugs and genetic mutations have been found guilty for increasing the sensitivity in warfarin and also the resistance against it. For the resistance against warfarin the mutation of the epoxidasse of vitamin K (VKORC1) and its polymorphisms were found guilty, whose presence blocks the action of warfarin as an anticoagulant.
Aim: The study was made in order to detect the mutation of VKORC1 in patients under antivitamin K treatment.
Material: we studied 145 subjects (105 were the control group without receiving any anticoagulant and 40 patients receiving antivitamin K)
Mtehod: DNA extractions from whole blood with Qiagen kit, Real-Time PCR with SYBR Green in order to detect the presence of any mutation in the gene of VKORC1. We also checked clinical information (increased sensitivity in small doses of Sintrom) with the strip assay of ViennaLab that detectes the polymorphism VKORC1 1639 G>A.
Results: From the RT PCR we did not have any important findings due to the fact that the mutations of VKORC1 are rare.
The use of the specific for the 1639 G>A polymorphism strip assay of ViennaLab gave one homozygote AA patient (explaining the increased sensitivity in Sintrom 1mg5 INR)
This finding resulted in the isolation of the melting curve of this patient which led us to discover for the first time the very low fluorescence in comparison with a normal subject.
Comment: if we are able to detect the mutation responsible for the increased resistance in antivitamin K and the polymorphism of VKORC1 who relates with the increased sensitivity can help us determine problems in the treatment with antivitamin K and the follow up of the treatment
και τέλος την 3η εργασία:
ORAL ANTICOAGULANT: STUDY OF THE C1173T POLYMORPHISM OF VKORC1 IN THROMBOPHILIC PATIENTS.
Karanastasi S, Makris MP, Kalogeridis A, Koliakou K, Koliakos G, Makris PE
Aristotle University of Thessaloniki and Biohellenika co. Greece
Backround : The oral treatment with antivitamin K is being applied in clinical practice almost exclusively for the prevention and treatment of thromboembolic disease. This treatment often comes with haemorrhagic or thrombotic complications that are connected with the right adjustment of the dosage and the follow up of the treatment due to number of factors that are involved such as age, body weight, nutrition habits and genetic factors. It has been observed that the haemorrhagic problems appear much more often. When the research for the genetic causes that affect the adjustment of the antivitamin K treatment took place the polymorphism VKORC1 1173 C>T and mainly the homozygote form (1173TT) was found to be connected with frequent haemorrhagic complications due to the fact that the homozygote have increased sensitivity in low doses of antivitamin K. On the contrary the patients with 1173CC form require at least the double dosage of antivitamin K to reach the wanted results (INR=2.5)
Aim: Because of these observations we decide to check this polymorphism in thrombophilic patients in oral anticoagulant treatment.
Material: We studied 194 subjects from which 49 were normal (28 male, 21 female) and 135 thrombophilic patients (71 male, 64 female). All our thrombophilic patients had a 2 year follow up.
Methods: DNA extraction with the Qiagen kit, we applied conventional PCR technology with TIB MOLBIOL scheduled primers and the products were digested with Hinf 1 restriction enzyme from Roche Molecular Biochemicals. The digestion products were studied with electrophoresis in agarose 2% gell.
Results: From the 49 normal subjects we found out that 27 of them were heterozygote, 12 homozygote CC and 10 homozygote TT (fig). From the 135 thrombophilic patients we found 28 homozygote TT (who were treated with 2.5±0.69 mg of antivitamin K daily), 71 heterozygote CT (who were treated with 3.5±0.9 mg daily), and 36 homozygote CC (who were treated with 4.25±0.9 mg daily). In conclusion we found out that indeed the homozyte TT needed a smaller dosage of antivitamin K in comparison with the heterozygote CT and the CC homozygote. We note that our patients did not show any haemorrhagic or thrombotic complications.