strategies used to investigate suspected VTE in postpartum individuals.Future prospective studies should aim to improve the postpartum diagnosis of VTE.

AIMS: We aimed to characterize relationship between the expression profi les of platelet miR-96, miR-126 and miR-223 and platelet function examination in patients with sticky platelet syndrome (SPS) and in healthy controls.BACKGROUND: MicroRNAs (miRNA, miR) are a group of small and non-coding RNAs involved in many mechanisms as regulators of post-transcriptional protein expression in platelets.SPS is defi ned as platelet hyperaggregability after administration of low doses of adenosine diphosphate and/or epinephrine.Clear genetic abnormality of this syndrome is not known yet.METHODS: We examined 45 patients with SPS and 30 healthy volunteers.For functional platelet examination we used light transmission aggregometry, and qRT-PCR was used to determine the expression of the miRNAs.RESULTS: We observed no relationship of the platelet miRNA expression with functional platelet examination in the entire cohort of patients with SPS.However, in a group of patients with SPS and pregnancy complications, we found that the expression of platelet miR-96 (p = 0.009) was up-regulated.CONCLUSION: In spite of the multiple limitations of the study, it can be considered that the increased expression of platelet miR-96 found in a group of patients with SPS and pregnancy complications could be related to the hyperaggregability in these selected patients (Tab.2, Ref. 31).

Abstract Introduction: Sticky platelet syndrome (SPS) is referred to as a platelet hyperaggregability triggered by low concentrations of platelet agonists adenosine diphosphate (ADP) and/or epinephrine (EPI). Platelet aggregation with other inducers (collagen, arachidonic acid, ristocetin, and thrombin) remains within a normal range. MicroRNAs (miRNAs) are small, non-coding RNA molecules that play an important role in post-transcriptional regulation of protein expression. More recently, several studies show that the platelets are an abundant source of miRNAs and that the miRNA expression profiles within platelets correlate with the platelet reactivity. Aim: The principle objective of this article is to describe the method which we developed for the preparation of the pure platelet samples and report the results of this method. These final pure platelet samples are intended to be the first step for the platelet miRNA testing. Methods: The blood samples from 50 subjects were examined in the study. Then, the platelet rich plasma (PRP) samples obtained by centrifugation of the patient blood samples were used for our experiments. Subsequently, the erythrocytes and leucocytes remaining in PRP sample were magnetically labelled by CD45 Microbeads and CD235a Microbeads. After incubation the PRP sample passed through the magnetic separation system and the magnetically labelled cells (erythrocytes and leucocytes) were retained within the column of separator. The number of cells in the final PRP samples was measured by the blood cell analyser. Results and conclusion: We successfully developed and optimized the effective and reproducible method for magnetic separation of platelets, resulting in the leukocyte-depleted and erythrocyte-depleted platelet samples, which can be used for further genetic analyses.

International Journal of Gynecology & ObstetricsVolume 107, Issue S2 p. S458-S458 Poster presentations P164 Circulating VEGF serum profile during pregnancy in prediction of preeclampsia – is it helpful? K. Biskupska Bodova, K. Biskupska BodovaSearch for more papers by this authorK. Dokus, K. DokusSearch for more papers by this authorP. Zubor, P. ZuborSearch for more papers by this authorJ. Ivankova, J. IvankovaSearch for more papers by this authorJ. Stasko, J. StaskoSearch for more papers by this authorJ. Danko, J. DankoSearch for more papers by this author K. Biskupska Bodova, K. Biskupska BodovaSearch for more papers by this authorK. Dokus, K. DokusSearch for more papers by this authorP. Zubor, P. ZuborSearch for more papers by this authorJ. Ivankova, J. IvankovaSearch for more papers by this authorJ. Stasko, J. StaskoSearch for more papers by this authorJ. Danko, J. DankoSearch for more papers by this author First published: 20 November 2009 https://doi.org/10.1016/S0020-7292(09)61655-6AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume107, IssueS2Abstracts of XIX FIGO World Congress of Gynecology and ObstetricsOctober 2009Pages S458-S458 RelatedInformation

Patients with hemophilia demonstrate quite variable clinical phenotype even in cases with the same level of deficient factor or the same molecular abnormality. Different interacting factors including congenital and acquired alterations of coagulation inhibitors can modulate both clinical expression and severity of hemophilia. In this study, plasma levels of factor VIII (FVIII), factor IX (FIX) as well as protein C (PC), protein S (PS), and antithrombin (AT) plasma levels were measured in 80 patients with severe hemophilia A and B. Patients were divided into two groups according to the risk of bleeding: the first group (n = 32) with mild bleeding (< 2 bleeds/year), and the second group (n = 48) with severe bleeding (≥ 2 bleeds/year). Both hemophilia groups showed significantly decreased PC plasma levels compared to levels in healthy control subjects (the first group: p < 0.0001 and second group: p < 0.01). The difference in PC plasma levels between the first and second hemophilia group was significant (p < 0.05). Moreover, there was positive correlation between age and the functional PC in both hemophilia groups. Our results suggest that decreased PC plasma levels can testify to a slightly protective effect of the PC pathway on the severity and frequency of bleeding in patients with severe hemophilia A and B.

We read with a great interest the article from J. A. Heit et al. [1]. As was mentioned in the article, thrombomodulin has an important anticoagulant role in regulation of hemostasis and thus mutations causing its dysfunction or impaired synthesis could result in a prothrombotic state [2, 3]. One of the point mutations discussed in the article, the 127G to A substitution, was reported to be a risk factor for arterial thrombosis, namely for the thrombosis of coronary arteries. In the Study of Myocardial Infarctions Leiden (SMILE), the mutation was identified in 12 of 560 patients with myocardial infarction [4]. It was concluded that the presence of the mutation elevated the risk of myocardial infarction especially in men younger than 50 years and in individuals with additional risk factors. The relation between the mutation and venous thromboembolism was not so clearly defined, although several cases of patients with the 127G to A substitution and venous thrombosis were reported [5, 6]. Interestingly, the results of several relatively large studies suggested that the A allele is rather infrequent finding, even in a population of patients with myocardial infarction [1, 4, 7, 8]. The authors of SMILE examined altogether 1206 individuals (560 patients with myocardial infarction, 646 healthy controls) and identified 12 (2.1%) and 7 (1.1%) heterozygous carriers of the A allele [4]. J. A. Heit et al. found two heterozygotes among 208 patients with venous thromboembolism and two heterozygotes among 220 healthy controls [1]. Both studies failed to identify a homozygous carrier of the A allele. Warner et al. [8] examined the population of 465 patients with acute stroke and 353 healthy individuals and estimated the frequency of the A allele to 0.5% and 0.7%, respectively [8]. We tested the presence of 127G to A mutation in 244 patients (78 men with mean age 43.9 ± 14.4 years and age range 15–69 years and 166 women with mean age 40.7 ±12.2 years and age range 16–75 years) with a positive history of arterial and/or venous thromboembolic events and negative thrombophilic screening. A total of 184 patients (49 men and 135 women; 26 men and 78 women with the first event) suffered from venous thromboembolism, 52 patients (23 men and 29 women; 19 men and 15 women with the first event) suffered from arterial thromboembolism and eight patients (all women with at least two events) had the episodes of both the arterial and venous thromboembolism as, in detail, described in Table 1. In 65 patients (16 men and 49 women), the precipitating factors (surgery, injury, gravidity, drugs) were identified, in the rest of the patients the thromboembolic events were idiopathic. The used thrombophilic screening involved the following tests: activated partial thromboplastin time, prothrombin time, thrombin time, diluted Russell's viper venom time, kaolin clotting time, thromboplastin inhibition test, anticardiolipin antibodies, plasma level of fibrinogen, ProC Global test, activity and antigen levels of protein C, protein S and antithrombin, activity levels of factor VIII and factor FIX, plasma level of homocysteine, DNA testing for factor V Leiden, prothrombin G20210A and methylene tetrahydropholate reductase C677T. In addition, 156 blood donors without thrombosis and without family relation with the patients (86 men with mean age 43.3 ± 8.5 years and age range 26–67 years and 70 women with mean age 46.1 ± 9.5 years and age range 25–68 years) were tested as a control group. Genomic DNA was isolated from white blood cells according to the method described by Madisen et al. [9]. Genotyping was performed by polymerase chain reaction with subsequent restriction fragment length polymorphism analysis as previously described [4]. Restriction endonuclease Bsh1236I (MBI Fermentas) was used for the identification of the mutation. DNA fragments were separated by electrophoresis on a 9% polyacrylamide gel. The A allele gave one fragment with 208 bp, the G allele gave two fragments consisting of 190 and 18 bp. Of 400 subjects enrolled in the study, only one individual from the control group was a heterozygote for the A allele. No heterozygote for the A allele was found in the patient group, as well as none of the tested individuals was a homozygote for the A allele. As for the prevalence of the A allele, the results of our study are similar to the findings of other studies [1, 4, 7, 8]. We agreed with the above-cited authors that the 127G to A substitution is an infrequent finding. We did not identify the 127A allele in any patient suffering from thromboembolism and thus we were not able to testify the supposed relation between the examined mutation and thromboembolic events. However, even if such relation exists, because of its low prevalence the 127G to A mutation is likely to be the cause of thrombosis only in a limited number of patients. Of course, the size of the tested groups did not allow us to make a definitive conclusion. However, we agree that the 127A allele of the TM 127G/A polymorphism does not seem to be very frequent and thus it is not likely to be a common cause of thromboembolic events. We acknowledge the support of grant KEGA 3/1131/03.

The resistance to activated protein C (APC-resistance) based on the presence of factor V Leiden (F V Leiden) is the most frequent thrombophilic condition in the white race population. It contributes to the origin of thrombosis especially in the venous part of blood vessels. Significant geographic differences have been detected within Europe. The aim of this retrospective study was to determine the frequency in the occurrence of F V Leiden: 1. in healthy (asymptomatic) Slovak population, 2. in their consanguineously unrelated members with thrombosis and 3. in patients with myocardial infarction (IM) without or with other known risk factors of this disease (nicotinism, obesity, hypertension, dyslipoproteinemia, diabetes mellitus), respectively. The detection of FV Leiden was made by molecular biology methods. The occurrence in a group of 152 healthy individuals was four % (6 persons) and this frequency corresponds to the geographic localization of the Slovak Republic in Europe. In a group of 349 patients with thrombosis in anamnesis, FV Leiden was detected in 103 persons (29.5%). The occurrence was higher than the usually reported incidence in these patients (20%). Likewise, in a group of 35 patients with IM without risk factors in anamnesis, the occurrence of FV Leiden (8.6%) was significantly higher in comparison with healthy population and the incidence further increased significantly in a group of 41 patients with IM and the presence of at least one risk factor (14.6%). The authors therefore suppose an active role of the Leiden mutation of FV gene in the pathogenesis of this disease.