Publications by authors named "Ludek Slavik"

30 Publications

  • Page 1 of 1

A synergy of liquid chromatography with high-resolution mass spectrometry and coagulation test for determination of direct oral anticoagulants for clinical and toxicological purposes.

Biomed Chromatogr 2021 Oct 21;35(10):e5195. Epub 2021 Jun 21.

Department of Forensic Medicine and Medical Law, Faculty of Medicine and Dentistry, Palacký University Olomouc and University Hospital Olomouc, Olomouc, Czech Republic.

Direct oral anticoagulants are an alternative to anticoagulants based on vitamin K antagonists. Monitoring of direct oral anticoagulant concentration levels is necessary in specific cases (e.g. in emergency conditions, for determination of the cause of bleeding, adverse effects, risk of drug-direct oral anticoagulants interaction); therefore, a sensitive and specific method is needed. A methanol protein precipitation method followed by liquid chromatography with high-resolution mass spectrometry was developed for simultaneous separation and determination of apixaban, betrixaban, edoxaban, dabigatran, rivaroxaban and ximelagatran. The proposed method was fully validated in terms of linearity, the limits of detection and quantification, intra- and inter-day trueness and precision, recovery, matrix effect, process efficiency and stability. The method shows a strong correlation (Pearson's correlation coefficients > 0.92) with coagulation assays of apixaban, dabigatran and rivaroxaban (dilute thrombin time for gatrans and anti Xa factor (anti-Xa) activity for xabans). In addition, the developed method was applied for the identification and determination of apixaban and dabigatran in post-mortem serum samples. The developed method is a good alternative to coagulation tests which may show various interferences.
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http://dx.doi.org/10.1002/bmc.5195DOI Listing
October 2021

Hypertriglyceridemic Waist in Patients with Type 2 Diabetes: Its Relationship to Selected Markers of Vascular Damage.

Metab Syndr Relat Disord 2021 09 4;19(7):393-400. Epub 2021 Jun 4.

Department of Hemato-Oncology, University Hospital Olomouc and Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic, Olomouc, Czech Republic.

To evaluate the association between hypertriglyceridemic waist (HTGW), a promising marker of visceral adiposity and cardiovascular (CV) risk, and different indicators of vascular damage in type 2 diabetes (T2D) patients. This case-control study included 161 patients with T2D (91 males, 70 females) and 40 healthy controls (24 males, 16 females). HTWG was defined as waist circumference >90 cm in men or >85 cm in women and triglyceride concentrations >2 mmol/L. In addition to anthropometric and metabolic parameters, markers of endothelial dysfunction, namely von Willebrand factor (vWF) and plasminogen activator inhibitor-1 (PAI-1), were assessed. Arterial stiffness parameters were examined using the SphygmoCor system. Individuals with T2D and HTGW showed the highest elevation of PAI-1 levels and significantly increased vWF levels compared with healthy controls. No significant differences in arterial stiffness markers were observed between T2D individuals. Age and, for several markers, systolic and/or diastolic blood pressure were identified as the main predictors for arterial stiffness, whereas PAI-1 and vWF levels were predicted by metabolic parameters. HTGW represents increased CV risk in T2D patients, mainly due to endothelial damage. The presence of HTGW had no significant effect on arterial stiffness compared with other T2D individuals.
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http://dx.doi.org/10.1089/met.2021.0019DOI Listing
September 2021

Use of Fibrinogen Determination Methods in Differential Diagnosis of Hypofibrinogenemia and Dysfibrinogenemia.

Clin Lab 2021 Apr;67(4)

Background: Fibrinogen plays an important role in hemostasis. The normal concentration of fibrinogen in blood plasma is between 1.8 - 4.2 g/L. Decreased fibrinogen levels are observed in congenital afibrinogenemia, hypofibrinogenemia, dysfibrinogenemia, disseminated intravascular coagulation, fibrinolytic therapy, some more severe hepatic parenchymal disorders, and increased blood loss. Elevated fibrinogen levels occur in inflammatory diseases and neoplastic diseases, in pregnancy, and postoperative conditions. Functional fibrinogen measurement is also one of the basic coagulation screening tests. The fibrinogen antigen assay is used to distinguish between qualitative and quantitative fibrinogen disorders.

Methods: The aim of the study was the use of fibrinogen determination methods in differential diagnosis of hypofibrinogenemia and dysfibrinogenemia, statistical evaluation and determine the relationship of fibrinogen Clauss assay, prothrombin time (PT) derived fibrinogen assay, and fibrinogen antigen in the group of 60 patients with congenital fibrinogen disorders (n = 40 dysfibrinogenemia; n = 20 hypofibrinogenemia).

Results: The results measured by the PT-derived fibrinogen assay were approximately four times higher compared to the fibrinogen Clauss assay in the group of patients with dysfibrinogenemia. In patients with hypofibrinogenemia, there is a correlation (r = 0.9016) between the fibrinogen Clauss assay and PT-derived fibrinogen assay with a statistical significance of p < 0.0001. Using a linear or quadratic interpolation function, we were able to determine the fibrinogen Clauss assay and the fibrinogen antigen assay before analysis.

Conclusions: The higher level of the PT-derived fibrinogen assay compared to the fibrinogen Clauss assay in the group of patients with dysfibrinogenemia may pose a greater risk to asymptomatic patients who require diagnosis and treatment in case of bleeding. The fibrinogen value using the PT-derived fibrinogen assay could erroneously give a normal level. The use of the interpolation function is important to estimate the value of fibrinogen activity and antigen before the analysis itself by the Clauss assay or analysis by the fibrinogen antigen assay.
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http://dx.doi.org/10.7754/Clin.Lab.2020.200820DOI Listing
April 2021

Current Promising Biomarkers and Methods in the Diagnostics of Antiphospholipid Syndrome: A Review.

Biomedicines 2021 Feb 8;9(2). Epub 2021 Feb 8.

Department of Immunology, Faculty of Medicine and Dentistry, Palacky University Olomouc and Faculty Hospital, 775 15 Olomouc, Czech Republic.

Antiphospholipid syndrome (APS) is a hypercoagulation condition associated with the incidence of heterogenic antiphospholipid antibodies (aPLs), which non-specifically affect hemostasis processes. APS is clinically manifested by recurrent arterial and venous thromboses and reproduction losses. The aPL antibodies, which may induce clinical manifestations of APS, include criteria antibodies anti-cardiolipin, anti-β2-glycoprotein-I, and lupus anticoagulant, but also non-criteria antibodies, for example anti-β2-glycoprotein-I domain I, anti-phosphatidylserine/prothrombin, anti-annexin V, and many others. APS occurs mostly in patients of younger and middle age, most frequently in females. Laboratory diagnostics of APS are quite difficult, as they include a wide spectrum of examining methods, which are based on various principles of detection and are performed using various laboratory techniques. The objective of the review is to describe the current state of potentially examined biomarkers and methods in APS diagnostics. The aforementioned biomarkers are lupus anticoagulant, anti-β2-glycoprotein-I, anti-cardiolipin, anti-β2-glycoprotein-I domain I, anti-phosphatidylserine/prothrombin, anti-β2-glycoprotein-I IgA, anti-cardiolipin IgA, anti-annexin V and II, anti-prothrombin, anti-cardiolipin/vimentin, anti-protein S/protein C, and antibodies against phospholipid antigens for whose diagnostics we may use some of the methods established for a long time and some of the modern methods-the coagulation method for the determination of lupus anticoagulant (LA), enzyme-linked imunosorbent assay (ELISA), chemiluminescence analysis (CLIA), multiplex fluorescence flow immunoassay (MFFIA), fluorescence enzyme immunoassay (EliA), line immunoassay (LIA), multiline dot assay (MLDA), and thin-layer chromatography (TLC). Conclusion: Antibodies against phosphatidylethanolamine, phosphatidic acid, phosphatidylserine, phosphatidylinositol, cardiolipin/vimentin complex, and annexin V are currently the most studied new markers. However, these assays have not been standardized until now, both from the laboratory and clinical point of view. In this review we summarize the evidence of the most studied aPL markers and their potential clinical significance in seronegative APS (SN-APS).
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http://dx.doi.org/10.3390/biomedicines9020166DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7914732PMC
February 2021

The Modern Pneumatic Tube System Transports with Reduced Speed Does Not Affect Special Coagulation Tests.

J Med Syst 2020 Jul 21;44(9):142. Epub 2020 Jul 21.

Department of Blood Transfusion, University Hospital Olomouc, Olomouc, Czech Republic.

Pneumatic tube transport systems (PTS) for delivery of patient samples to a hemostasis laboratory are often used to reduce turnaround time for vital analyses. PTS in our hospital has the ability to regulate the transport speed in the range of 3-6 m/s with acceleration control technology. We evaluated the effects of PTS transport for routine coagulation tests, platelet function tests and special global coagulation tests. Duplicate samples were collected from 29 patients and 40 healthy individuals. One sample was sent using PTS and the other was carried by personnel to the lab for determination of protrombin time, activated partial thromboplastin time, trombin time, fibrinogen, antitrombin and thrombin generation test. Platelet function was measured by means of a Apact 4004® analyzer using the inductors (ADP, Arachidonic acid and Epinephrine). Samples transported using PTS with normal transport speed 6 m/s does not affect basic coagulation tests (PT, aPTT, FIB, TT and AT), but TGT has significantly altered. The use of PTS with controlled acceleration regulated the increase in thrombin generation from 10% to 3%, which is not statistically signifiant. The use of PTS with controlled acceleration did not show a significant difference even with the highly sensitive method of platelet aggregation. We conclude that PTS with acceleration control with transport speed from 3 to 6 m/s does not affect to platelet activity as measured by LTA and also global coagulation test - TGT. The advantage of PTS transport is very rapid assessment laboratory testing. From the above validation study, it is clear that PTS should always be validated for specialized laboratory methods and appropriately adapted to specific transport conditions.
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http://dx.doi.org/10.1007/s10916-020-01614-6DOI Listing
July 2020

17β-Estradiol Promotes Proinflammatory and Procoagulatory Phenotype of Innate Immune Cells in the Presence of Antiphospholipid Antibodies.

Biomedicines 2020 Jun 15;8(6). Epub 2020 Jun 15.

Department of Immunology, Faculty of Medicine and Dentistry, Palacky University Olomouc and Faculty Hospital, 775 15 Olomouc, Czech Republic.

Antiphospholipid syndrome (APS) is the most common cause of acquired thrombophilia and recurrent spontaneous miscarriages associated with extended persistence of antiphospholipid antibodies (aPL). How circulating aPL and high-17β-estradiol (E2) environment contribute to the pregnancy complications in APS is poorly defined. Therefore, we aimed to analyse whether E2 could be responsible for the immune cell hyperactivation in aPL- positive (lupus anticoagulant, anti-cardiolipin, anti-β2-glycoprotein) in women. For this, peripheral blood mononuclear cells (PBMCs) from 14 aPL- positive and 13 aPL- negative women were cultured in the presence or absence of E2, LPS or E2+LPS and cell immunophenotype and cytokine release were analysed. In the aPL+ group, E2 presence markedly increased the percentage of NK cells positive for CD69 ( < 0.05), monocytes positive for tissue factor (TF, CD142) ( < 0.05), and B cells expressing PD-L1 ( < 0.05), as well as the elevated production of IL-1β comparing to aPL- women ( < 0.01). Regardless of aPL positivity, E2 augmented the procoagulatory response elicited by LPS in monocytes. Our findings show the ability of E2 to promote proinflammatory and procoagulatory phenotype of innate immune cells in individuals with aPL positivity. Our data highlights the significant impact of female hormones on the activation of immune cells in the presence of aPL.
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http://dx.doi.org/10.3390/biomedicines8060162DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7345022PMC
June 2020

Anti-domain 1 β2 glycoprotein antibodies increase expression of tissue factor on monocytes and activate NK Cells and CD8+ cells in vitro.

Auto Immun Highlights 2020 Mar 2;11(1). Epub 2020 Mar 2.

Department of Immunology, Faculty of Medicine and Dentistry, Palacky University Olomouc and Faculty Hospital, Olomouc, Czech Republic.

Background: β2-Glycoprotein I (β2GPI) represents the major antigenic target for antiphospholipid antibodies (aPL), with domain 1 (D1) being identified as a risk factor for thrombosis and pregnancy complications in APS. We aimed to analyse the ability of aPL, and particularly anti-D1 β2GPI, to stimulate prothrombotic and proinflammatory activity of immune cells in vitro.

Methods: Peripheral blood mononuclear cells (PBMCs) from 11 healthy individuals were incubated with: (1) "anti-D1(+)"-pooled plasma derived from patients suspected of having APS contained anticardiolipin antibodies (aCL), lupus anticoagulant (LA), anti-β2GPI and anti-D1 β2GPI; (2) "anti-D1(-)"-pooled plasma from patients suspected of having APS contained aCL, LA, anti-β2GPI, and negative for anti-D1 β2GPI; (3) "seronegative"-negative for aPL.

Results: The presence of anti-D1(+) and anti-D1(-) plasma resulted in increased HLA-DR and CD11b on monocytes. While only anti-D1(+) plasma markedly increased the percentage and median fluorescence intensity (MFI) of CD142 (tissue factor, TF) on monocytes in comparison with those cultured with anti-D1(-) and seronegative plasma. Anti-D1(+) plasma resulted in increased percentage and MFI of activation marker CD69 on NK and T cytotoxic cells. Expression of IgG receptor FcγRIII(CD16) on monocytes and NK cells was down-regulated by the anti-D1(+) plasma.

Conclusions: Taking together, our study shows the ability of patient-derived aPL to induce immune cell activation and TF expression on monocytes. For the first time, we demonstrated the influence of anti-D1 β2GPI on the activation status of monocytes, NK and cytotoxic T cells. Our findings further support a crucial role of D1 epitope in the promotion of thrombosis and obstetrical complications in APS.
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http://dx.doi.org/10.1186/s13317-020-00128-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7065342PMC
March 2020

Monitoring of fibrinolytic system activity with plasminogen, D-dimers and FDP in primary total knee arthroplasty (TKA) after topical, intravenous or combined administration of tranexamic acid.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2020 Jun 16;164(2):168-176. Epub 2019 Sep 16.

Department of Orthopaedics, University Hospital Olomouc and Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic.

Aim: We assessed various ways of tranexamic acid (TXA) administration on the fibrinolytic system. Blood loss, transfusions, drainage and haematoma were secondary outcomes.

Methods: In this prospective study, we examined 100 patients undergoing primary total knee arthroplasty (TKA) between June and November 2018. Patients were randomly assigned to 4 groups according to the following TXA regimens: 1) loading dose 15 mg TXA/kg single intravenous administration applied at initiation of anesthesia (IV1); 2) loading dose 15 mg TXA/kg + additional dose 15 mg TXA/kg 6 h after the first application of TXA (IV2); 3) IV1 regime in combination with a local wash of 2 g of TXA in 50 mL of saline (COMB); 4) topical administration of 2 g of TXA in 50 mL of saline (TOP).

Results: Systemic fibrinolysis interference was insignificant in all of the regimens; we did not detect significant differences between IV1, IV2 and COMB in the monitored parameters within the elapsed time after the TKA; IV regimes had the lowest total drainage blood loss; the lowest blood loss was associated with the IV1 and IV2 regimens (IV1, IV2 < COMB < TOP); the lowest incidence of haematomas was in patients treated with TXA topically (i.e., in COMB + TOP).

Conclusion: The largest antifibrinolytic effect was associated with intravenous administration of TXA. In terms of blood loss, intravenously administered TXA can interfere with the processes associated with the formation of the fibrin plug more efficiently than the simple washing of wound surfaces with TXA.
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http://dx.doi.org/10.5507/bp.2019.034DOI Listing
June 2020

Searching for genetic variants associated with thrombophilia.

Cas Lek Cesk 2019 ;158(1):28-32

Thrombotic states are inherited or acquired predisposition for thrombosis in the human vascular system. Nowadays Leiden mutation and mutation in prothrombin G20210A contributing to congenital thrombophilia are routinely tested. These mutations have a high prevalence in the population. Congenital deficiencies of protein S, protein C and antithrombin III are rare thrombophilia with lower population frequency, but higher risk of thromboembolic event. The genetic causes are mutations in the genes, which encode these proteins. The choice of proper molecular genetic testing depends on the difference in the detection of well-known single nucleotide polymorphism or unknown/rare variant. For the detection of causative variant FV Leiden and prothrombin G20210A are mostly used PCR-RFLP, reverse Strip Assay®, allele-specific PCR, TaqMan real-time PCR and SNaPshot®. Precise patient selection should precede the genetic testing of rare variants in anticoagulant proteins. It is appropriate to use methodology of massive parallel sequencing supplemented by a methodology for the detection of larger gene rearrangements - MLPA. We are successfully employing this approach in our institute. This methodology is faster with larger analytic capacity compared to commonly used direct sequencing by Sanger method.
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June 2019

Multianalyte Determination of NOACs Using LC-MS/MS and Comparison with Functional Coagulation Assays.

Clin Lab 2018 Oct;64(10):1611-1621

Background: Detection of new oral anticoagulant (NOAC) levels by screening, special and global tests, and liquid chromatography-coupled tandem mass spectrometry (LC-MS/MS) is important in clinical situations when the cause of bleeding needs to be determined.

Methods: We compared a routine coagulation test, special function test for NOACs, global coagulation test, and an LC-MS/MS method that enables simultaneous determination of apixaban, dabigatran and rivaroxaban in human plasma within one analysis to determine the optimal indication of the comparison methods, including their limitations and interferences.

Results: This study was conducted on a set of blood samples from 116 patients treated with NOACs. The results of both specific dilute thrombin time (dTT) tests for dabigatran provided the same results as the activated partial thromboplastin time (aPTT) screening test in comparison with LC-MS/MS as a reference. The dTT assay HemosIL® showed better results for low concentrations when compared to LC-MS/MS than dTT HYPHEN® as HemosIL® uses a non-linear calibration curve. Results of the specific anti-Xa assay yielded better results than the prothrombin time test in comparison with LC-MS/MS as a reference, especially for apixaban, but also for rivaroxaban. Our LC MS/MS method is simply feasible, but only in a specialized laboratory. The method is easy-to-use for the simultaneous determination of all dabigatran, apixaban and rivaroxaban by LC-MS/MS within three minutes with a concentration range of 1 to 500 µg/L without dilution.

Conclusions: In the normal practice of the coagulation laboratory, it is advisable to use specific tests for NOAC determination as screening and global assays are not sufficiently specific. The dTT test is the optimal choice for dabigatran determination and for xabans to determine anti-Xa activity. The LC-MS/MS method is suitable as an arbitration method for serious conditions.
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http://dx.doi.org/10.7754/Clin.Lab.2018.180335DOI Listing
October 2018

Possibility of Coagulation System Activation Determination with Tissue Factor in Pregnancy Complications.

Clin Lab 2016 Oct;62(10):1851-1856

Background: In this part of the study, where we determined the causes of preeclampsia and other obstetric complications, we focused on the role of tissue factor (TF) in the activation of these pathophysiological processes. Recent findings attribute a significant part of the activation of coagulation creation of autoantibodies. Once this mechanism is activated, the antibodies induce expression of tissue factor (TF, CD142) on monocytes and vascular endothelial cells.

Methods: We have proposed a monitor activation model of the coagulation system in preeclampsia and other pregnancy complications using TF expression on monocytes by flow cytometry and simultaneous determination the TF-induced thrombin generation in plasma. To determine expression of tissue factor (CD142) on monocytes, we proposed a method of multicolor flow cytometry using anti CD45 PerCP, anti CD14 APC, anti CD16b FITC, and anti CD142 PE antibodies and the corresponding isotype controls.

Results: We verified the model on patients with severe antiphospholipid syndrome, which is a high expression of antibodies, in particular against beta-2GPI.

Conclusions: We demonstrated complete inhibition of TF expression on monocytes and a significant reduction of thrombin generation in plasma.
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http://dx.doi.org/10.7754/Clin.Lab.2016.151234DOI Listing
October 2016

Polymorphism of the Fcγ Receptor II as a Possible Predisposing Factor for Heparin-Induced Thrombocytopenia.

Clin Lab 2015 ;61(8):1027-32

Background: Heparin-induced thrombocytopenia (HIT) represents a serious complication of heparin treatment. IgG antibodies binding platelet factor 4 (PF4) and heparin trigger the clinical manifestations of HIT. However, only a portion of the antibodies have the ability to activate platelets, and these can be identified by a platelet aggregation test (functional testing). However, this expression has been detected to have a molecular cause, which is a mutation of FcγRIIa. The FcγRIIa receptor is responsible for the activation of platelets by antibodies in HIT.

Methods: To determine HIT, impedance aggregometry using the Multiplate analyzer (MEA) as heparin-induced aggregation technique and the Technozym HIT IgG ELISA test were used. The MEA method uses sensitization of donor platelets with patient plasma in the presence of heparin at a concentration of 0.5 IU/mL. The results were compared with the ELISA test. Mutation of FcγRHa was assessed using the asymmetric real-time PCR method that is based on the reaction with two hybridization probes and melting curve analysis.

Results: Examined were 100 patients at a clinically intermediate and higher risk of HIT according to the 4T's score. All samples were examined by the ELISA test and MEA, with positive samples being further confirmed by high-concentration heparin. In the group of patients, 10.0% were positive by MEA as compared with 4% determined by ELISA. The results of genetic analysis of FcγRIIa did not provide statistically significant differences between positive patients found by the functional test as well as the ELISA test and seronegative patients.

Conclusions: The genetic mutation FcγRIIa is a predisposing factor for manifestation of HIT in the form of thrombocytopenia, but the process of seroconversion apparently needs another inducing factor. Therefore, the examination of mutations can be classified as predisposing factors rather than to confirm the diagnosis of HIT.
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http://dx.doi.org/10.7754/clin.lab.2015.141207DOI Listing
October 2015

Markers of endothelial activation in preeclampsia.

Clin Lab 2015 ;61(1-2):39-46

Background: The study aimed at finding a laboratory approach to detect endothelial damage in normal pregnancy as well as in pregnancy complicated by preeclampsia using selected markers of endothelial activation.

Materials: A total of 403 healthy pregnant women without a history of deep vein thrombosis and/or hypertension were prospectively studied. From all women, venous blood was collected before the end of the 1st trimester, between weeks 24 and 28 of gestation, and in the 3rd trimester (weeks 34-36). Assays of tissue plasminogen activator, plasminogen activator inhibitor-1, von Willebrand factor activity and antigen, thrombomodulin, endothelial protein C receptor, and endothelial microparticles activated by TF were performed.

Results: When comparing women who developed preeclampsia during pregnancy (the average levels were 23.41 μg/L, 34.33 μg/L, and 53.56 μg/L in the 1st, 2nd, and 3rd trimesters, respectively) with healthy pregnant women (the average levels were 19.05 μg/L, 28.47 μg/L, and 39.86 μg/L in the 1st, 2nd, and 3rd trimesters, respectively) significant differences in the levels of thrombomodulin were found in all three trimesters. By contrast, no statistically significant differences in the levels of vWF (both antigen and activity), t-PA, EPCR, EMPs, MMP-2, MMP-9, and TIMP-9 were found in any trimesters in the same group.

Conclusions: Pregnancy and preeclampsia strongly influence the levels of studied markers. The findings of this work confirm the possible predictive potential of thrombomodulin and PA-1.
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April 2015

The role of tissue factor in normal pregnancy and in the development of preeclampsia: A review.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2015 Jun 22;159(2):192-6. Epub 2014 Dec 22.

Department of Obstetrics and Gynecology, University Hospital Olomouc and Faculty of Medicine and Dentistry, Palacky University Olomouc.

Background: Tissue factor (TF) is a key element for normal gestation, especially in the first trimester. TF levels are hence raised in pregnancy, producing an adaptive hypercoagulable state. Potentiated hypercoagulability however, is associated with disorders of pregnancy such as pre-eclampsia but the results of TF and its inhibitor, tissue factor pathway inhibitor (TFPI), measurement, in pre eclampsic women are ambiguous and the data conflicting. This review covers the current knowledge status of the role of TF assessment in pregnancy with a focus on its diagnostic utility.

Methods: A review of the literature using the following key words: tissue factor, thrombosis, inflammation, pregnancy, preeclampsia.

Results: The published literature shows raised and unchanged TF levels in various studies of pre-eclampsia along with equally conflicting data for TFPI. The various study designs and methods used in these studies makes valid comparison difficult. Meta analysis of 34 randomized trials showed that low-dose aspirin in early phases of gravidity (starting from the 16th week or earlier) significantly reduces the incidence of preeclampsia.

Conclusions: Overall, the results of the literature search together with knowledge of the structure and biological effects of TF, suggest that measuring the level of plasma TF/TFPI is not ideal for determining the actual levels of TF in the uteroplacental circulation. The current view that endothelial dysfunction is the trigger for preeclampsia, suggests that aspirin may be an effective prophylaxis. Further research will be necessary: measuring the expression of tissue factor on monocytes using flowcytometry and comparing the development of this expression during normal pregnancy and pregnancy complicated by preeclampsia, for example. Another possibility is immunohistochemical determination of the level of TF expression directly in placental tissue.
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http://dx.doi.org/10.5507/bp.2014.061DOI Listing
June 2015

Genetic polymorphisms of platelet receptors in patients with acute myocardial infarction and resistance to antiplatelet therapy.

Genet Test Mol Biomarkers 2014 Sep 5;18(9):599-604. Epub 2014 Aug 5.

1 Department of Hemato-Oncology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital , Olomouc, Czech Republic .

Methods: The studied group comprises 124 patients with acute myocardial infarction on dual antiplatelet therapy with acetylsalicylic acid (ASA) and thienopyridines. Antiplatelet therapy was monitored by platelet-rich plasma light transmittance aggregometry (LTA) using the APACT 4004 analyzer (Helena Laboratories) and by whole blood impedance aggregometry (multiple electrode aggregometry [MEA]) using the Multiplate analyzer (Dynabyte). Platelet aggregation was detected after stimulation with arachidonic acid for detection of aspirin resistance and with adenosine diphosphate (ADP) and prostaglandin E1 for detection of thienopyridine resistance. To determine the frequencies of P2Y12 (i-744T>C; rs2046934), P2Y12 (34C>T; rs6785930), COX-1 (-842A>G; rs10306114), GPVI (13254T>C; rs1613662), and GPIbA (5T>C; rs2243093) polymorphisms, DNA of patients with AIM was tested by real-time-polymerase chain reaction and melting curve analysis using the LightCycler 480 analyzer (Roche Diagnostics).

Results: The cut-off points used for patients with effective ASA therapy are 25% of aggregated platelets and 220 area under the curve (AUC)/min if LTA or MEA, respectively. The cut-off points used for effective thienopyridine therapy are 45% of aggregated platelets or 298 AUC/min, respectively. Both LTA and MEA found that aspirin and thienopyridine therapies failed in 14.51% and 25.8%, respectively. The data were statistically processed using the SPSS version 15 software (SPSS, Inc.). Associations between receptor mutation status and response to therapy were assessed with Fisher's exact test. The significance level was set at 0.05.

Conclusion: The aim of our work was to use the two functional laboratory methods described earlier to assess both aspirin and thienopyridine resistance and to determine the contribution of genetic polymorphisms of platelet receptors to resistance to antiplatelet therapy in AIM. Fisher's exact test showed a significant statistical correlation between platelet function tests suitable for monitoring ASA resistance, that is, LTA and MEA, and mutation status of COX1_A1 (-A842G). Fisher's exact test showed no statistically significant correlations between platelet function tests suitable for monitoring ASA resistance, that is, LTA and MEA, and mutation status of GP1bA (-5T>C) and GP6 (T13254C). Fisher's exact test showed no statistically significant correlation between mutational statuses of the receptors P2RY12 (i-T744C), P2RY12 (C34T), GP1bA (-5T>C), or GP6 (T13254C) and response to antiplatelet therapy with 75 mg of clopidogrel.
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http://dx.doi.org/10.1089/gtmb.2014.0077DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4150366PMC
September 2014

The influence of apolipoprotein A5 T-1131C and apolipoprotein E common genetic variants on the levels of hemostatic markers in dyslipidemic patients.

Clin Chim Acta 2014 Sep 9;436:11-7. Epub 2014 May 9.

Department of Haemato-Oncology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc, I.P. Pavlova 6, 775 20 Olomouc, Czech Republic.

Objectives: The aim of this study was to evaluate the relationships of the T-1131C (rs662799) polymorphism variants of apolipoprotein A5 (Apo A5) gene and variants of apolipoprotein E (Apo E) gene common polymorphism (rs429358, rs7412) to selected hemostatic markers.

Study Design And Methods: We examined 590 asymptomatic dyslipidemic patients, subsequently divided into MetS+ (n=146) and MetS- (n=444) groups according to the criteria for identification of the metabolic syndrome (MetS). We compared variant frequencies and differences in levels of hemostatic markers according to Apo A5, Apo E and Apo A5/Apo E common variants.

Results: The -1131C Apo A5 minor variant was associated with elevated tissue plasminogen activator (tPA) in comparison to TT genotype (p<0.001), but not in the MetS+ group. The analysis of Apo A5/Apo E common variants in all subjects revealed that the presence of -1131C minor allele has always been associated with higher levels of tPA in comparison with T allele, regardless of Apo E genotype. Also the presence of minor Apo E2 allele led to elevated tPA concentrations in both T and C carriers. In addition, common -1131C/E2 variant was associated with the highest tPA levels.

Conclusion: We demonstrated a remarkable association especially between the -1131C Apo A5 variant and increased tPA levels in asymptomatic dyslipidemic patients.
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http://dx.doi.org/10.1016/j.cca.2014.04.029DOI Listing
September 2014

Positive association of adiponectin with soluble thrombomodulin, von Willebrand factor and soluble VCAM-1 in dyslipidemic subjects.

Clin Biochem 2013 Jun 13;46(9):766-71. Epub 2013 Mar 13.

3rd Department of Internal Medicine, Faculty of Medicine and Dentistry UP Olomouc and University Hospital Olomouc, Czech Republic.

Objectives: Both decreased and increased risk of cardiovascular events/mortality have been reported with high adiponectin levels. Only a few studies have reported an association of adiponectin with markers of hemostasis/endothelial dysfunction which might explain the reported discrepancies.

Design And Methods: We evaluated the association of total adiponectin with von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1), tissue plasminogen activator (t-PA), soluble thrombomodulin (sTM), adhesion molecules sICAM-1 and sVCAM-1, lipids and markers of insulin resistance (IR) in 308 asymptomatic dyslipidemic subjects and healthy controls. Subjects were divided into 4 dyslipidemic phenotypes (DLP): DLP1 (TG < 1.5 mmol/L + ApoB < 1.2 g/L), DLP2 (TG ≥ 1.5 + ApoB < 1.2), DLP3 (TG < 1.5 + ApoB ≥ 1.2) and DLP4 (TG ≥ 1.5 + ApoB ≥ 1.2). The results were evaluated also according to the presence (+) and absence (-) of metabolic syndrome (MS).

Results: In hyperlipidemic subjects (DLP2-4), PAI-1, t-PA and sICAM-1 correlated with markers of IR but only t-PA correlated inversely with adiponectin. In contrast positive association of adiponectin with vWF, sTM and sVCAM-1 was found but none of these parameters correlated with markers of insulin resistance. In multiple regression analysis, adiponectin remained independently associated with vWF [in DLP3, DLP4, DLP2-4, MS(-)], with sTM [in DLP2, DLP4, DLP2-4, MS(+)] and with sVCAM-1 [in DLP2, DLP3, DLP4, DLP2-4, MS(+)]. In healthy controls (DLP1), no association between adiponectin and markers of hemostasis/endothelial dysfunction was found.

Conclusion: The independent positive association of adiponectin with vWF, sTM and sVCAM-1 deserves further evaluation in connection with the risk of atherothrombotic cardiovascular events.
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http://dx.doi.org/10.1016/j.clinbiochem.2013.02.014DOI Listing
June 2013

Relationship between serum adipocyte fatty acid-binding protein and endothelial/hemostatic markers in dyslipidemic subjects.

Neuro Endocrinol Lett 2012 ;33 Suppl 2:26-31

3rd Department of Internal Medicine, University Hospital Olomouc, Czech Republic.

Objectives: Some findings support the role of serum adipocyte fatty acid-binding protein (A-FABP) as a key pro-inflammatory mediator that links obesity with cardiovascular diseases. The aim of the study was to evaluate the association of A-FABP with endothelial/hemostatic markers [von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1), tissue-plasminogen activator (t-PA), soluble intercellular cell adhesion molecule-1 (s-ICAM-1) and soluble vascular cell adhesion molecule-1 (s-VCAM-1)] in asymptomatic dyslipidemic subjects.

Design: We examined 105 dyslipidemic patients (with apolipoprotein B concentration ≥1.2 g/l and/or triglyceride (TG) concentration ≥1.5 mmol/l) without clinical manifestation of atherosclerosis and 50 normolipidemic healthy subjects, who served as a control group. Except of endothelial/hemostatic markers, anthropometric and lipid parameters, markers of insulin resistance and inflammation were assessed.

Results: In dyslipidemic patients, A-FABP positively correlated with age (p<0.05), TG (p<0.05), insulin (p<0.05), homeostatic model assessment (HOMA) index (p<0.05), body mass index (p<0.001), waist circumference (p<0.05), high sensitivity C reactive protein (p<0.01), and vWF (p<0.05) and negatively with male gender (p<0.05). There were no correlations between A-FABP and PAI-1, t-PA, s-VCAM-1 or s-ICAM-1. By using linear multivariate regression analysis the positive association between A-FABP and vWF was independent of age, gender, insulin resistance, and visceral obesity.

Conclusion: Study displayed an independent positive association of A-FABP with vWF in clinically asymptomatic dyslipidemic subjects. Contribution of A-FABP in the process of endothelial dysfunction could help to explain the role of obesity in cardiovascular damage.
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April 2013

Assessment of relationship between acute ischemic stroke and heart disease--protocol of a prospective observational trial.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2012 Sep 4;156(3):284-9. Epub 2012 Oct 4.

Comprehensive Stroke Center, Department of Neurology, University Hospital Olomouc and Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic.

Background: Stroke and acute myocardial infarction are the leading causes of death and disability in industrialized countries. Multiple interactions exist between the various forms of cardiovascular and cerebrovascular diseases, and risk factors for development of stroke and major cardiovascular events are similar. There is currently no clear link between acute coronary syndrome and stroke, although it has been repeatedly described. In addition, there are currently no clear recommendations for how to proceed in the case of signs of myocardial damage in patients with acute stroke and how to manage the next follow-up. METHODS-DESIGN: In this prospective observational trial, 500 consecutive ischemic stroke patients admitted at the Comprehensive Stroke Center will be enrolled within 12 h from stroke onset. The set of examinations will consist of: 1) Acute brain computed tomography or magnetic resonance imaging 2) Laboratory tests: A) within 12 h from stroke onset: NT pro B-type of natriuretic peptide, pro-atrial natriuretic peptide, creatinekinase MB, troponin T (cTnT), interleukin 6, procalcitonin, high sensitive C-reactive protein and D-dimers. B) control level of cTnT after 4 h from admission C) non-acute laboratory samples within 60 h from stroke onset: glycated haemoglobine, serum lipids; 3) Electrocardiogram (ECG) on admission and 4 h from stroke onset; 4) Transesophageal or transthoracal echocardiography and 24-h ECG-Holter within 15 days from stroke onset; 5) Neurosonological examination within 60 h from stroke onset; 6) Thirty patients with a positive finding of acute myocardial ischemia (ECG, cTnT) will be examined by coronary angiography (CAG); 7) Epidemiological data will be acquired.

Statistics: The epidemiological characteristics of the whole sample of patients; correlation between differences between group of cardioembolic ischemic stroke patients and group of patients with ischemic stroke of another etiology; correlation of infarction volume on DWI-MRI with the level of cTnT; correlation of the ECG findings with the level of cTnT and clinical signs; correlation of the CAG findings with level of cTnT and ECG findings will be statistically evaluated at the 5% level of statistical significance.

Conclusion: The main goal of the project is to improve identification of patients with acute coronary syndrome and with concurrent acute ischemic stroke as these patients require specific treatment and secondary prevention of ischemic events.

Trial Registration: Clinicaltrials.gov NCT01541163.
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http://dx.doi.org/10.5507/bp.2012.094DOI Listing
September 2012

The pathophysiology of endothelial function in pregnancy and the usefulness of endothelial markers.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2011 Dec;155(4):333-7

Department of Hemato-oncology, University Hospital Olomouc, Czech Republic.

Aim: The aim of this study was to assess coagulation markers of endothelial damage and examine new markers of endothelial activation such as matrix metalloproteinases (MMPs) in a group of healthy pregnant women. Matrix metalloproteinase (MMP)-2, in particular, plays a major role in the degradation of the extracellular matrix confirming its essential function in both the survival (angiogenesis) and death of endothelial cells. Detection of specific coagulation factors, mainly released from the vascular endothelium such as vWF, sTM (soluble thrombomodulin) and ePCR (endothelial protein C receptor) and factors dependent on endothelial activation such as t-PA and PAI-1, could provide information on possible endothelial dysfunction and help differentiate pregnant patients with an altered thrombotic state.

Methods: Healthy pregnant women underwent complete assessment for endothelial damage (as vWF, vWF activity, sTM, ePCR, EMP, MMP-2, MMP-9 and TIMP-2) using the ELISA and other methods.

Results And Conclusions: The results show that endothelial activation during pregnancy is different from that in other pathological conditions involving endothelial damage and typically characterized by higher levels of both coagulation endothelial markers and MMPs. In pregnancy, changes in extracellular matrix composition and matrix metalloproteinase activity also occur and promote vascular remodeling but, only in the uterus. Predisposing risk factors for epithelial dysfunction, and vascular mediators associated with vascular remodeling must be assessed from concentrations in whole blood. The levels of MMPs are not increased in the circulation and the local situation in the uterus cannot be monitored this way. However, MMP-2 processes and modulates the functions of many other vasoactive and pro-inflammatory molecules including adrenomedullin, big endothelin-1, calcitonin gene-related peptide, CCL7/MCP-3, CXCL12/SDF-1, galectin-3, IGFBP-3, IL-1 Beta, S100A8, and S100A9. These molecules represent new potential molecular markers of endothelial damage during pregnancy.
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http://dx.doi.org/10.5507/bp.2011.031DOI Listing
December 2011

Prothrombotic markers in asymptomatic dyslipidemic subjects.

J Thromb Thrombolysis 2011 Jan;31(1):27-36

3rd Department of Internal Medicine, University Hospital, IP Pavlova 6, 77520 Olomouc, Czech Republic.

The aim of this study was to evaluate the plasma levels of prothrombotic markers--von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1), tissue plasminogen activator (t-PA)--in asymptomatic subjects with dyslipidemia. Asymptomatic subjects with dyslipidemia and their relatives (n = 234) were assessed for lipids and prothrombotic markers. Individuals were divided into four dyslipidemic phenotypes (DLP) according to apolipoprotein B (apoB) and triglycerides (TG): DLP1 (n = 58, apoB < 1.2 g/l and TG < 1.5 mmol/l), DLP2 (n = 47, apoB < 1.2 g/l and TG ≥ 1.5 mmol/l), DLP3 (n = 31, apoB ≥ 1.2 g/l and TG < 1.5 mmol/l) and DLP4 (n = 98, apoB ≥ 1.2 g/l and TG ≥ 1.5 mmol/l). Associations between prothrombotic markers and risk factors for atherosclerosis, markers of insulin resistance, and the intima-media thickness of the common carotid artery (IMT) were assessed too. Significant differences in PAI-1 between normolipidemic phenotype--DLP1 (62.5 (35.9-82.9) ng/ml) and hypertriglyceridemic phenotypes--DLP2 (82.2 (61.1-122.1) ng/ml, p < 0.01) and DLP4 (91.4 (63.5-111.8) ng/ml, p < 0.001) after adjustment for age, sex and body mass index, were found. Levels of t-PA were different only between DLP1 and DLP4 (1.9 (0.9-3.3) ng/ml vs. 5.3 (2.5-8.6) ng/ml, p < 0.05). There were no significant differences of vWF between DLPs. PAI-1 and t-PA correlated with lipid parameters, markers of insulin resistance, blood pressure and obesity. VWF was independently associated with IMT, which was increased in DLP4. Individuals with hypertriglyceridemic phenotypes showed increased levels of PAI-1 in comparison with normolipidemic subjects. The elevation of t-PA was presented only in patients with simultaneously elevated TG and apoB. The significant increase of IMT confirmed in the patients with DLP4 reveals individuals with the highest risk for atherosclerosis manifestation.
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http://dx.doi.org/10.1007/s11239-010-0474-4DOI Listing
January 2011

The role of thrombophilia in patients with retinal vein occlusion and no systemic risk factors.

Can J Ophthalmol 2010 Apr;45(2):171-5

Department of Ophthalmology, University of Leipzig, Germany.

Objective: The role of thrombophilia in the etiology of retinal vein occlusion (rVO) has not been adequately clarified. The aim of this study was to examine the prevalence of thrombophilia among RVO patients with and without systemic risk factors and among patients younger and older than 50 years.

Design: Prospective case-control study.

Participants: One hundred and twenty one patients with RVO, including 92 with acquired risk factors (hypertension, hyperlipidemia, and diabetes mellitus) and 29 without these factors. The control group included 60 persons matched for age, sex, and risk factors.

Methods: All participants were screened for Leiden mutation (FV Leiden), hyperprothrombinemia (20210 G/A mutation) and deficiency of protein C, S, and antithrombin.

Results: The prevalence of FV Leiden was significantly higher among RVO patients without risk factors (24.1%) than among controls without risk factors (0%; p = 0.034) and among RVO patients with acquired disorders (4.3%; p = 0.001). No significant differences were found in the prevalence of the other investigated thrombophilic factors. In all, 37.9% patients without acquired risk factors were positive for at least 1 thrombophilic factor compared with 7.6% RVO patients with acquired risk factors (p < 0.001) and 8.3% of the controls (p < 0.001). There was no significant difference in the prevalence of thrombophilic disorders among RVO patients according to age.

Conclusions: FV Leiden is significantly more frequent among RVO patients without acquired risk factors. Thrombophilia plays a much more important role in the pathogenesis of RVO in patients without acquired risk factors. Screening for thrombophilia is thus indicated only for patients in whom these factors have been excluded.
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http://dx.doi.org/10.3129/i09-273DOI Listing
April 2010

Molecular pathophysiology of thrombotic states and their impact to laboratory diagnostics.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2009 Mar;153(1):19-25

Department of Hemato-oncology, University Hospital, Olomouc, Czech Republic.

Background: Molecular genetic methods were implemented in the detection of thrombophilic disorders in the 1990's with the discovery of coagulation inhibitors antithrombin III (AT III), protein C (PC) and S (PS). The discovery of the molecular cause of activated protein C (APC) resistance by Bertina in 1994 greatly expanded their utilization.

Methods And Results: Currently, a broad group of molecular genetic markers with a clearly demonstrated risk of thrombophilia are used--mutation of FV Leiden 506R/Q, mutation of prothrombin (F II) 20210G/A, mutation of methylenetetrahydrofolate reductase (MTHFR) 677C/T in homozygous form, mutation of plasminogen activator inhibitor (PAI-1) 4G/5G, mutations of single coagulation inhibitors as well as a number of polymorphisms with controversial thrombophilic risk such as F XIII Val34Leu, platelet glycoproteins, endothelial protein C receptor and thrombomodulin. Another area utilizing molecular genetic methods is research of the pathophysiology of individual coagulation processes. To date, the greatest advances in regard to APC resistance have been achieved here. Although the molecular cause of APC resistance was clearly demonstrated in the 1990's, its clinical variability has not yet been fully explained. The same is true for the second most widespread mutation, prothrombin gene mutation, where only the latest research has hinted at a possible mechanism of expression of the genetic changes in the actual coagulation process.

Conclusions: The future of molecular genetic methods is in achieving a complex understanding of the pathophysiology of thrombophilia and not only in its utilization as a method for detecting many polymorphisms with a very low risk of thrombosis.
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http://dx.doi.org/10.5507/bp.2009.003DOI Listing
March 2009

The prevalence of activated protein C (APC) resistance and factor V Leiden is significantly higher in patients with retinal vein occlusion without general risk factors. Case-control study and meta-analysis.

Thromb Haemost 2008 May;99(5):925-9

Department of Ophthalmology, University Hospital Leipzig, Liebigstrasse 10-14, 04103 Leipzig, Germany.

Several small case-control studies have investigated whether factor V Leiden (FVL) is a risk factor for retinal vein occlusion (RVO) and generated conflicting data. To clarify this question we performed a large two-centre case-control study and a meta-analysis of published studies. Two hundred seven consecutive patients with RVO and a control group of 150 subjects were screened between 1996 and 2006. A systematic meta-analysis was done combining our study with further 17 published European case-control studies. APC resistance was detected in 16 out of 207 (7.7%) patients and eight out of 150 (5.3%) controls. The odds ratio (OR) estimated was 1.49 with a (non-significant) 95% confidence interval (CI) of 0.62-3.57. The meta-analysis including 18 studies with a total of 1,748 patients and 2,716 controls showed a significantly higher prevalence of FVL in patients with RVO compared to healthy controls (combined OR 1.66; 95% CI 1.19-2.32). All single studies combined in the meta-analysis were too small to reliably detect the effect individually. This explains the seemingly contradictory data in the literature. In conclusion, the prevalence of APC resistance (and FVL) is increased in patients with RVO compared to controls, but the effect is only moderate. Therefore, there is no indication for general screening of factor V mutation in all patients with RVO. We recommend this test to be performed in patients older than 50 years with an additional history of thromboembolic event and in younger patients without general risk factors like hypertension.
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http://dx.doi.org/10.1160/TH07-11-0658DOI Listing
May 2008

Endothelial haemostatic markers in members of families with familial combined hyperlipidemia.

Thromb Res 2009 16;123(3):466-75. Epub 2008 Apr 16.

3rd Department of Internal Medicine, University Hospital, I. P. Pavlova 6, Olomouc 77520, Czech Republic.

Introduction: The aim of this study was to evaluate the plasma levels of endothelial haemostatic markers - von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1), tissue plasminogen activator (t-PA) and soluble thrombomodulin (sTM) - in asymptomatic, nonsmoking members of families with familial combined hyperlipidemia (FCH). We investigated the association between these factors and the intima-media thickness (IMT) of the common carotid artery, selected risk factors of atherosclerosis and markers of insulin resistance.

Methods: 82 members of 29 FCH families were divided into two groups: HL (probands and hyperlipidemic first-degree relatives, n=47) and NL (normolipidemic first-degree relatives, n=35). The control groups C-HL (n=20) and C-NL (n=20) consisted of sex- and age-matched healthy individuals. IMT was measured by ultrasound at a far wall of both common carotid arteries.

Results: Compared with healthy controls, hyperlipidemic subjects had significantly higher levels of vWF (146.4+/-73.2% versus 112.2+/-29.4%, p<0.05), of PAI-1 (102.4[83.0-117.0] ng/ml versus 63.5[31.8-87.3] ng/ml, p<0.01) and of t-PA (5.1[2.5-7.9] ng/ml versus 3.4[1.4-5.8] ng/ml, p<0.05). They had increased IMT, which correlated with vWF (r=0.29, p<0.05). Their normolipidemic relatives had significantly higher levels of vWF (137.2+/-42.8% versus 106.6+/-24.0%, p<0.01) and of PAI-1 (75.3[53.2-92.0] ng/ml versus 48.6[37.4-85.9] ng/ml, p<0.05). Levels of vWF, PAI-l and t-PA were independently associated with several markers of insulin resistance.

Conclusions: Asymptomatic members of FCH families have increased endothelial haemostatic factors- vWF, PAI-1, t-PA, which are associated with insulin resistance. VWF correlates with morphological vascular changes, detected by the increase of IMT, presented in only hyperlipidemic subjects.
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http://dx.doi.org/10.1016/j.thromres.2008.02.011DOI Listing
April 2009

Frequency of selected thrombophilias in women with placental abruption.

Aust N Z J Obstet Gynaecol 2007 Aug;47(4):297-301

Department of Obstetrics and Gynecology, Medical Faculty of Palacký University, Olomouc, Czech Republic.

Objective: There is a growing view that inherited or acquired thrombophilia may predispose a woman towards an adverse pregnancy outcome. The aim of this study was to investigate whether risk factors for placental abruption because of such thrombophilias (such as carriership of factor V Leiden (FVL), prothrombin G20210A gene mutation and homozygous MTHFR C677T) might be used as a predictor for placental abruption.

Methods: A retrospective case-control study conducted at the University Hospital, Palacky University, Olomouc, Czech Republic. One hundred and eighty women with placental abruption out of 20,175 deliveries (0.79%) were compared to 196 unselected gravidae. A detailed medical history was taken with special reference to factors related to hypercoagulation and blood was drawn for polymerase chain reaction analysis. The prevalence of FVL, prothrombin G20210A and MTHFR C677T was related to placental abruption.

Results: The heterozygous form of FVL was present in 20of 142 cases (14.1%) in the placental abruption group, compared to ten of 196 (5.1%) in the control group (odds ratio 3.0, 95% confidence interval 1.4-6.7).

Conclusions: We found that factor V Leiden is a significant risk factor for placental abruption.
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http://dx.doi.org/10.1111/j.1479-828X.2007.00741.xDOI Listing
August 2007

Selected pregnancy variables in women with placental abruption.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2006 Nov;150(2):271-3

Department of Obstetrics and Gynecology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.

Objective: The aim of this study was to investigate risk factors for placental abruption and to determine if anamnestic variables such as inherited thrombosis or recurrent fetal loss might be used as a predictor for placental abruption.

Methods: A retrospective case-control study at the University Hospital, Palacky University, Olomouc, Czech Republic. One hundred and eighty women with placental abruptio out of 20,175 deliveries (0.79 %) who were compared to 196 unselected pregnant women. A detailed anamnesis was taken.

Results: Compared to controls, women with placental abruptio had a 12-fold increased prevalence of prior recurrent fetal loss and a 6-fold increased prevalence of inherited thrombosis.

Conclusions: We found that recurrent fetal loss, and inherited thrombosis may be significant risk factors for placental abruptio.
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http://dx.doi.org/10.5507/bp.2006.040DOI Listing
November 2006

Elevated thrombopoietin levels and alterations in the sequence of its receptor, c-Mpl, in patients with Diamond-Blackfan anemia.

Haematologica 2004 Nov;89(11):1391-2

In the study of the possible thrombopoietin (TPO)-c-Mpl pathway involvement in the pathogenesis of Diamond-Blackfan anemia, repeatedly increased serum TPO levels were identified in 7/14 patients and changes in c-mpl sequence in 3/14 patients. While elevated TPO levels can represent a compensatory mechanism for impaired erythropoiesis, c-Mpl mutations could influence the disease severity.
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November 2004

Endothelial damage and activation of the hemostatic system during radiofrequency catheter isolation of pulmonary veins.

J Interv Card Electrophysiol 2004 Jun;10(3):271-9

I. Department of Medicine, University Hospital, Olomouc, Czech Republic.

Aims: To determine the systemic thrombogenic effect of radiofrequency catheter isolation of the pulmonary veins (PVI) in the treatment of atrial fibrillation.

Methods And Results: We studied endothelial damage marker (von Willebrand factor [vWf]), fibrinolysis markers (tissue plasminogen activator [t-PA], plasminogen activator inhibitor-1 [PAI-1]) and coagulation activation markers (D-dimer [DD]) in 30 patients (pts) undergoing PVI. Heparin was administered continuously after double transseptal puncture in all pts. Concentrations of vWf and t-PA were significantly increased after accomplishing PVI compared to the baseline values, and elevated levels persisted 24 hours later ( p < 0.01). PAI-1 levels decreased following PVI compared to the baseline levels ( p = 0.02). PAI-1 levels normalized 24 hours after the procedure. DD increased continuously during the procedure with the peak following PVI ( p < 0.01). Higher DD concentrations persisted 24 hours later ( p = 0.02). In a multivariate analysis, total procedure time correlated significantly with the peak vWf and DD concentrations, while total RF energy dose correlated only with peak vWf ( r = 0.82). Time to heparin administration correlated with DD levels prior to the first RF pulse ( r = 0.83, p < 0.01) as well as after PVI ( r = 0.75, p < 0.01). A group of patients heparinized within the first hour of the PVI procedure had normal preablation DD levels and significantly mitigated DD levels following PVI compared to the group of patients heparinized later ( p < 0.01).

Conclusions: Pulmonary vein ablations cause an increased systemic procoagulant state as reflected by fibrin turnover, fibrinolysis activation and endothelial perturbation. The activation of the coagulation cascade could be decreased by early heparin administration.
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http://dx.doi.org/10.1023/B:JICE.0000026924.96281.beDOI Listing
June 2004
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