INTRODUCTION: Chronic thromboembolic pulmonary hypertension (CTEPH) and venous thromboembolism (VTE) are thought to share many common risk factors. Our study aimed to determine the frequencies of 5 thrombosis-related gene single nucleotide polymorphisms (SNPs) associated with VTE in patients with CTEPH (n 129) compared with a control group of healthy individuals without a history of VTE (n 2637). METHODS: The SNPs of the following genes were investigated: F5 (F V Leiden, rs6025), F2 prothrombin (rs1799963), fibrinogen gamma (FGG, rs2066865), F11 (rs2289252) and ABO (non-O, rs8176719) in both groups. RESULTS: The study found that the rs1799963 variant was more common in patients with chronic thromboembolic pulmonary hypertension (CTEPH) compared to the control group (p < .0001). The GA heterozygous variant showed a significant increase with an odds ratio (OR) of 4.480 (95% CI: 2.344-8.562) or a finding by maximum likelihood analysis (MLA) with p < .0001. Additionally, there was a notable increase in the rs8176719 variant with p < .0001 in CTEPH patients. Both the homozygous G/G variant and the heterozygous -/G variant also showed an increase, with OR of 4.2317 (95% CI: 2.45571-7.2919) and 2.4324 (95% CI: 1.46435-4.0403) respectively, or MLA (p < .0001 and p .0006). The study also revealed a higher prevalence of the heterozygous C/T variant of rs2289252 in CTEPH patients, with an OR of 1.5543 (95% CI: 1.02503-2.3568) or MLA (p .0379). CONCLUSION: The study suggests that the observed gene polymorphisms F2 (rs1799963), ABO (rs8176719), and F11 (rs2289252) may play a role as independent heritable risk factors in the development of CTEPH.

• MeSH
• ABO systém krevních skupin genetika   MeSH
• chronická nemoc MeSH
• dospělí MeSH
• faktor V genetika   MeSH
• fibrinogen genetika   MeSH
• incidence MeSH
• jednonukleotidový polymorfismus * MeSH
• lidé středního věku MeSH
• lidé MeSH
• plicní embolie genetika   MeSH
• plicní hypertenze * genetika   MeSH
• protrombin genetika   MeSH
• senioři MeSH
• trombofilie genetika   MeSH
• žilní tromboembolie genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

High incidence of thrombosis and venous thromboembolism was reported in patients with COVID-19. In this study, we focused on analysis of thrombophilic mutations performed without a standard DNA extraction step. In one hundred of COVID-19 positive outpatients, real-time PCR for Leiden mutation in the FV gene and G20210A mutation in the FII gene was carried out from DNA extracts and modified whole blood samples, and their cycle threshold (Ct) values were evaluated. In the extracts, healthy homozygotes (wt/wt), heterozygotes (M/wt), and homozygous carriers of Leiden mutation (M/M) provided median Ct values of 18.5, 19.4/22.0, and 20.9. In the whole blood, Ct values were 25.3 (wt/wt), 24.8/27.2 (M/wt), and 26.9 (M/M). Median Ct values for G20210A in the extracts were 19.6 for homozygotes (wt/wt), and 19.7/20.4 for heterozygous carriers. The whole blood samples provided Ct values of 23.9 in healthy homozygotes and 26.3/27.2 in heterozygotes for G20210A mutation. No homozygous subjects for G20210A and no double heterozygotes (for Leiden and G20210A mutations) were found. Despite significant differences in the Ct values, genotyping showed complete result concordance of the DNA extracts and the whole blood samples. The integrity and amplificability of DNA molecules in the whole blood samples during 28 days of deep freezing, interrupted by four cycles of thawing, did not significantly change. In conclusion, we demonstrated a new protocol for the detection of the thrombophilic mutations via real time PCR on the modified whole blood of COVID-19 positive patients. The blood modification was reliable, easy, cheap, and saving costs and turnaround time of the whole laboratory process.

• MeSH
• COVID-19 * diagnóza   genetika   MeSH
• DNA MeSH
• faktor V genetika   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• mutace MeSH
• protrombin genetika   MeSH
• rizikové faktory MeSH
• SARS-CoV-2 genetika   MeSH
• testování na COVID-19 MeSH
• trombofilie * genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

AIM: The aim of this study was to evaluate the clinical contribution of protein induced by vitamin K absence (PIVKA-II) for hepatocellular carcinoma (HCC) diagnostics and compare it with alpha-foetoprotein (AFP), a routinely used tumour marker. MATERIALS AND METHODS: A total of 332 participants were enrolled in this study: 64 with HCC, 48 with liver metastases of colorectal cancer origin, 42 with liver cirrhosis and 178 healthy individuals. Serum levels of PIVKA-II were measured using the chemiluminescent assay of the Architect 1000i System (Abbott, USA) and AFP levels using the chemiluminescent assay by DxI 800 (Beckman Coulter, USA). RESULTS: PIVKA-II achieved better clinical sensitivity than AFP and the difference in this sensitivity was statistically significant. PIVKA-II achieved the best sensitivity (96.9%) in distinguishing between the HCC and control groups with the proposed cut-off value of 60 mAU/ml. CONCLUSION: Our recommendation is for addition of PIVKA-II to the routine panel of HCC tumour markers.

• MeSH
• biologické markery krev   MeSH
• diferenciální diagnóza MeSH
• dospělí MeSH
• hepatocelulární karcinom krev   genetika   patologie   MeSH
• jaterní cirhóza krev   genetika   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové biomarkery krev   MeSH
• nádory jater krev   genetika   patologie   MeSH
• pilotní projekty MeSH
• proteinové prekurzory krev   genetika   MeSH
• protrombin genetika   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The incidence of acute myocardial infarction (AMI) increases with clustering of predisposing risk factors. In younger subjects with a positive family history of AMI occurring in relatives under the age of 60 years without obvious risk factors for atherosclerosis, there is a potential for strong inherited traits contributing to the risk of coronary disease. Among them there is increasing evidence that hereditary thrombophilia may play a major role. We present a unique case of a patient developing AMI at the age of 48 years. In this patient, without traditional risk factors for atherosclerosis, eight mutations and polymorphisms in six different genes were identified: polymorphism of factor V Leiden (1691 GA), factor II prothrombin (20210 GA), methylenetetrahydrofolate reductase (MTHFR, 677 CT and 1298 AC), plasminogen activator inhibitor 1 (PAI-1) polymorphism 4G/5G and glycoprotein VI (GP6, 13254 TC, Ser219Pro). All could be involved in the pathogenesis of the arterial thrombosis. Although such associations are extremely rare, it underlines the importance of thrombophilia assessment in cases with otherwise unexpected coronary disease occurring at young age. According to our experience, in the case of documented hereditary thrombophilia lineal relatives should be examined and/or followed up.

• MeSH
• antigeny genetika   MeSH
• ateroskleróza genetika   MeSH
• faktor V genetika   MeSH
• genetická predispozice k nemoci genetika   MeSH
• glykoproteiny membrány trombocytů genetika   MeSH
• infarkt myokardu genetika   MeSH
• inhibitor aktivátoru plazminogenu 1 genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• methylentetrahydrofolátreduktasa (NADPH2) genetika   MeSH
• mutační analýza DNA * MeSH
• polymorfismus genetický genetika   MeSH
• protrombin genetika   MeSH
• rizikové faktory MeSH
• trombofilie genetika   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH

Úvod: Mutácia faktora V Leiden (FVL) spolu a mutácia G20210A v protrombínovom géne (PTM) patria medzi 2 najčastejšie genetické polymorfizmy, ktoré predispozíciou pre rozvoj prevej epizódy venózneho tromboemblizmu (VTE). PTM sa vyskytuje v 2 % belošskej populácie. Hlavným cieľom tejto práce bolo zistiť prevalenciu PTM v populácii pacientov s anamnézou trombotickej príhody vs. zdravých kontrolách. Materiál a metódy: Za účelom posúdenie prítomnosti PTM bola realizovaná PCR analýza z DNA extrahovanej z periférnych leukocytov. Výsledky: Do štúdie bolo zaradených 2 274 pacientov, z nich 157 (6,9 %) malo prítomnú PTM. PTM mutácia bola prítomná u 2,6 % kontrol z celkového počtu 303 dobrovoľníkov. Analyzovali sme klinickú manifestáciu PTM. Pozorovali sme 123 venóznych trombóz, 46 artériových trombóz a 14 opakovaných spontánnych potratov. V tomto článku sme ďalej analyzovali ďalšie možné rizikové faktory rozvoja trombózy u pacientov s prítomnou PTM. Záver: Podľa našich vedomostí je toto najväčšia epidemiologická štúdia zameraná na výskyt PTM v strednej Európe. Za použitia štatistickej analýzy sme zistili relatívne vysoký výskyt PTM v populácii pacientov s anamnézou trombózy (6,9 %), ale aj u zdravých kontrol (2,6 %). Riziko trombózy je nezávislé od veku a pohlavia. Štúdia zároveň ukázala pomerne častý výskyt dvojitej prítomnosti PTM a FVL.

Introduction: Factor V Leiden (FVL) and prothrombin G20210A mutation (PTM) are the two most common genetic polymorphisms known to predispose to a first episode of venous thromboembolism (VTE). PTM is present in 2 % Caucasian population. The main aim of this study was to identify the PTM in the patients with positive history of thrombotic events vs. control subjects. Materials and Methods: The assessment of PTM was performed by the PCR analysis of the chromosomal DNA, which was isolated from the peripheral blood leukocytes. Results: Of the 2 274 patients included, 157 (6.9 %) were carriers of the PTM. The mutation was present only in 2.6 % (n = 8) of the 303 controls. The following clinical manifestations of PTM were analysed. We observed 123 venous thrombotic events, 46 arterial thrombosis and 14 spontaneous abortions. In this article we analyse other possible risk factors for thromboembolic events in patients with carriage of PTM. Conclusions: To our knowledge, this is the largest epidemiological study of PTM in Central Europe. Employing statistical analysis, we found relatively high prevalence of the PTM in both, the patients with positive thrombosis history (6.9 %), as well as in the control group (2.6 %). The risk of thrombosis by carriage of PTM is independent of age and gender. Study has shown relatively frequent presence of double carriership of PTM and factor V Leiden mutation (FVL).

• MeSH
• dospělí MeSH
• epidemiologické studie MeSH
• interpretace statistických dat MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mutace genetika   MeSH
• mutační analýza DNA MeSH
• polymerázová řetězová reakce MeSH
• polymorfismus genetický MeSH
• protrombin * genetika   MeSH
• statistika jako téma MeSH
• tromboembolie * genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Geografické názvy
• Slovenská republika MeSH

The aim of our study was to analyse inherited thrombotic disorders that influence the long-term outcome of PTA. Methods. Diabetic patients with peripheral arterial disease (PAD) treated by PTA in our centre between 2008 and 2011 were included in the study. Patients were divided into unsuccessful PTA group (75 patients), successful PTA group (58 patients), and control group (65 patients, with diabetes but no PAD). Diagnosis of inherited thrombotic disorders included mutation in factor V (Leiden), factor II (prothrombin), and mutation in genes for methylenetetrahydrofolate reductase-MTHFR (C677T and A1298C). Results. The genotypic frequency of Leiden allele G1691A was significantly associated with a risk of unsuccessful PTA in comparison with successful PTA group and control group (OR 8.8 (1.1-70.6), p = 0.041, and OR 9.8 (1.2-79.2), p = 0.032, resp.). However, we only observed a trend for the association of the prothrombin allele G20210A and risk of PTA failure. The frequencies of alleles of MTHFR 677 or 1298 did not differ significantly among the groups. Conclusion. Our study showed higher frequency of heterozygous form of Leiden mutation in diabetic patients with unsuccessful outcome of PTA in comparison with patients with successful PTA and diabetic patients without PAD.

• MeSH
• angioplastika škodlivé účinky   MeSH
• bérec krevní zásobení   MeSH
• bodová mutace * MeSH
• diabetické angiopatie etiologie   patofyziologie   terapie   MeSH
• faktor V analýza   genetika   MeSH
• genetické asociační studie MeSH
• ischemie epidemiologie   etiologie   prevence a kontrola   MeSH
• lidé středního věku MeSH
• lidé MeSH
• methylentetrahydrofolátreduktasa (NADPH2) krev   genetika   MeSH
• paže krevní zásobení   MeSH
• prevalence MeSH
• protrombin analýza   genetika   MeSH
• průřezové studie MeSH
• recidiva MeSH
• senioři MeSH
• substituce aminokyselin MeSH
• trombofilie krev   komplikace   genetika   patofyziologie   MeSH
• trombóza komplikace   etiologie   patofyziologie   terapie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky kontrolované MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

• MeSH
• diagnostické techniky molekulární * MeSH
• genetické testování * MeSH
• hodnocení rizik MeSH
• konsensus MeSH
• lidé MeSH
• mutace MeSH
• protrombin genetika   MeSH
• rezistence k aktivovanému proteinu C genetika   MeSH
• společnosti lékařské MeSH
• tromboembolie * komplikace   MeSH
• trombofilie * diagnóza   genetika   komplikace   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• zprávy MeSH

OBJECTIVE: Due to contradictory results of previous studies evaluating the association between ischemic stroke (IS) and thrombophilic polymorphisms, their routine screening in IS patients, particularly those older than 60 years, is not recommended. We evaluated the differences in the distribution of rs6025 and rs1799963 polymorphisms according to IS subtypes and their interaction with smoking. METHODS: We conducted a case-control study of 423 hospital-based consecutive survivors of their first-ever IS and 614 population-based controls. Survivors (18-81 years) with IS documented by brain imagining were examined at a median of 16 months after the index event. The stroke subtype was categorized using the Causative Classification of Stroke System. Controls (50-75 years) were free of a history of stroke/TIA, coronary heart disease, and venous thromboembolism. RESULTS: Age- and gender-adjusted prevalence of individuals carrying at least one copy of the rs1799963A minor allele was 5.3% among stroke survivors (by subtypes: 3.1% in large artery atherosclerosis, 2.0% in cardio-aortic embolism, 2.4% in small artery occlusion, and 10.3% in undetermined stroke) vs. 2.4% among controls. In multinomial multivariate adjusted analysis, rs1799963 was exclusively associated with undetermined stroke (OR: 3.67; 95% CI: 1.52-8.85; p = 0.004). There was strong evidence of rs1799963 × smoking synergistic interaction (OR: 5.14; 95% CI: 1.65-16.01; p = 0.005). There was no association of rs6025 with IS in general, or with any subtype. CONCLUSIONS: In our consecutive IS survivors, carriage of the rs1799963A allele is associated with undetermined stroke. This effect appears to be confined to smokers.

• MeSH
• cévní mozková příhoda diagnóza   epidemiologie   genetika   MeSH
• dospělí MeSH
• fenotyp MeSH
• frekvence genu MeSH
• genetická predispozice k nemoci MeSH
• heterozygot MeSH
• hodnocení rizik MeSH
• homozygot MeSH
• ischemie mozku diagnóza   epidemiologie   genetika   MeSH
• kouření škodlivé účinky   epidemiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• logistické modely MeSH
• mladiství MeSH
• mladý dospělý MeSH
• multivariační analýza MeSH
• odds ratio MeSH
• polymorfismus genetický * MeSH
• prevalence MeSH
• protrombin genetika   MeSH
• rizikové faktory MeSH
• rozdělení chí kvadrát MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• trombofilie krev   komplikace   diagnóza   epidemiologie   genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

OBJECTIVE: To evaluate the course of pregnancy and puerperium in asymptomatic carriers of FV Leiden and FII prothrombin mutation in heterozygous configuration in terms of risk of thrombembolic disease (TED) and late pregnancy complications. To evaluate whether global prophylactic LMWH administration during pregnancy benefits these women. METHODS: We monitored the incidence of thrombembolic events and severe late pregnancy complications in 473 asymptomatic carriers of FV Leiden and FII prothrombin mutation in heterozygous configuration. In 253 women, preventive LMWH application was introduced already during pregnancy. In 220 women, the application of LMWH was commenced as late as on the delivery day. In both groups application of LMWH continued during the puerperium. RESULTS: The incidence of TED in the whole group of carriers of thrombophylic mutations accounted for 0.19%. The incidence of severe late pregnancy complications was low - 2.5% compared with general population of pregnant women (6.4%). CONCLUSIONS: No direct causal relationship was established between asymptomatic carriage of Leiden and prothrombin mutation in heterozygous configuration and the occurrence of severe late pregnancy complications. There was no benefit from general LMWH prophylaxis started as early as pregnancy in these women and thus we consider it unnecessary.

• MeSH
• antikoagulancia terapeutické užití   MeSH
• faktor V genetika   MeSH
• heparin nízkomolekulární terapeutické užití   MeSH
• heterozygot * MeSH
• incidence MeSH
• kardiovaskulární komplikace v těhotenství epidemiologie   genetika   prevence a kontrola   MeSH
• komplikace porodu epidemiologie   genetika   prevence a kontrola   MeSH
• lidé MeSH
• mutace MeSH
• poporodní období MeSH
• protrombin genetika   MeSH
• rizikové faktory MeSH
• těhotenství MeSH
• tromboembolie epidemiologie   genetika   prevence a kontrola   MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

AIMS: TheAIM of our study is to present a novel approach for preparing a compound heterozygous reference material (hetRM) using gene synthesis technology with inverted insertion of wild-type and mutant fragments into a single cloning vector. Factor II (G20210A) and Factor V (G1691A Leiden) gene mutations were used as an experimental model. METHODS: During the gene synthesis, DNA fragments were aligned in the following order: G1691 FV wild-type forward strain, G20210 FII wild-type forward strain, 1691A FV mutant reverse strain, 20210A FII mutant reverse strain. The complete chain was inserted into a pIDT SMART cloning vector and amplified in an E. coli competent strain. For assessing hetRM characteristics and commutability, we used real-time PCR with subsequent melting curve analysis, real-time PCR with hydrolysis probes, allele-specific amplification, reverse hybridization, and dideoxynucleotide DNA sequencing. RESULT: All five methods yielded concordant results of DNA analysis of the hetRM. Differences in real-time PCR cycle threshold values after six-months of storage at -80 °C were not statistically significant from those obtained from freshly prepared hetRM aliquots, which is a good indication of their stability. CONCLUSION: By applying the procedures of gene synthesis and cloning technology, we prepared and verified a model genetic reference material for FII G20210A and FV G1691A testing with a compound heterozygous genotype. The hetRM was stable, commutable, and available in large quantities and in a wide concentration range.

• MeSH
• Escherichia coli MeSH
• faktor V genetika   MeSH
• genetické techniky * MeSH
• klonování organismů metody   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• mutace MeSH
• protrombin genetika   MeSH
• sekvenční analýza DNA MeSH
• trombofilie genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH