Inflammatory and oncogenic signaling converge in disease evolution of BCR-ABL-negative myeloproliferative neoplasms, clonal hematopoietic stem cell disorders characterized by gain-of-function mutation in JAK2 kinase (JAK2V617F), with highest prevalence in patients with polycythemia vera (PV). Despite the high risk, DNA-damaging inflammatory microenvironment, PV progenitors tend to preserve their genomic stability over decades until their progression to post-PV myelofibrosis/acute myeloid leukemia. Using induced pluripotent stem cells-derived CD34+ progenitor-enriched cultures from JAK2V617F+ PV patient and from JAK2 wild-type healthy control, CRISPR-modified HEL cells and patients' bone marrow sections from different disease stages, we demonstrate that JAK2V617F induces an intrinsic IFNγ- and NF-κB-associated inflammatory program, while suppressing inflammation-evoked DNA damage both in vitro and in vivo. We show that cells with JAK2V617F tightly regulate levels of inflammatory cytokines-induced reactive oxygen species, do not fully activate the ATM/p53/p21waf1 checkpoint and p38/JNK MAPK stress pathway signaling when exposed to inflammatory cytokines, suppress DNA single-strand break repair genes' expression yet overexpress the dual-specificity phosphatase (DUSP) 1. RNAi-mediated knock-down and pharmacological inhibition of DUSP1, involved in p38/JNK deactivation, in HEL cells reveals growth addiction to DUSP1, consistent with enhanced DNA damage response and apoptosis in DUSP1-inhibited parental JAK2V617F+ cells, but not in CRISPR-modified JAK2 wild-type cells. Our results indicate that the JAK2V617F+ PV progenitors utilize DUSP1 activity as a protection mechanism against DNA damage accumulation, promoting their proliferation and survival in the inflammatory microenvironment, identifying DUSP1 as a potential therapeutic target in PV.

• MeSH
• cytokiny genetika   metabolismus   MeSH
• fosfatasa 1 s dvojí specificitou genetika   MeSH
• hematopoetické kmenové buňky patologie   MeSH
• indukované pluripotentní kmenové buňky patologie   MeSH
• Janus kinasa 2 genetika   MeSH
• lidé MeSH
• mutace MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí MeSH
• oxidační stres * MeSH
• polycythaemia vera genetika   MeSH
• poškození DNA * MeSH
• proliferace buněk * MeSH
• reprodukovatelnost výsledků MeSH
• transkripční faktor STAT1 metabolismus   MeSH
• zánět metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH
• DUSP1 protein, human MeSH Prohlížeč
• fosfatasa 1 s dvojí specificitou MeSH
• JAK2 protein, human MeSH Prohlížeč
• Janus kinasa 2 MeSH
• STAT1 protein, human MeSH Prohlížeč
• transkripční faktor STAT1 MeSH

The effect of direct oral anticoagulants (DOACs) on laboratory tests dependent on the production of their targets, factor IIa and factor Xa (FXa), is a well-known problem and can cause both false positive and negative results. Therefore, the correct interpretation of tests performed in patients receiving DOACs is necessary to avoid misclassification and subsequent clinical consequences. However, even with significant experience, there are situations where it is not possible to assess the influence of some methods. Particularly important is the situation in the diagnosis of lupus anticoagulants using the dilute Russell viper venom timetest, which is based on direct FXa activation. A very promising solution to this situation is offered by the DOAC laboratory balancing procedure DOAC-Stop. For evaluating the effectiveness of this procedure, 60 (20 apixaban, 20 dabigatran, and 20 rivaroxaban) patients treated with DOACs were enrolled. All patient samples were analyzed for the presence of individual DOAC types and subsequently subjected to the DOAC-Stop procedure.We evaluated its effectiveness by our own high-performance liquid chromatography-coupled tandem mass spectrometrymethod, which simultaneously sets all high-sensitivity DOACs. Unlike coagulation tests based on the determination of the residual effects of DOACs on target enzymes, which is complicated by extensive interindividual variation, this methodology is highly specific and sensitive.The DOAC-Stop procedure eliminated dabigatran from 99.5%, rivaroxaban from 97.9%, and apixaban from 97.1% of participants in our group. Residual amounts did not exceed 2.7 ng/mL for dabigatran, 10.9 ng/mL for rivaroxaban, or 13.03 ng/mL for apixaban, which are safe values that do not affect either screening or special coagulation tests.

• Klíčová slova
• DOAC-Stop, HPLC MS/MS, anticoagulants, apixaban, dabigatran, factor Xa inhibitors, lupus inhibitor, rivaroxaban,
• MeSH
• antithrombiny MeSH
• chromatografie kapalinová metody   MeSH
• dabigatran analýza   farmakologie   terapeutické užití   MeSH
• hemokoagulace účinky léků   MeSH
• inhibitory faktoru Xa analýza   farmakologie   terapeutické užití   MeSH
• lidé MeSH
• lupusový inhibiční faktor koagulace krev   MeSH
• metody MeSH
• pyrazoly analýza   farmakologie   terapeutické užití   MeSH
• pyridony analýza   farmakologie   terapeutické užití   MeSH
• rivaroxaban analýza   farmakologie   terapeutické užití   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antithrombiny MeSH
• apixaban MeSH Prohlížeč
• dabigatran MeSH
• inhibitory faktoru Xa MeSH
• lupusový inhibiční faktor koagulace MeSH
• pyrazoly MeSH
• pyridony MeSH
• rivaroxaban MeSH

OBJECTIVE: To assess and compare the frequency of selected gene mutations of thrombophilic markers (FV Leiden, FII prothrombin G20210A and MTHFR C677T) in patients with primary and secondary infertility. DESIGN: Retrospective study. SETTING: Institute of normal anatomy, Faculty of Medicine and Dentistry, Palacky University Olomouc. METHODS: The study included 92 patients with primary infertility and 89 patients with secondary infertility. Indications for examination of these mutations were following: a positive family or personal history, a positive obstetrical history or a repeated failure of assisted reproduction treatment. RESULTS: According to our anticipation, women with the secondary infertility were significantly older(p < 0.0005) than those with primary infertility. No mutations of genes of examined thrombophilic markers (FV, FII and MTHFR), either alone or in combination, were found in only 8.7 % patients with primary infertility and in 5.6 % patients with secondary infertility. Significantly higher frequency of factor Leiden(p < 0.02) was observed in women with secondary infertility. There were no significant differences in the frequency of detected mutations of the remaining factors. CONCLUSION: Based on our findings we suggest that the assessment of selected gene mutations of thrombophilic markers should be a part of the diagnostic algorithm in patients with positive history for thrombophilic disorders.

• MeSH
• dospělí MeSH
• faktor V genetika   MeSH
• genetické markery MeSH
• lidé MeSH
• methylentetrahydrofolátreduktasa (NADPH2) genetika   MeSH
• mutace * MeSH
• protrombin genetika   MeSH
• trombofilie komplikace   genetika   MeSH
• ženská infertilita komplikace   genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• factor V Leiden MeSH Prohlížeč
• faktor V MeSH
• genetické markery MeSH
• methylentetrahydrofolátreduktasa (NADPH2) MeSH
• protrombin MeSH

In the Czech Republic, anagrelid is used according to the recommendations of the Czech Working Group on Myeloproliferative Disorders for treatment ofthrombocythaemias associated with chronic myeloproliferative disorders--mainly essential thrombocythaemia and, regularly, reports are being presented from the Register of Patients Treated with Thromboreductin, most recently last year (Vnitr Lék 2009; 55: I-XII). The Register commenced in 2005 and from then it aims to determine detailed clinical and laboratory profiles of the patients. The structure of the Register has changed significantly in the course of its existence, reflecting the reports from each of the analyses conducted so far. Also, the data entry in the database improves every year and it reaches 97% on some of the items. The longest evaluation period in some of the patients is 108 months. By April 2010, the Register database contained data on 717 patients. Of these, 672 patients with the diagnosis of a Ph-negative chronic myeloproliferative disorder were evaluated. This year's analysis included the patients with essential thrombocythaemia, polycythaemia vera and primary myelofibrosis only. The analysis included 418 women and 254 men with median age of50 years. Unlike the first years, 2/3 of the current sample are non pretreated patients, meaning that the patients reach the specialized centres early in their treatment. Also, patients, and the older patients in particular, are more frequently treated with combined regimens including Thromboreductin. We increasingly observe hypertension as one of the monitored risk factors preceding the disease and laboratory parameters showJAK2 mutation in more than a half of patients while some form ofthrombotic diathesis is found in the anamnesis of 7-10% of patients. Some bleeding is observed in 1-5% of the registered patients. In comparison to the previous years, this is a decrease in the prevalence of clinical symptoms prior to the disease onset; this is very likely associated with an earlier patient diagnosis within the asymptomatic phase of the disease. Therapeutically, we achieve a fast treatment response but there still are 16.3% of sufficient afterone year of treatment. Thromboreductin dose is increasing but even in this group it does not exceeds the mean of 2.38 mg per 24 hours. Complications are observed in 6.2% of patients in the first year of therapy, and ofthese, thrombotic events in about 2.5% and (small) bleeding complications in 4% of patients. The data suggest that we still do not reach treatment response in a certain proportion of patients after a year of their therapy. Even though the care results from the analysed data improve every year, the Register helps to uncover some issues that still remain, such as treatment intensification and other treatment modifications.

• MeSH
• chinazoliny škodlivé účinky   terapeutické užití   MeSH
• dospělí MeSH
• esenciální trombocytemie krev   farmakoterapie   MeSH
• inhibitory agregace trombocytů škodlivé účinky   terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• počet trombocytů MeSH
• polycythaemia vera krev   farmakoterapie   MeSH
• primární myelofibróza krev   farmakoterapie   MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• anagrelide MeSH Prohlížeč
• chinazoliny MeSH
• inhibitory agregace trombocytů MeSH

The registry of patients treated with Thromboreductin (anagrelide) in the Czech Republic contains data concerning patients that have been treated using this drug since 2004. As of June 2009, the total number of patients was 549. The current analysis focused mainly on evaluation of anagrelide dosage needed to achieve a complete response in high-risk patients: reduction in platelet count to below 400 x 10(9)/l, which was also considered as reaching the therapeutic goal. The outcomes of the registry confirm that although anagrelide (Thromboreductin) is a very effective platelet-reducing agent, the administration of which is related to a low incidence of adverse effects and complications, the therapeutic goal is not achieved in all cases and or despite a quick treatment response, the therapeutic goal is achieved more slowly.

• MeSH
• chinazoliny terapeutické užití   MeSH
• dospělí MeSH
• esenciální trombocytemie farmakoterapie   MeSH
• inhibitory agregace trombocytů terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• myeloproliferativní poruchy komplikace   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• trombocytóza komplikace   farmakoterapie   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
• klinické zkoušky MeSH
• Názvy látek
• anagrelide MeSH Prohlížeč
• chinazoliny MeSH
• inhibitory agregace trombocytů MeSH

Molecular genetic methods passed into the field of investigation of thrombophilic states in 90th years of last century, along with the first discoveries of coagulation inhibitors (AT III, protein C and protein S). They have acquired a widespread use above all with the detection of the molecular basis of activated protein C (APC) resistance in 1994 by prof. Bertina. At the present time, a wide range of molecular genetic markers, linked with a clearly documented increased risk of thrombophilia are adapted. They include mutations of factor V Leiden 506R/Q, of protrombin 20210G/A, MTHFR 677C/T in homozygous form, mutation of PAI-1 4G/5G, mutations of different coagulation inhibitors and finally a range of polymorphisms with still not precisely defined increased risk for thrombophilia (F XIII Val34leu, platelets glycopeproteins, endothelial protein C receptor and trombomodulin). From the methodological viewpoint, all these techniques are based on the principle polymerase chain reaction (PCR). In the last period of time, however there was a rapid evolution, allowing a significant improvement in their laboriousness. Nowadays, splitting with the aid of restriction endonucleases, real time PCR or allel specific primers for PCR. The second, where molecular genetic methods are currently under use, is pathophysiological investigation of the single coagulation processes. Here, in a fact, most significant progress has been in the field of APC resistance made elucidation. Although still in the 90th years of the past century the genetical cause of these coagulation disturbance was unequivocally documented its clinically heterozygous appears not yet fully understood at the moment. Similarly, in prothrombin mutation, only the latest investigations have outlined the probable mechanism of expression. Concerning the future evolution of molecular genetic methods, there can be observed a clear cut tendency to better understanding the pathophysiologic cause of thrombophilia in comparison with the searching for new coagulation defects which consecutively bear lesser a relative risk of thrombosis.

• MeSH
• faktor V genetika   MeSH
• genetické techniky * MeSH
• hemokoagulace MeSH
• lidé MeSH
• trombofilie krev   diagnóza   genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• faktor V MeSH

The registry of patients treated with Thromboreductine (anagrelid) in the contributing centres in the Czech Republic has been updated with data on the patients receiving this medication since 2004. The original purpose of the registry was to record responses to Thromboreductine therapy and adverse drug reactions in patients with essential thrombocytopenia. However, data on additional Ph negative myeloproliferations, as well as data on cytoreductive therapies other than exclusively that using Thromboreductine has also been recorded in the course of its compilation, including data on combined regimes. At present, the database contains data on 421 patients, and valid conclusions can be drawn if the level of data filling is enhanced. Evaluation has been currently focused on the analysis of the risk of development of clinical symptoms of thrombosis and on the standards of treatment from the viewpoint of the achieved treatment response. Analyses of data from the registry corroborate the special importance of the proof of JAK2 mutation, and of the test for factor V Leiden mutation, and of protein of S for the assessment of the risk of thromboembolic complications. The output of the analysis confirms that anagrelid is a very efficient thromboreductive agent the administration of which is associated with a low incidence of non-serious adverse effects (10.9%). However, in spite of a fast response to therapy, the therapeutic goal consisting in the reduction of the platelet count below 400 (or below 600) x 10(9)/l, i.e. the complete (or partial) treatment response, is relatively slow to achieve. This is likely to be due to lack of radical corrections in the dosage of the drug for different reasons.

• MeSH
• chinazoliny škodlivé účinky   terapeutické užití   MeSH
• esenciální trombocytemie krev   komplikace   farmakoterapie   MeSH
• fibrinolytika škodlivé účinky   terapeutické užití   MeSH
• hodnocení rizik MeSH
• inhibitory agregace trombocytů škodlivé účinky   terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• myeloproliferativní poruchy krev   komplikace   farmakoterapie   MeSH
• počet trombocytů MeSH
• trombocytopenie komplikace   MeSH
• trombóza etiologie   prevence a kontrola   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• anagrelide MeSH Prohlížeč
• chinazoliny MeSH
• fibrinolytika MeSH
• inhibitory agregace trombocytů MeSH

Since 2005, registers of patients treated with Thromboreductin (anagrelid) kept by some centres in the Czech Republic have been supplied with data concerning patients whose treatment with this preparation started in 2004. The purpose of the register is to record responses to therapy by Thromboreductin and adverse events in patients with essential thrombocytemia and other myeloproliferations, and to subsequently analyse the data. Another objective is to detect predisposition to clinical symptomatology and disease complications. Apart from thrombocyte count, additional risk factors are monitored. The database currently contains data for 336 patients. Initial analyses of data from the register point to the fact that anagrelid is a highly effective thromboreductive agent the administration of which is associated with relatively low incidence of adverse events (11.8 %) of mild and usually transitory nature. The therapeutic objective is attained at a relatively slow rate (according to overall stratification under 400 or under 600 x 10(9)/l thrombocytes), which is probably due to insufficient dose adjustment.

• MeSH
• chinazoliny škodlivé účinky   terapeutické užití   MeSH
• dospělí MeSH
• fibrinolytika škodlivé účinky   terapeutické užití   MeSH
• inhibitory agregace trombocytů škodlivé účinky   terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• myeloproliferativní poruchy krev   farmakoterapie   MeSH
• počet trombocytů MeSH
• polycythaemia vera krev   farmakoterapie   MeSH
• trombocytóza krev   farmakoterapie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• anagrelide MeSH Prohlížeč
• chinazoliny MeSH
• fibrinolytika MeSH
• inhibitory agregace trombocytů MeSH

Anagrelide hydrochloride is an effective drug used in patients with ET and other myeloproliferative disorders with thrombocythemia to selectively decrease the number of thrombocytes. Indications for use of anagrelide were described in detail in Czech medical literature. Since 2005 data concerning treatment with anagrelide in some medical clinics have been collected in patient register showing course of treatment from 2004, when the medicament obtained marketing authorization from State Institute for Drug Control to be used in the treatment of thrombocythemia in myeloproliferative disorders. Aim of patient register is to monitor medical effect of anagrelide therapy and incidence of adverse effects in patients with ET and other myeloproliferative disorders and subsequent analysis of collected data. At the moment patient register contains data from 154 patients.

• MeSH
• chinazoliny škodlivé účinky   terapeutické užití   MeSH
• esenciální trombocytemie farmakoterapie   MeSH
• inhibitory agregace trombocytů škodlivé účinky   terapeutické užití   MeSH
• lidé MeSH
• myeloproliferativní poruchy komplikace   MeSH
• trombocytóza komplikace   farmakoterapie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• anagrelide MeSH Prohlížeč
• chinazoliny MeSH
• inhibitory agregace trombocytů MeSH

The authors evaluated retrospectively in a group of 69 adult patients with Hodgkin's lymphoma the relationship between the beta-2-microglobulin serum level, basic demographic parameters (age, sex) and factors characterizing the extent (stage III and IV, "bulk" or mediastinal mass, number of affected areas of lymph nodes) and activity of the tumour (presence of B-symptoms, red cell sedimentation rate, haemoglobin, albumin and lactate dehydrogenase level, number of leucocytes and lymphocytes). They analyzed also the possible prognostic impact of beta-2-microglobulin on the therapeutic response risk of relapse and patient's survival. Methods of univariant statistical analysis confirmed the correlation of beta-2-microglobulin level with all investigated metric parameters of patients (advanced age, number of affected nodes, red cell sedimentation rate and lactate dehydrogenase level, lower albumin, haemoglobin level, numbers of leucocytes and lymphocytes). In multivariant analysis however the only independent metric markers significantly associated with an elevated protein level were more advanced age of the patients (P = 0.0002) and a lower number of leucocytes (P = 0.05). The values of beta-2-microglobulin was not influenced by the extent of the tumour (stage III and IV, "bulk" or mediastinal mass, higher number of affected areas of lymph nodes). Significantly more frequently elevated protein values were recorded in patients with manifestations of B symptoms associated with the diagnosis (P = 0.0003). Multivariant analysis did not prove the importance of the serum level of beta-2-microglobulin as a prognostic factor in the sense of predicted remission, development of a relapse or death in conjunction with progression of Hodgkin's lymphoma.

• MeSH
• beta-2-mikroglobulin krev   MeSH
• dospělí MeSH
• Hodgkinova nemoc krev   patologie   terapie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lymfatické metastázy MeSH
• mladiství MeSH
• nádorové biomarkery krev   MeSH
• počet leukocytů MeSH
• prognóza MeSH
• retrospektivní studie MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• beta-2-mikroglobulin MeSH
• nádorové biomarkery MeSH