Androgen deprivation therapy has long been the first-line treatment for hormone-sensitive prostate cancer (HSPC). After progression to castration-resistant prostate cancer (CRPC), androgen receptor pathway inhibitors (ARPIs) are commonly used. Recently, combined therapy with androgen deprivation and an ARPI has been recommended for metastatic HSPC patients. Novel markers are urgently needed for monitoring this disease and for making therapeutic decisions. Plasma samples were collected from 140 patients with either metastatic HSPC (n = 72) or CRPC (n = 68) before the start of ARPI therapy. Digital PCR was used to assess AR gene amplification, while the expression levels of miR-375 were measured by quantitative PCR. Sixteen other clinical markers were also evaluated, including prostate-specific antigen (PSA), chromogranin A (CGA), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), C-reactive protein (CRP), lymphocyte-to-monocyte ratio, and platelet count. A multivariate analysis, adjusted for age and metastatic dissemination, identified miR-375 expression and lymphocyte-to-monocyte ratio to be the independent negative predictors of ARPI therapy failure in CRPC patients. Regarding the HSPC patients, this article reports the primary finding of the independent negative predictive value of platelet count, CRP, and CGA for the failure of combined androgen deprivation therapy and ARPI.

• MeSH
• androgenní receptory genetika   MeSH
• C-reaktivní protein * metabolismus   MeSH
• chromogranin A * krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• metastázy nádorů MeSH
• mikro RNA * genetika   krev   MeSH
• nádorové biomarkery krev   MeSH
• nádory prostaty rezistentní na kastraci * krev   genetika   patologie   farmakoterapie   diagnóza   MeSH
• neúspěšná terapie MeSH
• prognóza MeSH
• prostatický specifický antigen * krev   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• trombocyty * metabolismus   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• androgenní receptory MeSH
• C-reaktivní protein * MeSH
• chromogranin A * MeSH
• mikro RNA * MeSH
• MIRN375 microRNA, human MeSH Prohlížeč
• nádorové biomarkery MeSH
• prostatický specifický antigen * MeSH

Whether the lung is a primary site of platelet (PLT) production is still disputed. To address this question, PLT parameters in blood before and after pulmonary circulation in humans, rats, and rabbits were assessed by automatic hematology analyzers; bone marrow and pulmonary megakaryocytes in humans, mice, rats, and rabbits were evaluated by immunohistochemical staining; and pulmonary megakaryocytes in humans were analyzed by single-cell RNA sequencing. We found that the mean number of PLTs in rats was nearly threefold greater than that in rabbits and humans. The PLT distribution width after pulmonary circulation in humans, rats, and rabbits was consistently less than that before pulmonary circulation. However, except for the PLT population in the left atrium of rats was significantly greater than that in the right ventricle (n=20), the PLT populations between the left and right atria of rats (n=19), rabbits (n=19), and humans (n=24), between the left atrium and right ventricle of rabbits (n=19), and between the inferior vena cava and radial artery of humans (n=93) had no differences. Moreover, megakaryocytes in the lungs of mice, rats, rabbits, and humans were mononuclear, were mainly located perivascularly, and accounted for approximately 3-5 ‰. Their numbers were significantly lower, and their sizes were smaller than those of bone marrow. Conclusively, the lung can produce PLTs, but it is not a primary site of PLT biogenesis. The capability of pulmonary PLT generation differs among species; at least in rats, it is greater than that in rabbits and humans.

• MeSH
• dospělí MeSH
• králíci MeSH
• krysa rodu Rattus MeSH
• lidé středního věku MeSH
• lidé MeSH
• megakaryocyty metabolismus   cytologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• plíce * cytologie   fyziologie   metabolismus   MeSH
• plicní oběh fyziologie   MeSH
• potkani Sprague-Dawley MeSH
• trombocyty * cytologie   fyziologie   metabolismus   MeSH
• trombopoéza * fyziologie   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• králíci MeSH
• krysa rodu Rattus MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

P2Y12 receptors on the platelet plasma membrane are targeted by several antiplatelets drugs. Although oligomerization and functioning of P2Y12 receptors depend on the membrane environment, little is known about their preferred membrane localization and the role of surrounding lipid composition, especially the arachidonic acids (ARA), which are abundant in platelets. Coarse-grained molecular dynamics simulations of platelet plasma membrane based on the lipidomics data were used to investigate the P2Y12 lipid environment and the involvement of ARA in its oligomerization in platelet plasma membranes. The platelet plasma membrane contains two types of lipids nanodomains: ordered, enriched in SM and cholesterol, and disordered, enriched in ARA-containing lipids. P2Y12 receptors prefer to localize in these ARA-rich domains and induce the sorting of the ARA-containing lipids in their vicinity. This ARA-rich environment promotes the oligomerization of P2Y12 receptors and stabilizes the protein-protein interfaces of oligomers. As summary, oligomerization of P2Y12 receptors is promoted in ARA-rich nano-domains of the platelet plasma membrane.

• Klíčová slova
• Arachidonic acid, Blood platelets, Molecular dynamics simulation, Protein Multimerization, Protein–lipid interaction, Purinergic receptor P2Y12,
• MeSH
• buněčná membrána * metabolismus   chemie   MeSH
• cholesterol metabolismus   chemie   MeSH
• kyselina arachidonová * metabolismus   chemie   MeSH
• lidé MeSH
• multimerizace proteinu MeSH
• purinergní receptory P2Y12 * metabolismus   chemie   MeSH
• simulace molekulární dynamiky * MeSH
• trombocyty * metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cholesterol MeSH
• kyselina arachidonová * MeSH
• P2RY12 protein, human MeSH Prohlížeč
• purinergní receptory P2Y12 * MeSH

Chelation is the rational treatment modality in metal overload conditions, but chelators are often non-selective and can, hence, cause an imbalance in the homeostasis of physiological metals including calcium and magnesium. The aim of this study was to develop an affordable, rapid but sensitive and precise method for determining the degree of chelation of calcium and magnesium ions and to employ this method for comparison on a panel of known metal chelators. Spectrophotometric method using o-cresolphthalein complexone (o-CC) was developed and its biological relevance was confirmed in human platelets by impedance aggregometry. The lowest detectable concentration of calcium and magnesium ions by o-CC was 2.5 μM and 2 μM, respectively. The indicator was stable for at least 110 days. Four and seven out of twenty-one chelators strongly chelated calcium and magnesium ions, respectively. Importantly, the chelation effect of clinically used chelators was not negligible. Structure-activity relationships for eight quinolin-8-ols showed improvements in chelation particularly in the cases of dihalogen substitution, and a negative linear relationship between pKa and magnesium chelation was observed. Calcium chelation led to inhibition of platelet aggregation in concentrations corresponding to the complex formation. A novel method for screening of efficacy and safety of calcium and magnesium ion chelation was developed and validated.

• Klíčová slova
• Chelator, Depletion, Methodology, Platelet, Selectivity,
• MeSH
• agregace trombocytů účinky léků   MeSH
• chelátory * chemie   MeSH
• hořčík * chemie   MeSH
• lidé MeSH
• preklinické hodnocení léčiv metody   MeSH
• trombocyty účinky léků   metabolismus   MeSH
• vápník * analýza   metabolismus   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• chelátory * MeSH
• hořčík * MeSH
• vápník * MeSH

BACKGROUND AND PURPOSE: Although the amphiphilic nature of the widely used antithrombotic drug Ticagrelor is well known, it was never considered as a membranotropic agent capable of interacting with the lipid bilayer in a receptor-independent way. In this study, we investigated the influence of Ticagrelor on plasma membrane lipid order in platelets and if this modulates the potency of Ticagrelor at the P2Y12 receptor. EXPERIMENTAL APPROACH: We combined fluorescent in situ, in vitro and in silico approaches to probe the interactions between the plasma membrane of platelets and Ticagrelor. The influence of Ticagrelor on the lipid order of the platelet plasma membrane and large unilamellar vesicles was studied using the advanced fluorescent probe NR12S. Furthermore, the properties of model lipid bilayers in the presence of Ticagrelor were characterized by molecular dynamics simulations. Finally, the influence of an increased lipid order on the dose-response of platelets to Ticagrelor was studied. KEY RESULTS: Ticagrelor incorporates spontaneously into lipid bilayers and affects the lipid order of the membranes of model vesicles and isolated platelets, in a nontrivial composition and concentration-dependent manner. We showed that higher plasma membrane lipid order in platelets leads to a lower IC50 value for Ticagrelor. It is shown that membrane incorporation of Ticagrelor increases its potency at the P2Y12 receptor, by increasing the order of the platelet plasma membrane. CONCLUSION AND IMPLICATIONS: A novel dual mechanism of Ticagrelor action is suggested that combines direct binding to P2Y12 receptor with simultaneous modulation of receptor-lipid microenvironment.

• Klíčová slova
• P2Y12, antiplatelet drug, lipid order, plasma membrane, platelet,
• MeSH
• antagonisté purinergních receptorů P2Y farmakologie   chemie   MeSH
• buněčná membrána * účinky léků   metabolismus   MeSH
• lidé MeSH
• lipidové dvojvrstvy metabolismus   MeSH
• membránové lipidy metabolismus   MeSH
• purinergní receptory P2Y12 * metabolismus   účinky léků   MeSH
• simulace molekulární dynamiky MeSH
• ticagrelor * farmakologie   chemie   MeSH
• trombocyty * účinky léků   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antagonisté purinergních receptorů P2Y MeSH
• lipidové dvojvrstvy MeSH
• membránové lipidy MeSH
• P2RY12 protein, human MeSH Prohlížeč
• purinergní receptory P2Y12 * MeSH
• ticagrelor * MeSH

BACKGROUND AND AIMS: It is well known that elevated cholesterol is associated with enhanced platelet aggregation and patients suffering from familial hypercholesterolemia (FH) have a high risk of thrombotic cardiovascular events. Although decreasing cholesterol level is associated with attenuation of platelet hyperactivity, there are currently no data on the effect of convertase subtilisin/kexin type 9 monoclonal antibodies (PCSK9ab) on platelet reactivity in FH. The aim of the study was to analyse the impact of different therapies including PCSK9ab on platelet aggregation in FH. METHODS: This study enrolled all 15 patients treated in the University Hospital Hradec Králové for FH. PCSK9ab have been administered in 12 of 15 patients while 8 patients were also undergoing lipid apheresis. Blood samples from all patients including pre- and post-apheresis period were tested for platelet aggregation triggered by 7 inducers, and the effect of 3 clinically used drugs (acetylsalicylic acid, ticagrelor and vorapaxar) was compared as well. RESULTS: Although apheresis decreased the reactivity of platelets in general, platelet responses were not different between non-apheresis patients treated with PCSK9ab and apheresis patients (post-apheresis values) with the exception of ristocetin. However, when compared to age-matched healthy population, FH patients had significantly lower platelet aggregation responses to 4 out of 7 used inducers and higher profit from 2 out of 3 used antiplatelet drugs even after exclusion of FH patients regularly receiving conventional antiplatelet treatment. CONCLUSION: This study showed for the first time the suitability of PCSK9ab treatment for reduction of platelet reactivity in FH patients.

• Klíčová slova
• ADP receptor, Acetylsalicylic acid, Antiplatelet, Dyslipidemia, Ticagrelor, Vorapaxar,
• MeSH
• agregace trombocytů * účinky léků   MeSH
• dospělí MeSH
• hyperlipoproteinemie typ II * krev   terapie   farmakoterapie   MeSH
• inhibitory agregace trombocytů * terapeutické užití   farmakologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• monoklonální protilátky terapeutické užití   farmakologie   MeSH
• PCSK9 inhibitory * MeSH
• proproteinkonvertasa subtilisin/kexin typu 9 * imunologie   MeSH
• senioři MeSH
• separace krevních složek * MeSH
• trombocyty účinky léků   metabolismus   imunologie   MeSH
• výsledek terapie 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
• inhibitory agregace trombocytů * MeSH
• monoklonální protilátky MeSH
• PCSK9 inhibitory * MeSH
• PCSK9 protein, human MeSH Prohlížeč
• proproteinkonvertasa subtilisin/kexin typu 9 * MeSH

AIM: The effect of platelet-rich autoplasma on endometrial thickness and receptor sensitivity to estrogen and progesterone. MATERIALS AND METHODS: This prospective clinical study included 200 patients. The participants in the study were divided into two groups. The first control group received hormone replacement therapy (HRT). The second study group received an intrauterine infusion of platelet-rich autoplasma (PRP group). On the 19th day of the menstrual cycle, an ultrasound examination was performed to assess endometrial thickness, as well as an immunohistochemical analysis to determine receptor sensitivity to estrogen and progesterone. RESULTS: In the course of the study, we found that the use of platelet-rich autoplasma increased the thickness of the endometrium by 0.85 mm; the average thickness of the endometrium in the group who received PRP therapy was 8.25 (8.25-8.61) mm; and in the group of patients who only received HRT, it was 7.40 (7.34-7.65) mm. The sensitivity of receptors to estrogen in the experimental group increased by 3.5, in the experimental group it was 75.00 (71.43-74.22), and in the control group it was 71.50 (67.05-70.85). The sensitivity of receptors to progesterone also increased by 9.0, in the experimental group it was 95.0 (91.4-93.8), and in the control group it was 86.0 (83.47-86.27). CONCLUSION: Due to the action of platelet factors, PRP therapy has a positive effect on the endometrium, increasing its thickness and improving its receptivity. Therefore, it can be concluded that this method can find great practical application to improve the outcomes of assisted reproductive technology programs.

• Klíčová slova
• infertility, platelet-rich autoplasma, thin endometrium,
• MeSH
• dospělí MeSH
• endometrium * diagnostické zobrazování   metabolismus   účinky léků   MeSH
• estrogeny MeSH
• lidé MeSH
• plazma bohatá na destičky MeSH
• progesteron * MeSH
• prospektivní studie MeSH
• receptory pro estrogeny metabolismus   MeSH
• receptory progesteronu metabolismus   MeSH
• trombocyty metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• estrogeny MeSH
• progesteron * MeSH
• receptory pro estrogeny MeSH
• receptory progesteronu MeSH

Bernard-Soulier syndrome (BSS) is a rare inherited disorder characterized by unusually large platelets, low platelet count, and prolonged bleeding time. BSS is usually inherited in an autosomal recessive (AR) mode of inheritance due to a deficiency of the GPIb-IX-V complex also known as the von Willebrand factor (VWF) receptor. We investigated a family with macrothrombocytopenia, a mild bleeding tendency, slightly lowered platelet aggregation tests, and suspected autosomal dominant (AD) inheritance. We have detected a heterozygous GP1BA likely pathogenic variant, causing monoallelic BSS. A germline GP1BA gene variant (NM_000173:c.98G > A:p.C33Y), segregating with the macrothrombocytopenia, was detected by whole-exome sequencing. In silico analysis of the protein structure of the novel GPIbα variant revealed a potential structural defect, which could impact proper protein folding and subsequent binding to VWF. Flow cytometry, immunoblot, and electron microscopy demonstrated further differences between p.C33Y GP1BA carriers and healthy controls. Here, we provide a detailed insight into its clinical presentation and phenotype. Moreover, the here described case first presents an mBSS patient with two previous ischemic strokes.

• Klíčová slova
• Bernard-Soulier syndrome, GP1BA, autosomal dominant, macrothrombocytopenia, monoallelic,
• MeSH
• alely * MeSH
• Bernardův-Soulierův syndrom krev   diagnóza   genetika   MeSH
• fenotyp * MeSH
• genetická predispozice k nemoci * MeSH
• genetická variace * MeSH
• genetické asociační studie MeSH
• imunofenotypizace MeSH
• lidé MeSH
• mutační analýza DNA MeSH
• počet trombocytů MeSH
• rodokmen MeSH
• trombocytopenie krev   diagnóza   MeSH
• trombocytový glykoproteinový komplex Ib-IX genetika   metabolismus   MeSH
• trombocyty metabolismus   ultrastruktura   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• adhesion receptor MeSH Prohlížeč
• trombocytový glykoproteinový komplex Ib-IX MeSH

The objective of the present study was to evaluate platelet mitochondrial oxygen consumption using high-resolution respirometry (HRR) and metabolic flux analysis (MFA) and to verify the effect of advanced age on these parameters. HRR was used to analyze permeabilized and intact platelets, MFA to measure oxygen consumption rates (OCR), extracellular acidification rates (ECAR) and ATP production rate in intact fixed platelets. Two groups of healthy volunteers were included in the study: YOUNG (20-42 years, n=44) and older adults (OLD; 70-89 years; n=15). Compared to YOUNG donors, platelets from group OLD participants displayed significantly lower values of oxygen consumption in the Complex II-linked phosphorylating and uncoupled states and the Complex IV activity in HRR protocols for permeabilized cells and significantly lower resting and uncoupled respirations in intact cells when analyzed by both methods. In addition, mitochondrial ATP production rate was also significantly lower in platelets isolated from older adults. Variables measured by both methods from the same bloods correlated significantly, nevertheless those acquired by MFA were higher than those measured using HRR. In conclusion, the study verifies compromised mitochondrial respiration and oxidative ATP production in the platelets of aged persons and documents good compatibility of the two most widely used methods for determining the global performance of the electron-transporting system, i.e. HRR and MFA.

• MeSH
• adenosintrifosfát metabolismus   MeSH
• analýza metabolického toku metody   MeSH
• buněčné dýchání MeSH
• dospělí MeSH
• energetický metabolismus * MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• mitochondrie metabolismus   MeSH
• mladý dospělý MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• spotřeba kyslíku MeSH
• stárnutí krev   metabolismus   MeSH
• trombocyty metabolismus   MeSH
• věkové faktory MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• adenosintrifosfát MeSH

Pluripotent stem cells of the bone marrow are stimulated by different cytokines to proliferation and differentiation into various types of blood cells. These cytokines are mostly glycoproteins. Erythropoietin stimulates stem cells to the formation of erythrocytes while colony-stimulating factors cause the formation of different types of white blood cells. Stem cell factors play an important role in the maintenance and survival of blood cells of all types. Thrombopoietin stimulates stem cells to proliferation and formation of blood platelets. Granulocyte colony-stimulating factor is probably the most important drug in use. It stimulates stem cells to the formation of neutrophile granulocytes. It is often used in recombinant forms such as filgrastim in the treatment of neutropenia in cancer chemotherapy or AIDS. Its pegylated conjugates such as pegfilgrastim are also available. Its activity can be supported by plerixafor, a small molecule - bicyclam derivative acting as an indirect agonist of stem cells factor. It acts as an antagonist of CXCR4 receptor activation of which brakes hematopoiesis. The treatment of conditions accompanied by thrombocytopenia such as idiopathic thrombocytopenic purpura is currently not performed by thrombopoietin but synthetic agonists of its receptor are preferred. Romiplostim is a peptibody. It consists of a protein part interacting with the thrombopoietin receptor which is, however, different from thrombopoietin, and of Fc fragment of immunoglobulin G1. In contrast, small molecule thrombopoietin receptor agonists represented by eltrombopag can be given orally unlike all of the above.

• Klíčová slova
• Colony-stimulating factors, eltrombopag., pegylation, plerixafor, romiplostim, stem-cell factors, thrombopoietin,
• MeSH
• benzoáty chemie   farmakologie   MeSH
• buněčná diferenciace účinky léků   MeSH
• faktory stimulující kolonie farmakologie   MeSH
• hydraziny chemie   farmakologie   MeSH
• knihovny malých molekul chemie   farmakologie   MeSH
• leukocyty mononukleární cytologie   účinky léků   metabolismus   MeSH
• lidé MeSH
• pyrazoly chemie   farmakologie   MeSH
• receptory thrombopoetinu agonisté   metabolismus   MeSH
• růstový faktor kmenových buněk farmakologie   MeSH
• trombocyty cytologie   metabolismus   MeSH
• trombopoéza účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• benzoáty MeSH
• eltrombopag MeSH Prohlížeč
• faktory stimulující kolonie MeSH
• hydraziny MeSH
• knihovny malých molekul MeSH
• pyrazoly MeSH
• receptory thrombopoetinu MeSH
• růstový faktor kmenových buněk MeSH