• MeSH
• dospělí MeSH
• elektroencefalografie metody   MeSH
• funkční lateralita fyziologie   MeSH
• lidé MeSH
• mapování mozku MeSH
• prsty ruky fyziologie   MeSH
• svaly fyziologie   MeSH
• temenní lalok fyziologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• srovnávací studie MeSH

AIMS: A challenge of pulmonary vein isolation (PVI) in catheter ablation for paroxysmal atrial fibrillation (PAF) is electrical reconnection of the PV. EFFICAS I showed correlation between contact force (CF) parameters and PV durable isolation but no prospective evaluation was made. EFFICAS II was a multicentre study to prospectively assess the impact of CF guidance for an effective reduction of PVI gaps. METHODS AND RESULTS: Pulmonary vein isolation using a radiofrequency (RF) ablation catheter with an integrated force sensor (TactiCathTM) was performed in patients with PAF. Operators were provided EFFICAS I-based CF guidelines [target 20 g, range 10-30 g, minimum 400 g s force-time integral (FTI)]. Conduction gaps were assessed by remapping of PVs after 3 months, and gap rate was compared with EFFICAS I outcome. At follow up, 24 patients had 85% of PVs remaining isolated, compared with 72% in EFFICAS I (P = 0.037) in which CF guidelines were not used. The remaining 15% of gaps correlated to the number of catheter moves at creating the PVI line, quantified as Continuity Index. For PV lines with contiguous lesions and low catheter moves, durable isolation was 81% in EFFICAS I and 98% in EFFICAS II (P = 0.005). At index procedure, the number of lesions was reduced by 15% in EFFICAS II vs. EFFICAS I. CONCLUSION: The use of CF with the above guidelines and contiguous deployment of RF lesions in EFFICAS II study resulted in more durable PVI in catheter ablation of PAF.

• MeSH
• analýza selhání vybavení MeSH
• chirurgie s pomocí počítače přístrojové vybavení   metody   MeSH
• design vybavení MeSH
• dospělí MeSH
• fibrilace síní diagnóza   chirurgie   MeSH
• katetrizační ablace přístrojové vybavení   metody   MeSH
• lidé MeSH
• mapování potenciálů tělesného povrchu přístrojové vybavení   metody   MeSH
• mechanický stres MeSH
• měniče tlakové MeSH
• převodní systém srdeční chirurgie   MeSH
• venae pulmonales chirurgie   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky kontrolované MeSH
• multicentrická studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH

Atomic force microscopy (AFM) is not only a high-resolution imaging technique but also a sensitive tool able to study biomechanical properties of bio-samples (biomolecules, cells) in native conditions-i.e., in buffered solutions (culturing media) and stable temperature (mostly 37 °C). Micromechanical transducers (cantilevers) are often used to map surface stiffness distribution, adhesion forces, and viscoelastic parameters of living cells; however, they can also be used to monitor time course of cardiomyocytes contraction dynamics (e.g. beating rate, relaxation time), together with other biomechanical properties. Here we describe the construction of an AFM-based biosensor setup designed to study the biomechanical properties of cardiomyocyte clusters, through the use of standard uncoated silicon nitride cantilevers. Force-time curves (mechanocardiograms, MCG) are recorded continuously in real time and in the presence of cardiomyocyte-contraction affecting drugs (e.g., isoproterenol, metoprolol) in the medium, under physiological conditions. The average value of contraction force and the beat rate, as basic biomechanical parameters, represent pharmacological indicators of different phenotype features. Robustness, low computational requirements, and optimal spatial sensitivity (detection limit 200 pN, respectively 20 nm displacement) are the main advantages of the presented method.

• MeSH
• biomechanika * MeSH
• biosenzitivní techniky MeSH
• kardiomyocyty cytologie   MeSH
• lidé MeSH
• mikroskopie atomárních sil * přístrojové vybavení   metody   MeSH
• pluripotentní kmenové buňky cytologie   MeSH
• preklinické hodnocení léčiv MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Form and function of the mitotic spindle depend on motor proteins that crosslink microtubules and move them relative to each other. Among these are kinesin-14s, such as Ncd, which interact with one microtubule via their non-processive motor domains and with another via their diffusive tail domains, the latter allowing the protein to slip along the microtubule surface. Little is known about the influence of the tail domains on the protein's performance. Here, we show that diffusive anchorage of Ncd's tail domains impacts velocity and force considerably. Tail domain slippage reduced velocities from 270 nm s-1 to 60 nm s-1 and forces from several piconewtons to the sub-piconewton range. These findings challenge the notion that kinesin-14 may act as an antagonizer of other crosslinking motors, such as kinesin-5, during mitosis. It rather suggests a role of kinesin-14 as a flexible element, pliantly sliding and crosslinking microtubules to facilitate remodeling of the mitotic spindle.

• MeSH
• aparát dělícího vřeténka metabolismus   MeSH
• kineziny izolace a purifikace   metabolismus   MeSH
• mikrotubuly metabolismus   MeSH
• mitóza fyziologie   MeSH
• optická pinzeta MeSH
• proteinové domény MeSH
• proteiny asociované s mikrotubuly genetika   izolace a purifikace   metabolismus   MeSH
• proteiny Drosophily izolace a purifikace   metabolismus   MeSH
• rekombinantní proteiny genetika   izolace a purifikace   metabolismus   MeSH
• Saccharomyces cerevisiae - proteiny genetika   izolace a purifikace   metabolismus   MeSH
• vazba proteinů fyziologie   MeSH
• zelené fluorescenční proteiny genetika   izolace a purifikace   metabolismus   MeSH
• Publikační typ
• audiovizuální média MeSH
• časopisecké články MeSH
• práce podpořená grantem MeSH

Specific allergen immunotherapy is frequently associated with adverse reactions. Several strategies are being developed to reduce the allergenicity while maintaining the therapeutic benefits. Peptide immunotherapy is one such approach. Methods for the simple and rapid identification of immunogenic epitopes of allergens (i.e. allergenic epitopes) are ongoing and could potentially lead to peptide-based vaccines. An epitope extraction technique, based on biofunctionalized magnetic microspheres self-organized under a magnetic field in a channel of a simple microfluidic device fabricated from polydimethylsiloxane, was applied in the isolation and identification of prospective allergenic epitopes. Similarly to chromatographic column separations, the easily replaceable plug of self-organized beads in the channel benefits especially from an even larger surface-to-volume ratio and an enhanced interaction of the surfaces with passing samples. Ovalbumin, the major protein of egg white and a typical representative of food allergens, was selected as the model molecule. Highly resistant ovalbumin was at first efficiently digested by a magnetic proteolytic reactor with trypsin treated with l-1-tosylamido-2-phenylethyl chloromethyl ketone and the second step, i.e. capture of allergenic epitopes from the mixture of peptides, was performed by a magnetic immunoaffinity carrier with orientedly immobilized rabbit anti-ovalbumin IgG molecules. Captured peptides were released with 0.05% trifluoroacetic acid. The elution fractions were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The peptide fragment of ovalbumin HIATNAVLFFGR (m/z: 1345.75, position: 371-382) was identified as a relevant allergenic epitope in this way. Such a microfluidic magnetic force-based epitope extraction technique applied in the epitope mapping of ovalbumin has the potential to be a significant step towards developing safe and cost-effective epitope-based vaccines.

• MeSH
• alergeny chemie   imunologie   MeSH
• epitopy analýza   MeSH
• financování organizované MeSH
• hmotnostní spektrometrie MeSH
• imunomagnetická separace metody   MeSH
• mapování epitopu metody   MeSH
• mikrofluidní analytické techniky metody   MeSH
• mikrosféry MeSH
• ovalbumin chemie   imunologie   MeSH
• potravinová alergie MeSH
• vakcíny MeSH

OBJECTIVES: Social policies help people to overcome various unfavourable living situations, such as unemployment, which may lead to health inequalities. The aim of this study is to examine how adults and children perceive the impact of social policies connected to unemployment on well-being in the household, and whether their views differ. METHODS: We obtained data from 123 stakeholders in Slovakia, 96 adults and 27 children aged 11-15 years. We used concept mapping, based on qualitative data collection and quantitative data analysis. RESULTS: We obtained four clusters related to: children and education; current workforce; disadvantaged groups; labour office support. Adults rated the current workforce as the most important and urgent, and children the disadvantaged groups. Contrasts were largest on the disadvantaged groups and on combining family life and working abroad which children rated as very important and urgent but adults less so. CONCLUSIONS: Stakeholders had many perceptions, which may help to improve social policies. Adults were more concerned about work, and children were more so about inequalities. In general, adults were more practical and individualistic, and children, more emphatic and idealistic.

• MeSH
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• nezaměstnanost psychologie   statistika a číselné údaje   MeSH
• rodinné vztahy psychologie   MeSH
• socioekonomické faktory MeSH
• veřejná politika * MeSH
• zaměstnanost zákonodárství a právo   psychologie   statistika a číselné údaje   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Slovenská republika MeSH

Current models of gene expression, which are based on single-molecule localization microscopy, acknowledge protein clustering and the formation of transcriptional condensates as a driving force of gene expression. However, these models largely omit the role of nuclear lipids and amongst them nuclear phosphatidylinositol phosphates (PIPs) in particular. Moreover, the precise distribution of nuclear PIPs in the functional sub-nuclear domains remains elusive. The direct stochastic optical reconstruction microscopy (dSTORM) provides an unprecedented resolution in biological imaging. Therefore, its use for imaging in the densely crowded cell nucleus is desired but also challenging. Here we present a dual-color dSTORM imaging and image analysis of nuclear PI(4,5)P2, PI(3,4)P2 and PI(4)P distribution while preserving the context of nuclear architecture. In the nucleoplasm, PI(4,5)P2 and PI(3,4)P2 co-pattern in close proximity with the subset of RNA polymerase II foci. PI(4,5)P2 is surrounded by fibrillarin in the nucleoli and all three PIPs are dispersed within the matrix formed by the nuclear speckle protein SON. PI(4,5)P2 is the most abundant nuclear PIP, while PI(4)P is a precursor for the biosynthesis of PI(4,5)P2 and PI(3,4)P2. Therefore, our data are relevant for the understanding the roles of nuclear PIPs and provide further evidence for the model in which nuclear PIPs represent a localization signal for the formation of lipo-ribonucleoprotein hubs in the nucleus. The discussed experimental pipeline is applicable for further functional studies on the role of other nuclear PIPs in the regulation of gene expression and beyond.

• MeSH
• buněčné jadérko metabolismus   MeSH
• DNA vazebné proteiny metabolismus   MeSH
• fosfatidylinositolfosfáty metabolismus   MeSH
• lidé MeSH
• mikroskopie MeSH
• nádorové buněčné linie MeSH
• RNA-polymerasa II metabolismus   MeSH
• vedlejší histokompatibilní antigeny metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In this work we evaluate the applicability of different atomic force microscopy (AFM) modes, such as Phase Shift Imaging, Atomic Force Acoustic Microscopy (AFAM) and Force Spectroscopy, for mapping of the distribution pattern of low-molecular-weight biomimetic groups on polymer biomaterial surfaces. Patterns with either random or clustered spatial distribution of bioactive peptide group derived from fibronectin were prepared by surface deposition of functional block copolymer nano-colloids and grafted with RGDS peptide containing the sequence of amino acids arginine-glycine-aspartic acid-serine (conventionally labeled as RGDS) and carrying biotin as a tag. The biotin-tagged peptides were labeled with 40nm streptavidin-modified Au nanospheres. The peptide molecules were localized through the detection of bound Au nanospheres by AFM, and thus, the surface distribution of peptides was revealed. AFM techniques capable of monitoring local mechanical properties of the surface were proved to be the most efficient for identification of Au nano-markers. The efficiency was successfully demonstrated on two different patterns, i.e. random and clustered distribution of RGDS peptides on structured surface of the polymer biomaterial.

• MeSH
• fibronektiny chemie   MeSH
• mechanické jevy * MeSH
• mikroskopie atomárních sil metody   MeSH
• nanokuličky ultrastruktura   MeSH
• povrchové vlastnosti MeSH
• streptavidin chemie   MeSH
• zlato MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• chirurgie s pomocí počítače metody   MeSH
• fibrilace síní diagnóza   chirurgie   MeSH
• katetrizační ablace metody   MeSH
• lidé MeSH
• mapování potenciálů tělesného povrchu metody   MeSH
• myokard MeSH
• srdeční komory diagnostické zobrazování   patofyziologie   chirurgie   MeSH
• zobrazování trojrozměrné * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH

Teleost fishes exhibit a breath-taking diversity of sex determination and differentiation mechanisms. They encompass at least nine sex chromosome systems with often low degree of differentiation, high rate of inter- and intra-specific variability, and frequent turnovers. Nevertheless, several mainly female heterogametic systems at an advanced stage of genetic differentiation and high evolutionary stability have been also found across teleosts, especially among Neotropical characiforms. In this study, we aim to characterize the ZZ/ZW sex chromosome system in representatives of the Triportheidae family (Triportheus auritus, Agoniates halecinus, and the basal-most species Lignobrycon myersi) and its sister clade Gasteropelecidae (Carnegiella strigata, Gasteropelecus levis, and Thoracocharax stellatus). We applied both conventional and molecular cytogenetic approaches including chromosomal mapping of 5S and 18S ribosomal DNA clusters, cross-species chromosome painting (Zoo-FISH) with sex chromosome-derived probes and comparative genomic hybridization (CGH). We identified the ZW sex chromosome system for the first time in A. halecinus and G. levis and also in C. strigata formerly reported to lack sex chromosomes. We also brought evidence for possible mechanisms underlying the sex chromosome differentiation, including inversions, repetitive DNA accumulation, and exchange of genetic material. Our Zoo-FISH experiments further strongly indicated that the ZW sex chromosomes of Triportheidae and Gasteropelecidae are homeologous, suggesting their origin before the split of these lineages (approx. 40-70 million years ago). Such extent of sex chromosome stability is almost exceptional in teleosts, and hence, these lineages afford a special opportunity to scrutinize unique evolutionary forces and pressures shaping sex chromosome evolution in fishes and vertebrates in general.

• MeSH
• Characiformes * genetika   MeSH
• lidé MeSH
• malování chromozomů MeSH
• mapování chromozomů MeSH
• molekulární evoluce MeSH
• pohlavní chromozomy genetika   MeSH
• srovnávací genomová hybridizace MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH