The goal-directed control of behaviour critically depends on emotional regulation and constitutes the basis of mental well-being and social interactions. Within a socioemotional setting, it is necessary to prioritize effectively the relevant emotional information over interfering irrelevant emotional information to orchestrate cognitive resources and achieve appropriate behavior. Currently, it is elusive whether and how different socioemotional stimulus dimensions modulate cognitive control and conflict resolution. Theoretical considerations suggest that interference effects are less detrimental when conflicting emotional information is presented within a "positive socioemotional setting" compared with a "negative socioemotional setting." Using event-related potentials (ERPs) and source localization methods, we examined the basic system neurophysiological mechanisms and functional neuroanatomical structures associated with interactive effects of different interfering facial, socioemotional stimulus dimensions on conflict resolution. We account for interactive effects of different interfering socioemotional stimulus dimensions on conflict resolution, i.e., we show how the socioemotional valence modulates cognitive control (conflict processing). The data show that conflicts are stronger and more difficult to resolve in a negative emotional task-relevant setting than in a positive emotional task-relevant setting, where incongruent information barely induced conflicts. The degree of emotional conflict critically depends on the contextual emotional valence (positive or negative) in which this conflict occurs. The neurophysiological data show that these modulations were only reflected by late-stage conflict resolution processes associated with the middle (MFG) and superior frontal gyrus (SFG). Attentional selection processes and early-stage conflict monitoring do not seem to be modulated by interactive effects of different interfering socioemotional stimulus dimensions on conflict resolution.

• MeSH
• dospělí MeSH
• emoce fyziologie   MeSH
• evokované potenciály fyziologie   MeSH
• lidé MeSH
• mapování mozku * MeSH
• mladý dospělý MeSH
• pozornost fyziologie   MeSH
• vyjednávání psychologie   MeSH
• výraz obličeje MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Collisions between replication and transcription machineries represent a significant source of genomic instability. RECQ5 DNA helicase binds to RNA-polymerase (RNAP) II during transcription elongation and suppresses transcription-associated genomic instability. Here, we show that RECQ5 also associates with RNAPI and enforces the stability of ribosomal DNA arrays. We demonstrate that RECQ5 associates with transcription complexes in DNA replication foci and counteracts replication fork stalling in RNAPI- and RNAPII-transcribed genes, suggesting that RECQ5 exerts its genome-stabilizing effect by acting at sites of replication-transcription collisions. Moreover, RECQ5-deficient cells accumulate RAD18 foci and BRCA1-dependent RAD51 foci that are both formed at sites of interference between replication and transcription and likely represent unresolved replication intermediates. Finally, we provide evidence for a novel mechanism of resolution of replication-transcription collisions wherein the interaction between RECQ5 and proliferating cell nuclear antigen (PCNA) promotes RAD18-dependent PCNA ubiquitination and the helicase activity of RECQ5 promotes the processing of replication intermediates.

• MeSH
• biologické modely MeSH
• DNA řízené RNA-polymerasy metabolismus   MeSH
• DNA vazebné proteiny metabolismus   MeSH
• DNA-dependentní DNA-polymerasy metabolismus   MeSH
• elongace genetické transkripce MeSH
• fyziologický stres genetika   MeSH
• genetická transkripce * MeSH
• HEK293 buňky MeSH
• helikasy RecQ metabolismus   MeSH
• interakční proteinové domény a motivy MeSH
• lidé MeSH
• multienzymové komplexy metabolismus   MeSH
• otevřené čtecí rámce genetika   MeSH
• prekurzory RNA genetika   MeSH
• proliferační antigen buněčného jádra metabolismus   MeSH
• protein BRCA1 metabolismus   MeSH
• rekombinasa Rad51 metabolismus   MeSH
• replikace DNA * MeSH
• ribozomální DNA metabolismus   MeSH
• ubikvitinace MeSH
• ubikvitinligasy metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Recent state-of-the-art methods developed for the analysis of polar xenobiotics from different types of biological matrices usually employ liquid chromatography with mass spectrometry. However, there are limitations when a small amount of sample mass is available. For example, individual benthic invertebrates or fish tissue samples often weigh less than 100 mg (e.g., brain, liver) but are necessary to understand environmental fate and bioaccumulation dynamics. We developed ultra-fast methods based on a direct sample introduction technique. This included coupling laser diode thermal desorption with atmospheric pressure chemical ionization mass spectrometry (LDTD-APCI-MS). We then quantitated a common selective serotonin reuptake inhibitor (citalopram) in brain tissues of individual juvenile fish after in vivo exposure to environmentally relevant concentration. Two mass spectrometric methods based on low (LDTD-APCI-triple quadrupole (QqQ)-MS/MS) and high (LDTD-APCI-high-resolution product scan (HRPS)) resolutions were developed and evaluated. Individual instrument conditions were optimized to achieve an accurate and robust analytical method with minimum sample preparation requirements. We achieved very good recovery (97-108%) across the range of 1-100 ng g-1 for LDTD-APCI-HRPS. LDTD-APCI-QqQ-MS/MS showed poorer performance due to interferences from the matrix at the lowest concentration level. LDTD-APCI ionization was successfully validated for analysis of non-filtered sample extracts. Evaluation of final methods was performed for a set of real fish brain samples, including comparison of LDTD-APCI-HRPS with a previously validated LC-heated electrospray ionization-HRPS method. This new LDTD-APCI-HRPS method avoids the chromatographic step and provides important benefits such as analysis of limited sample masses, lower total sample volume (typically μL), and reduction in analysis time per sample run to a few seconds. Graphical abstract.

• MeSH
• antidepresiva druhé generace analýza   MeSH
• chemické látky znečišťující vodu analýza   MeSH
• citalopram analýza   MeSH
• lasery polovodičové MeSH
• mozek - chemie * MeSH
• Oncorhynchus mykiss * metabolismus   MeSH
• potrava z moře (živočišná) analýza   MeSH
• selektivní inhibitory zpětného vychytávání serotoninu analýza   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

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.

• MeSH
• antikoagulancia * aplikace a dávkování   krev   izolace a purifikace   toxicita   MeSH
• aplikace orální MeSH
• chromatografie kapalinová metody   MeSH
• hmotnostní spektrometrie metody   MeSH
• lidé MeSH
• lineární modely MeSH
• reprodukovatelnost výsledků MeSH
• senzitivita a specificita MeSH
• vyšetření krevní srážlivosti metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

A total of 196 out of 209 polychlorobiphenyl (PCB) congeners were resolved using GC×GC-TOFMS with a non-polar/ionic liquid column series consisting of poly(50%-n-octyl-50%-methyl)siloxane and (1,12-di(tripropylphosphonium)dodecane bis(trifluoromethansulfonyl)amide) in the first and second dimension, respectively. It has been found that 13 PCB congeners overlap in five doublets (CB12+CB13, CB62+CB75, CB70+CB76, CB97+CB125 and CB153+CB168) and one triplet (CB90+CB101+CB113). All toxic, "dioxin like" congeners were separated with no interferences from any PCB congener. The 109 PCBs present in Aroclor 1242 and the 82 PCBs present in Aroclor 1260 were resolved GC×GC-TOFMS analysis on this column set.

• MeSH
• iontové kapaliny chemie   MeSH
• plynová chromatografie s hmotnostně spektrometrickou detekcí metody   MeSH
• polychlorované bifenyly chemie   izolace a purifikace   MeSH
• siloxany chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In the second part of this study, a systematic comparison was made between two ion fragmentation acquisition modes, namely data-independent acquisition (DIA) and DIA with ion mobility spectrometry (IMS) technology. These two approaches were applied to the analysis of 192 doping agents in urine. Group I included 102 compounds such as stimulants, diuretics, narcotics, and β2-agonists, while Group II contained 90 compounds included steroids, glucocorticoids, and hormone and metabolic modulators. Important method parameters were examined and compared, including the fragmentation, sensitivity, and assignment capability with the minimum occurrence of false positive hits. The results differed between Group I and II in number of detected fragments when exploring the MS/MS spectra. In Group I only 13%, while in the Group II 64% of the substances had a higher number of fragments in DIA-IMS mode vs. DIA. In terms of sensitivity, the performance of the two modes with and without activated IMS dimension was identical for about 50% of the doping agents. The sensitivity was higher without IMS, i.e. in simple DIA mode, for 20-40% of remaining doping agents. Despite this sensitivity reduction with IMS, 82% of compounds from both Groups met the minimum required performance level (MRPL) criteria of the World Anti-Doping Agency (WADA) when the DIA-IMS mode was applied. Automated data processing is important in routine doping analysis. Therefore, processing methods were optimized and evaluated for the prevalence of false peak assignments by analysing the target substances at different concentrations in urine samples. Overall, a significantly higher number of misidentified compounds was observed in Group II, with an almost 2-fold higher number of misidentifications in DIA compared to DIA-IMS. This result highlights the benefit of the IMS dimension to reduce the rate of false positive in screening analysis. The optimized UHPLC-IM-HRMS method was finally applied to the analysis of urine samples from administration studies including nine doping agents from both Groups. However, to limit the number of interferences from the biological matrix, an emphasis is needed on the adequate settings of the data processing method.

• MeSH
• doping ve sportu * MeSH
• glukokortikoidy MeSH
• iontová mobilní spektrometrie * MeSH
• narkotika MeSH
• steroidy MeSH
• tandemová hmotnostní spektrometrie MeSH
• Publikační typ
• časopisecké články MeSH

A new ultra high performance liquid chromatography with electrospray ionization time of flight mass spectrometry method for the selective and sensitive separation, identification, and determination of selected designer benzodiazepines (namely, pyrazolam, phenazepam, etizolam, flubromazepam, diclazepam, deschloroetizolam, bentazepam, nimetazepam, and flubromazolam) in human serum was developed. The separation of the studied designer benzodiazepines was achieved on C18 chromatographic column using gradient elution within 6 min without any significant matrix interferences. Liquid-liquid extraction with butyl acetate was applied for serum samples cleanup and preconcentration of studied designer benzodiazepines. The method was validated in terms of linearity, limit of detection, limit of quantification, matrix effects, specificity, precision, accuracy, recovery, and sample stability. The limit of detection values were 0.10-0.15 ng/mL. The method was applied to a spiked serum sample to demonstrate its applicability for systematic toxicology analysis. Furthermore, a capillary chromatographic method with micellar electrokinetic chromatography was used for the estimation of partition coefficients of studied designer benzodiazepines as important parameters to evaluate their pharmacological and toxicological properties.

• MeSH
• benzodiazepiny krev   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací * MeSH
• lidé MeSH
• limita detekce MeSH
• micely MeSH
• reprodukovatelnost výsledků MeSH
• vysokoúčinná kapalinová chromatografie * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Spatial light modulators have become an essential tool for advanced microscopy, enabling breakthroughs in 3D, phase, and super-resolution imaging. However, continuous spatial-light modulation that is capable of capturing sub-millisecond microscopic motion without diffraction artifacts and polarization dependence is challenging. Here we present a photothermal spatial light modulator (PT-SLM) enabling fast phase imaging for nanoscopic 3D reconstruction. The PT-SLM can generate a step-like wavefront change, free of diffraction artifacts, with a high transmittance and a modulation efficiency independent of light polarization. We achieve a phase-shift > π and a response time as short as 70 µs with a theoretical limit in the sub microsecond range. We used the PT-SLM to perform quantitative phase imaging of sub-diffractional species to decipher the 3D nanoscopic displacement of microtubules and study the trajectory of a diffusive microtubule-associated protein, providing insights into the mechanism of protein navigation through a complex microtubule network.

• MeSH
• časové faktory MeSH
• interferenční mikroskopie metody   statistika a číselné údaje   MeSH
• kovové nanočástice ultrastruktura   MeSH
• lidé MeSH
• mikroskopie atomárních sil MeSH
• mikroskopie fázově kontrastní metody   statistika a číselné údaje   MeSH
• mikrotubuly metabolismus   ultrastruktura   MeSH
• nanotechnologie MeSH
• nanotrubičky ultrastruktura   MeSH
• optické jevy MeSH
• počítačová simulace MeSH
• proteiny asociované s mikrotubuly metabolismus   MeSH
• proteiny buněčného cyklu metabolismus   MeSH
• Schizosaccharomyces pombe - proteiny metabolismus   MeSH
• světlo MeSH
• tubulin metabolismus   MeSH
• zlato MeSH
• zobrazování trojrozměrné metody   statistika a číselné údaje   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Linker for activation in T cells (LAT) is a critical regulator of T-cell development and function. It organises signalling events at the plasma membrane. However, the mechanism, which controls LAT localisation at the plasma membrane, is not fully understood. Here, we studied the impact of helix-breaking amino acids, two prolines and one glycine, in the transmembrane segment on localisation and function of LAT. Using in silico analysis, confocal and super-resolution imaging and flow cytometry, we demonstrate that central proline residue destabilises transmembrane helix by inducing a kink. The helical structure and dynamics are further regulated by glycine and another proline residue in the luminal part of LAT transmembrane domain. Replacement of these residues with aliphatic amino acids reduces LAT dependence on palmitoylation for sorting to the plasma membrane. However, surface expression of these mutants is not sufficient to recover function of nonpalmitoylated LAT in stimulated T cells. These data indicate that geometry and dynamics of LAT transmembrane segment regulate its localisation and function in immune cells.

• MeSH
• adaptorové proteiny signální transdukční chemie   genetika   metabolismus   MeSH
• buněčná membrána metabolismus   MeSH
• glycin genetika   metabolismus   MeSH
• interferenční mikroskopie MeSH
• Jurkat buňky MeSH
• konfokální mikroskopie MeSH
• lidé MeSH
• membránové proteiny chemie   genetika   metabolismus   MeSH
• mutace MeSH
• prolin genetika   metabolismus   MeSH
• proteinové domény MeSH
• sekundární struktura proteinů MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie aminokyselin MeSH
• simulace molekulární dynamiky MeSH
• T-lymfocyty metabolismus   MeSH
• vápník metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• peritonitida patologie   terapie   MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• patologie
• gastroenterologie