Current management of patients with AIS relies primarily on conservative approach. Suggested streamlined fast-track single-stop approach should expedite the workflow and improve clinical outcomes. Given the broadly available interventional cardiology expertise cardiologist should play an active and sensible role in the development of new interventional concepts of care of patients with acute ischaemic stroke.

• MeSH
• čas zasáhnout při rozvinutí nemoci MeSH
• cévní mozková příhoda terapie   MeSH
• embolektomie metody   MeSH
• infarkt myokardu terapie   MeSH
• lidé MeSH
• reperfuze myokardu metody   MeSH
• revaskularizace myokardu metody   MeSH
• trombolytická terapie metody   MeSH
• záchranná terapie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Chromosome architecture needs to be investigated in relation with the chemical function of DNA. The kinetics of gene expression, DNA replication, and repair are driven by the mechanisms by which a functional nuclear protein finds its substrate in the nucleus. Single-particle tracking (SPT) is a method to quantify fluorescent molecules dynamics from the tracks of the single molecules recorded by high-resolution microscopes. SPT offers direct observation of the movement and single-molecule resolution. Usually SPT is performed on membranes because of higher contrast. Here, we introduce a novel method to record the trajectories of weakly fluorescent molecules in the nucleus of living cells. I-SPT uses some specific detection and analysis tools to enable the computation of reliable statistics on nuclear particle movement.

• MeSH
• buněčné jádro metabolismus   MeSH
• buněčné linie MeSH
• lidé MeSH
• lidské chromozomy genetika   ultrastruktura   MeSH
• počítačové zpracování obrazu MeSH
• replikace DNA MeSH
• zobrazení jednotlivé molekuly metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

RECQ5 is one of five RecQ helicases found in humans and is thought to participate in homologous DNA recombination by acting as a negative regulator of the recombinase protein RAD51. Here, we use kinetic and single molecule imaging methods to monitor RECQ5 behavior on various nucleoprotein complexes. Our data demonstrate that RECQ5 can act as an ATP-dependent single-stranded DNA (ssDNA) motor protein and can translocate on ssDNA that is bound by replication protein A (RPA). RECQ5 can also translocate on RAD51-coated ssDNA and readily dismantles RAD51-ssDNA filaments. RECQ5 interacts with RAD51 through protein-protein contacts, and disruption of this interface through a RECQ5-F666A mutation reduces translocation velocity by ∼50%. However, RECQ5 readily removes the ATP hydrolysis-deficient mutant RAD51-K133R from ssDNA, suggesting that filament disruption is not coupled to the RAD51 ATP hydrolysis cycle. RECQ5 also readily removes RAD51-I287T, a RAD51 mutant with enhanced ssDNA-binding activity, from ssDNA. Surprisingly, RECQ5 can bind to double-stranded DNA (dsDNA), but it is unable to translocate. Similarly, RECQ5 cannot dismantle RAD51-bound heteroduplex joint molecules. Our results suggest that the roles of RECQ5 in genome maintenance may be regulated in part at the level of substrate specificity.

• MeSH
• adenosintrifosfát metabolismus   MeSH
• bodová mutace MeSH
• helikasy RecQ genetika   metabolismus   ultrastruktura   MeSH
• homologní rekombinace * MeSH
• hydrolýza MeSH
• jednovláknová DNA metabolismus   ultrastruktura   MeSH
• kinetika MeSH
• lidé MeSH
• mikroskopie atomárních sil MeSH
• missense mutace MeSH
• molekulární motory metabolismus   ultrastruktura   MeSH
• rekombinantní fúzní proteiny metabolismus   MeSH
• rekombinantní proteiny metabolismus   MeSH
• rekombinasa Rad51 genetika   metabolismus   MeSH
• replikační protein A metabolismus   MeSH
• substrátová specifita MeSH
• zobrazení jednotlivé molekuly * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

AIM: The aim of this study is to show the current view on fast-track programs optimizing the perioperative care. METHODS: Authors searched Medline databases and identified current trials regarding all factors of fast-track programs. They analyzed these trials and identified the most important principles of fast-track programs based on trial analysis. RESULTS: The most important principles are consistent in all individual trials. Most of authors recommend 10-12 issues. All authors emphasize early mobilization of patients after surgery and early introduction of peroral nutrition. Fast-track programs shorten the duration of hospitalisation and decrease morbidity. CONCLUSION: The perioperative care is characterized by a range of trussing traditions. Fast-track surgery optimizes the perioperative care. It is a safe method according to the trials. The implementation of single fast-track surgery factors is very slow (Tab. 2, Ref. 49).

• MeSH
• délka pobytu MeSH
• lidé MeSH
• perioperační péče metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Background: Super-resolution single molecule localization microscopy (SMLM) is a method for achieving resolution beyond the classical limit in optical microscopes (approx. 200 nm laterally). Yellow fluorescent protein (YFP) has been used for super-resolution single molecule localization microscopy, but less frequently than other fluorescent probes. Working with YFP in SMLM is a challenge because a lower number of photons are emitted per molecule compared with organic dyes, which are more commonly used. Publically available experimental data can facilitate development of new data analysis algorithms. Findings: Four complete, freely available single molecule super-resolution microscopy datasets on YFP-tagged growth factor receptors expressed in a human cell line are presented, including both raw and analyzed data. We report methods for sample preparation, for data acquisition, and for data analysis, as well as examples of the acquired images. We also analyzed the SMLM datasets using a different method: super-resolution optical fluctuation imaging (SOFI). The 2 modes of analysis offer complementary information about the sample. A fifth single molecule super-resolution microscopy dataset acquired with the dye Alexa 532 is included for comparison purposes. Conclusions: This dataset has potential for extensive reuse. Complete raw data from SMLM experiments have typically not been published. The YFP data exhibit low signal-to-noise ratios, making data analysis a challenge. These datasets will be useful to investigators developing their own algorithms for SMLM, SOFI, and related methods. The data will also be useful for researchers investigating growth factor receptors such as ErbB3.

• MeSH
• algoritmy MeSH
• bakteriální proteiny chemie   MeSH
• fluorescenční barviva chemie   MeSH
• lidé MeSH
• luminescentní proteiny chemie   MeSH
• receptory růstových faktorů chemie   izolace a purifikace   MeSH
• zobrazení jednotlivé molekuly metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Advances in fluorescent probe design and synthesis have allowed the uniform in situ labeling of individual RNA molecules. In a technique referred to as single molecule RNA FISH (smRNA FISH), the labeled RNA molecules can be imaged as diffraction-limited spots and counted using image analysis algorithms. Single RNA counting has provided valuable insights into the process of gene regulation. This microscopy-based method has often revealed a high cell-to-cell variability in expression levels, which has in turn led to a growing interest in investigating the biological significance of gene expression noise. Here we describe the application of the smRNA FISH technique to samples of Caenorhabditis elegans, a well-characterized model organism.

• MeSH
• algoritmy MeSH
• Caenorhabditis elegans genetika   růst a vývoj   metabolismus   MeSH
• embryo nesavčí metabolismus   ultrastruktura   MeSH
• fixace tkání metody   MeSH
• fluorescenční barviva chemie   MeSH
• genetická transkripce MeSH
• hybridizace in situ fluorescenční metody   MeSH
• messenger RNA chemie   genetika   metabolismus   MeSH
• poměr signál - šum MeSH
• vývojová regulace genové exprese MeSH
• zobrazení jednotlivé molekuly metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The imaging of healthy tissues and solid tumors benefits from the application of nanoparticle probes with altered pharmacokinetics, not available to low molecular weight compounds. However, the distribution and accumulation of nanoprobes in vivo typically take at least tens of hours to be efficient. For nanoprobes bearing a radioactive label, this is contradictory to the requirement of minimizing the radiation dose for patients by using as-short-as-feasible half-life radionuclides in diagnostics. Thus, we developed a two-stage diagnostic concept for monitoring long-lasting targeting effects with short-lived radioactive labels using bone-mimicking biocompatible polymer-coated and colloidally fully stabilized hydroxyapatite nanoparticles (HAP NPs) and bone-seeking radiopharmaceuticals. Within the pretargeting stage, the nonlabeled nanoparticles are allowed to circulate in the blood. Afterward, 99mTc-1-hydroxyethylidene-1.1-diphosphonate (99mTc-HEDP) is administered intravenously for in situ labeling of the nanoparticles and subsequent single-photon emission computed tomography/computed tomography (SPECT/CT) visualization. The HAP NPs, stabilized with tailored hydrophilic polymers, are not cytotoxic in vitro, as shown by several cell lines. The polymer coating prolongs the circulation of HAP NPs in the blood. The nanoparticles were successfully labeled in vivo with 99mTc-HEDP, 1 and 24 h after injection, and they were visualized by SPECT/CT over time in healthy mice.

• MeSH
• endocytóza MeSH
• fluorescein chemie   MeSH
• fluorescenční barviva chemie   MeSH
• hydroxyapatit chemie   MeSH
• jednofotonová emisní výpočetní tomografie MeSH
• lidé MeSH
• myši inbrední BALB C MeSH
• nádorové buněčné linie MeSH
• nanočástice chemie   ultrastruktura   MeSH
• organotechneciové sloučeniny chemie   MeSH
• počítačová rentgenová tomografie MeSH
• polymery chemie   MeSH
• protonová magnetická rezonanční spektroskopie MeSH
• radiofarmaka chemie   MeSH
• tkáňová distribuce MeSH
• zobrazování trojrozměrné MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: The aim was to compare the speckle tracking echocardiography (STE) derived systolic longitudinal strain (SL(Smax)) with rest single photon emission computed tomography (SPECT) perfusion imaging (Q(REST)), and to define the optimal cut-offs for SL(Smax) to discriminate transmural scar on contrast-enhanced magnetic resonance imaging (ceCMR). METHODS AND RESULTS: In 100 patients with chronic ischemic left ventricular (LV) dysfunction, myocardial viability was assessed using STE and rest SPECT to predict LV segmental relative extent of delayed enhancement (DE) >75% on ceCMR. Correlation was found between regional SL(Smax) (r=-0.59, P<0.0001) and DE on ceCMR. The SL(Smax) optimal cut-off -5.3% identified segments with DE>75% on ceCMR (sensitivity 83.1%, specificity 84.6%). Optimal cut-offs SL(Smax) for segments corresponding to individual perfusion territories (-3.6%, -5.3% and -4.7% for LAD, LCx resp. RCA perfusion territories) were identified. There was a significant difference (AUC 0.866 vs. 0.822 for SL(Smax) resp. Q(REST), p=0.036) in the accuracy of predicting non-viable segment due to the greater accuracy of SL(Smax) than Q(REST) in the RCA perfusion territory (AUC 0.893 vs. 0.75 for SLSmax resp. Q(REST), P=0.001). CONCLUSIONS: STE enabled identification of LV non-viable segments. Cut-off values derived for perfusion territories of individual coronary arteries improve the accuracy of predicting a transmural scar presence. In comparison with rest myocardial SPECT perfusion imaging, STE is more accurate in predicting non-viable myocardium.

• MeSH
• dysfunkce levé srdeční komory diagnóza   MeSH
• echokardiografie * MeSH
• ischemická choroba srdeční diagnóza   patofyziologie   ultrasonografie   MeSH
• jednofotonová emisní výpočetní tomografie * MeSH
• lidé středního věku MeSH
• lidé MeSH
• magnetická rezonanční tomografie MeSH
• odpočinek MeSH
• prediktivní hodnota testů MeSH
• přežití tkáně MeSH
• radiofarmaka diagnostické užití   MeSH
• reprodukovatelnost výsledků MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• zátěžový test MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

RNA-binding proteins (RBPs) are critical to posttranscriptional gene regulation. Therefore, characterization of the RNA molecules bound by RBPs in vivo represent a key step in elucidating their function. The recently developed iCLIP technique allows single nucleotide resolution of the RNA binding footprints of RBPs. We present the iCLIP technique modified for its application to Trypanosoma brucei and most likely other kinetoplastid flagellates. By using the immuno- or affinity purification approach, it was successfully applied to the analysis of several RBPs. Furthermore, we also provide a detailed description of the iCLIP/iCLAP protocol that shall be particularly suitable for the studies of trypanosome RBPs.

• MeSH
• imunoprecipitace metody   MeSH
• nukleotidy genetika   metabolismus   MeSH
• parazitologie metody   MeSH
• proteiny vázající RNA analýza   genetika   metabolismus   MeSH
• protozoální proteiny analýza   genetika   metabolismus   MeSH
• RNA protozoální genetika   metabolismus   MeSH
• RNA genetika   metabolismus   MeSH
• Trypanosoma brucei brucei genetika   MeSH
• ultrafialové záření MeSH
• vazba proteinů genetika   účinky záření   MeSH
• vazebná místa genetika   MeSH
• zobrazení jednotlivé molekuly metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Track structure based simulations valuably complement experimental research on biological effects of ionizing radiation. They provide information at the highest level of detail on initial DNA damage induced by diverse types of radiation. Simulations with the biophysical Monte Carlo code PARTRAC have been used for testing working hypotheses on radiation action mechanisms, for benchmarking other damage codes and as input for modelling subsequent biological processes. To facilitate such applications and in particular to enable extending the simulations to mixed radiation field conditions, we present analytical formulas that capture PARTRAC simulation results on DNA single- and double-strand breaks and their clusters induced in cells irradiated by ions ranging from hydrogen to neon at energies from 0.5 GeV/u down to their stopping. These functions offer a means by which radiation transport codes at the macroscopic scale could easily be extended to predict biological effects, exploiting a large database of results from micro-/nanoscale simulations, without having to deal with the coupling of spatial scales and running full track-structure calculations.

• MeSH
• dvouřetězcové zlomy DNA účinky záření   MeSH
• lidé MeSH
• lineární přenos energie MeSH
• metoda Monte Carlo * MeSH
• poškození DNA * MeSH
• protony * MeSH
• radioterapie * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH