During hypoxia, tissues are subjected to an inadequate oxygen supply, disrupting the balance needed to maintain normal function. This deficiency can occur due to reduced oxygen delivery caused by impaired blood flow or a decline in the blood's ability to carry oxygen. In tumors, hypoxia and vascularization play crucial roles, shaping their microenvironments and influencing cancer progression, response to treatment and metastatic potential. This chapter provides guidance on the use of non-invasive imaging methods including Positron Emission Tomography and Magnetic Resonance Imaging to study tumor oxygenation in pre-clinical settings. These imaging techniques offer valuable insights into tumor vascularity and oxygen levels, aiding in understanding tumor behavior and treatment effects. For example, PET imaging uses tracers such as [18F]-fluoromisonidazole (FMISO) to visualize hypoxic areas within tumors, while MRI complements this with anatomical and functional images. Although directly assessing tumor hypoxia with MRI remains challenging, techniques like Blood Oxygen Level Dependent (BOLD) and Dynamic Contrast-Enhanced MRI (DCE-MRI) provide valuable information. BOLD can track changes in oxygen levels during oxygen challenges, while DCE-MRI offers real-time access to perfusion and vessel permeability data. Integrating data from these imaging modalities can help assess oxygen supply, refine treatment strategies, enhance therapeutic effectiveness, and ultimately improve patient outcomes.

• Klíčová slova
• BOLD, DCE-MRI, FMISO, Hypoxia, Magnetic resonance imaging, Positron emission tomography, Preclinical, Tumor oxygenation, Vascularity,
• MeSH
• hypoxie diagnostické zobrazování   MeSH
• kyslík metabolismus   MeSH
• lidé MeSH
• magnetická rezonanční tomografie * metody   MeSH
• misonidazol analogy a deriváty   MeSH
• myši MeSH
• nádorová hypoxie MeSH
• nádory diagnostické zobrazování   krevní zásobení   patologie   MeSH
• patologická angiogeneze diagnostické zobrazování   patologie   MeSH
• pozitronová emisní tomografie * metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fluoromisonidazole MeSH Prohlížeč
• kyslík MeSH
• misonidazol MeSH

The analysis of microplastics with current spectroscopic and pyrolytic methods is reaching its limits, especially with regard to detailed spatial distribution in biological tissues. This limitation hampers a comprehensive understanding of the effects of microplastics on organisms. Therefore, there is a pressing need to expand the analytical approaches to study microplastics in biota. In this context, the aim of this study was to test the applicability of non-destructive 3D imaging using X-ray micro-computed tomography (microCT) for the detection of microplastics in fish. Zebrafish (Danio rerio) were gavaged with polyethylene spherical microplastics (30-110 μm) and the distribution of microplastics in the gut was investigated using microCT. The results showed that the particle size distribution determined by microCT closely matched the data from conventional laser diffraction analysis. In addition, microCT was able to detect microplastics in spiked fish tissue and provide precise localization data by tracing particles of known type and shape. MicroCT offers a novel approach for tracking microplastics in organisms and enables accurate sizing without compromising the integrity of the tissue under investigation. It therefore represents a valuable addition to spectroscopic methods, which are widely used for the detection of microplastics based on their chemical composition but do not provide data on their spatial distribution.

• Klíčová slova
• Environment, Imaging, Microplastics, Plastic Pollution, X-ray Computed Tomography,
• MeSH
• chemické látky znečišťující vodu * analýza   MeSH
• dánio pruhované * metabolismus   MeSH
• mikroplasty * analýza   MeSH
• polyethylen chemie   MeSH
• rentgenová mikrotomografie * MeSH
• velikost částic MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chemické látky znečišťující vodu * MeSH
• mikroplasty * MeSH
• polyethylen MeSH

OBJECTIVES: Chemical cross-linking coupled with mass spectrometry (CXMS) offers the distance constraints critical for building the structural model of protein and protein complexes and understanding dynamics of biological systems. Originally developed for protein structural models, CXMS has evolved into method for studying protein complex formation, ligand-induced conformational changes, and quantitative structural analysis using isotopically labeled cross-linkers. METHODS: In this study we tested the potential of isotopically labeled MS-cleavable cross-linker to track the stability of the therapeutic monoclonal antibodies. RESULTS: A novel isotopically labeled MS-cleavable cross-linker was synthesized, and its reactivity was successfully tested on peptide and protein standards. Further, the novel cross-linker was utilized to test the stability of selected therapeutical monoclonal antibodies, bevacizumab and trastuzumab, adopting the data-independent acquisition. CONCLUSION: This study reports the advantages of using combination of 13C isotopically labeled MS-cleavable cross-linkers and data-independent mass spectrometry analysis for the automated identification of cross-linked products and thus monitoring the structural rearrangement of protein structure.

• Klíčová slova
• Chemical cross-linking, antibody, data-independent analysis, mass spectrometry, protein structure, stable isotopes,
• Publikační typ
• časopisecké články MeSH

The COVID-19 pandemic placed significant psychological strain on healthcare workers. Our study tracked health care workers in the Czech Republic throughout the pandemic to examine the impact of stressors on psychological distress over time and explore gender differences in these associations. We studied health care workers from the Czech arm of the international COVID-19 HEROES Study who took part in all three waves of data collection in 2020, 2021 and 2022 (n = 264). We employed a path model to examine the relationships among six stressors (low trust in workplace, assignment of new tasks, prioritization of patients, experience of death due to COVID-19, experience of discrimination or violence, contact with COVID-19 patients), psychological distress (measured by General Health Questionnaire), and covariates (age, gender, occupation). We incorporated autoregressive paths and interactions to assess the longitudinal impact of stressors. Prior levels of distress predicted subsequent distress, with significant carry-over effects observed between each wave. Stressors had direct association to distress in the same wave of data collection (2020 and 2021), but not in subsequent waves, and stressors in 2022 did not have a significant direct effect to distress. Men reported lower distress in 2020 and 2021, but not in 2022. Furthermore, no significant interactions between stressors and gender were found. Age and occupation were not related to distress levels. There is a need for adaptable mental health support that addresses current stressors through targeted interventions, while also providing ongoing monitoring beyond crises, particularly for individuals with high distress levels.

• Klíčová slova
• COVID-19, Distress, Health care worker, Occupational stressors, Pandemic,
• MeSH
• COVID-19 * psychologie   epidemiologie   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• pandemie MeSH
• pracovní stres psychologie   MeSH
• průzkumy a dotazníky MeSH
• psychický distres * MeSH
• psychický stres * epidemiologie   MeSH
• SARS-CoV-2 MeSH
• zdravotnický personál * psychologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

Given the increased interest in incorporating linear energy transfer (LET) as an optimization parameter in intensity-modulated proton therapy (IMPT), a solution for experimental validation of simulations and patient-specific quality assurance in terms of proton LET is needed. Here, we present the methodology and results of LET spectra measurements for spread-out Bragg peak (SOBP) and IMPT plans using a miniaturized pixel detector Timepix3. Approach: We used a MiniPIX Timepix3 detector that provides single-particle tracking, type-resolving power, and spectral information while allowing measurement in quasi-continuous mode. We performed measurements for SOBP and IMPT plans in homogeneous RW3 and heterogeneous CIRS head phantoms with reduced beam current. An artificial intelligence-based model was applied for proton identification and a GPU-accelerated FRED Monte Carlo (MC) code was applied for corresponding MC simulations. Main results: We compared the deposited energy and LET spectra obtained in mixed radiation fields from measurements and MC simulations. The peak positions of deposited energy and LET spectra for the SOBP and IMPT plans agree within the error bars. Discrepancies exceeding the error bars are only visible in the logarithmic scale in high-energy deposition and high-LET tails of the distributions. The mean relative difference of dose-averaged LET values between measurements and MC simulations for individual energy layers is about 5.1%. Significance: This study presents a methodology for assessing radiation quality in proton therapy through energy deposition and LET spectra measurements in uniform and clinical IMPT fields. Findings show an agreement between experimental data and MC simulations, validating our approach. The presented results demonstrate the feasibility of a commercially available Timepix3 detector to validate LET computations in IMPT fields and perform patient-specific quality assurance in terms of LET. This will support the implementation of LET in treatment planning, which will ultimately increase the effectiveness of the treatment. .

• Klíčová slova
• intensity-modulated proton therapy, linear energy transfer, semiconductor pixel detector,
• Publikační typ
• časopisecké články MeSH

Strict medication adherence, which reflects the process by which patients take their medication as prescribed, is crucial for the use of direct oral anticoagulants (DOACs). Therefore, technological devices may serve as promising tools for assessing adherence. We aimed to systematically review the literature focusing on electronically monitored adherence (EMA) to DOACs. All studies indexed in EMBASE, Cochrane Library, MEDLINE, Scopus, and Web of Science from inception until September 1, 2023, were searched. Original studies targeting the query topics were included, findings were categorized and narratively synthetized. Adherence data, including the quality of data reporting bias, were evaluated using the EMERGE guideline. The review protocol was registered in the PROSPERO database (ID CRD42023441161). Out of the 5911 potential hits, 19 articles, comprising 15 research studies, were identified. These studies enrolled 4163 patients (median 72.1 years; 57.9% males), usually chronically treated with DOACs for atrial fibrillation. EMA was measured in 3451 patients by seven different devices from eight manufacturers; the median population tracked with electronic monitoring was 56 patients over 5 months per study. Observational studies resulted in 88.6% and interventional studies resulted in 92.5% of EMA to DOACs, mostly monitoring regimen and taking adherence. Two studies reported high-quality adherence data, whereas 11 reported low-quality adherence data. The item described in the EMERGE guideline as affecting adherence by measurement method, as appropriate, has rarely been addressed. This review broadens the understanding of the overall high EMA to DOACs reported across various study populations and designs. Furthermore, due to the identified gaps in current literature, it highlights the pressing need for standardized methodologies and improved adherence reporting. This study was supported by the GAUK 328322 and SVV 220665.

• Klíčová slova
• anticoagulants, atrial fibrillation, bias, electronics, implementation, medication adherence,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

INTRODUCTION: The functional programs of CD4+ T helper (Th) cell clones play a central role in shaping immune responses to different challenges. While advances in single-cell RNA sequencing (scRNA-Seq) have significantly improved our understanding of the diversity of Th cells, the relationship between scRNA-Seq clusters and the traditionally characterized Th subsets remains ambiguous. METHODS: In this study, we introduce TCR-Track, a method leveraging immune repertoire data to map phenotypically sorted Th subsets onto scRNA-Seq profiles. RESULTS AND DISCUSSION: This approach accurately positions the Th1, Th1-17, Th17, Th22, Th2a, Th2, T follicular helper (Tfh), and regulatory T-cell (Treg) subsets, outperforming mapping based on CITE-Seq. Remarkably, the mapping is tightly focused on specific scRNA-Seq clusters, despite 4-year interval between subset sorting and the effector CD4+ scRNA-Seq experiment. These findings highlight the intrinsic program stability of Th clones circulating in peripheral blood. Repertoire overlap analysis at the scRNA-Seq level confirms that the circulating Th1, Th2, Th2a, Th17, Th22, and Treg subsets are clonally independent. However, a significant clonal overlap between the Th1 and cytotoxic CD4+ T-cell clusters suggests that cytotoxic CD4+ T cells differentiate from Th1 clones. In addition, this study resolves a longstanding ambiguity: we demonstrate that, while CCR10+ Th cells align with a specific Th22 scRNA-Seq cluster, CCR10-CCR6+CXCR3-CCR4+ cells, typically classified as Th17, represent a mixture of bona fide Th17 cells and clonally unrelated CCR10low Th22 cells. The clear distinction between the Th17 and Th22 subsets should influence the development of vaccine- and T-cell-based therapies. Furthermore, we show that severe acute SARS-CoV-2 infection induces systemic type 1 interferon (IFN) activation of naive Th cells. An increased proportion of effector IFN-induced Th cells is associated with a moderate course of the disease but remains low in critical COVID-19 cases. Using integrated scRNA-Seq, TCR-Track, and CITE-Seq data from 122 donors, we provide a comprehensive Th scRNA-Seq reference that should facilitate further investigation of Th subsets in fundamental and clinical studies.

• Klíčová slova
• T cell memory, Th17, Th22, cytotoxic CD4+ T cells, helper T cell subsets, immune repertoires, scRNA-Seq, scTCR-seq,
• MeSH
• analýza genové exprese jednotlivých buněk MeSH
• analýza jednotlivých buněk * metody   MeSH
• lidé MeSH
• sekvenční analýza RNA MeSH
• sekvenování transkriptomu * metody   MeSH
• T-lymfocyty - podskupiny * imunologie   MeSH
• T-lymfocyty pomocné-indukující * imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Anthropogenic activities threaten many wildlife populations by increasing mortality rates, making it crucial to identify the locations and causes of mortality to inform conservation actions. Technological advancements, such as GPS satellite tracking, enable precise recording of wildlife movements. High-resolution data from such devices can facilitate rapid carcass recovery and provide insights into the mortality causes of tagged individuals. Obtaining required information to determine these causes is complex, and standardized approaches can overcome these limitations. In this study, we introduce the LIFE EUROKITE Assessment Protocol (LEAP), a framework for determining the timing, locations, and causes of mortality in GPS-tagged birds. LEAP is a multifaceted approach that integrates: (1) GPS tracking data, (2) evidence from the mortality location (site investigation), and (3) necropsy results to derive the mortality cause and a corresponding certainty score. We supplement the detailed description of LEAP with case studies assessing its effectiveness. Using 329 deceased GPS-tagged red kites (Milvus milvus) we compared conditions of the carcasses processed using LEAP with 145 opportunistically collected raptor carcasses. We also show that LEAP improves carcass condition and therefore allows for higher quality necropsy results. Additionally, we assessed how availability among sources of information (tracking, site investigation and necropsy) influences the quality of mortality assessments. Applying LEAP with all data sources provided the highest quality assessments in 64% of cases. Some 35% of cases were of high quality without necropsy, instead drawing evidence only from tracking data and site investigations. Predation related mortality was less prevalent (11%) when relying on necropsy compared to cases without necropsy (36%), while poisoning showed the opposite trend. Furthermore, we provide guidelines and empirical examples of mortality assessments. Our standardized LEAP approach ensures the best use of all available information regarding mortality events in GPS-tagged birds and advances wildlife mortality research as a valuable tool for conservationists and wildlife managers.

• Klíčová slova
• GPS tracking, bird crime, cause of death, human‐wildlife conflict, population monitoring, survival analysis, wildlife conservation,
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: The objective of this study was to assess the impact of the Enhanced Recovery After Surgery (ERAS) programme implementation on treatment costs at a university-type centre, using the DRG scheme. MATERIALS AND METHODS: Retrospective analysis of patients' data in a group of 604 individuals enroled in the study. We evaluated three groups of patients according to the ERAS clinical protocol (CP): (1) CP oncogynaecology, (2) CP simple hysterectomy, (3) CP laparoscopy. The study aimed to evaluate the impact on the length of stay (LOS), savings in bed-days, and the reduction in direct treatment costs. Three parameters-antibiotic consumption, blood derivative consumption and laboratory test costs-were chosen to compare direct treatment costs. The statistical significance of the difference in the observed parameters was tested by a two-sample unpaired t test with unequal variances at the 0.05 significance level. RESULTS: We analysed data from 604 patients. In all three groups, the length of stay (LOS) was significantly reduced. The most significant reduction was observed in the CP oncogynaecology group, where the LOS was reduced from 11.1 days to 6.8 days (2022) and 7.6 days (2023) compared to 2019 (p < 0.05). Furthermore, there was a notable reduction in inpatient bed-days, which resulted in the capacity being made available to admit additional patients. A statistically significant reduction in direct costs was observed in the group of CP hysterectomy (antibiotic use) and in the CP laparoscopy (laboratory test costs). CONCLUSIONS: The implementation of the ERAS principles resulted in a number of significant positive economic impacts-reduction in the LOS and a corresponding increase in bed capacity for new patients. Additionally, direct treatment costs, including those related to antibiotic use or laboratory testing were reduced. The Czech Republic's acute healthcare system, like the majority of European healthcare systems, is financed by the DRG system. This flat-rate payment per patient encourages hospital management to seek cost-reduction strategies. The results of our study indicate that fast-track protocols represent a potential viable approach to reducing the cost of treatment while simultaneously meeting the recommendations of evidence-based medicine.

• Klíčová slova
• diagnosis related groups, enhanced recovery after surgery, gynaecologic oncology, gynaecologic surgical procedures, health care costs, outcome assessment,
• MeSH
• délka pobytu statistika a číselné údaje   ekonomika   MeSH
• dospělí MeSH
• gynekologické chirurgické výkony * ekonomika   MeSH
• hysterektomie ekonomika   MeSH
• klasifikační systém DRG MeSH
• laparoskopie ekonomika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• náklady na zdravotní péči * statistika a číselné údaje   MeSH
• retrospektivní studie MeSH
• senioři MeSH
• urychlená pooperační rehabilitace * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The hippocampus (HPC) is essential for navigation and memory, tracking environmental continuity and change, including navigation relative to moving targets. CA1 ensembles expressing immediate-early gene (IEG) Arc and Homer1a RNA are contextually specific. While IEG expression correlates with HPC-dependent task demands, the effects of behavioral demands on IEG-expressing ensembles remain unclear. In three experiments, we investigated the effects of context switch, sustained presence, and task demands on dorso-proximal CA1 IEG+ ensembles in rats. Experiment 1 showed that the size of IEG+ (Arc, Homer1a RNA) ensembles dropped to baseline during uninterrupted 30-min exploration, reflecting familiarization, unless a context switch was present. Context-specificity of the ensembles depended on both environment identity and timing of the context switch. Experiment 2 found no effect of HPC-dependent mobile robot avoidance or HPC-independent avoidance of a stationary robot on IEG+ ensembles beyond mere exploration. Experiment 3 replicated these findings for c-Fos protein. The data suggest that IEG+ ensembles are driven by a context switch and shrink over time during sustained presence in the same environment. We found no evidence of task demands or their change affecting the size, stability over time, or task-specificity of IEG+ ensembles. These results shed light on the temporal dynamics of CA1 IEG+ ensembles, and their control by contextual and behavioral factors.

• Klíčová slova
• Arc/Homer1a catFISH, Behavioral episode, C-Fos, Ensemble stability, Moving/stationary robot avoidance, RNA,
• MeSH
• chování zvířat fyziologie   MeSH
• cytoskeletální proteiny genetika   metabolismus   MeSH
• hipokampální oblast CA1 * metabolismus   fyziologie   MeSH
• Homer scaffold proteiny * metabolismus   genetika   MeSH
• krysa rodu Rattus MeSH
• okamžité časné geny * fyziologie   MeSH
• potkani Long-Evans * MeSH
• proteiny nervové tkáně genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• activity regulated cytoskeletal-associated protein MeSH Prohlížeč
• cytoskeletální proteiny MeSH
• Homer scaffold proteiny * MeSH
• Homer1 protein, rat MeSH Prohlížeč
• proteiny nervové tkáně MeSH