Masaryk University

Improving our understanding of how environmental pollution affects aquatic life requires a holistic approach. This study provides new insights into the intrinsic biological defence of brown trout (Salmo trutta m. fario L.) against chemical pollution in a stream with a low-dilution factor, a common scenario in headwaters globally. Fish restocked downstream of a sewage treatment plant (STP) were compared with a control group upstream of STP. Trout tissues were sampled after 6, 14, and 24 weeks and subjected to biochemical and histological analyses. Passive samplers were deployed at both stream stretches to reflect concentrations of freely dissolved organic micropollutants and their bioactivity effects using in vitro reporter gene bioassays. Chemical analysis downstream revealed elevated concentrations of micropollutants compared to upstream. In vitro bioassays detected increased androgenicity, estrogenicity, and transthyretin-binding inhibition. Antioxidant and biotransformation enzyme activities in fish indicated gradual acclimation to pollution despite minor histopathological changes. Elevated vitellogenin and 17β-estradiol in males suggested pollution-induced endocrine disruption. Although the results obtained from water chemical profiling and bioassays have a causal relationship to fish health, trout's molecular defence system allowed gradual acclimation to pollution, mitigating broader ecological impacts. The study advanced the knowledge of how fish cope with wastewater-borne micropollutants in aquatic environments.

We investigate agreement attraction effects in the L2 English of native speakers of Czech, a language that has little-to-no evidence of attraction effects. Our experiments involve two groups of participants. The first group (N = 415) participated in an L2 English-only experiment, and the second group (N = 183) participated in both L2 English and L1 Czech versions of the experiment (in a randomized order with a two-week interval). Standard attraction effects were observed in L2 English, contrasting with the absence of such effects in L1 Czech. Our results provide unique evidence that an L2 can significantly attract, even when the L1 does not. However, our results also revealed that the attraction effect in L2 English disappeared when the L1 Czech version was completed first. These findings are discussed in relation to the Unified Competition Model and the effects of L2-induced increases in working memory demands.

A series of new Ag(i) and Cd(ii) hydrozanomide-based complexes have been prepared and studied. The synthesized compounds have undergone extensive spectroscopic and structural characterization, including F-AAS, FTIR, single crystal XRD and TGA analyses. Crystal structures of two representative complexes, 4 and 6, have revealed that the silver atom in complex 4 assumes a four-coordinate geometry with two ligands. Additionally, the stability of these complexes in DMSO has been determined using UV-vis spectroscopy. The biological activity of these metal complexes has been assessed, focusing on their antibacterial and anticancer properties against LN-229 and U87 cell lines. Among the tested compounds, silver-based complexes demonstrated significant antibacterial activity against a wide range of microorganisms. Moreover, cytotoxicity studies have shown that the metal complexes exhibit higher anticancer activity compared to their parent ligands. Notably, the Ag(i) complexes of L3, L4, and L5 have emerged as the most promising candidates for their selective toxicity against cancer cells, without harmful effects on normal human fibroblast cells. These findings highlight the potential of Ag(i) complexes as promising anticancer agents with selective toxicity and potent antibacterial properties.

Background For many years, syphilis treatment was considered straightforward due to the universal susceptibility of Treponema pallidum subsp. pallidum (TPA) to penicillin antibiotics. Methods Penicillin-binding protein genes from a ceftriaxone treatment failure T. pallidum isolate were assessed, and the introduction of identified mutations into two laboratory strains via natural competence was aimed for, followed by in vitro analysis of antibiotic susceptibility of the recombinants. Results TPA from the ceftriaxone treatment failure case contained A1873G and G2122A mutations in the TP0705 gene. Introduction of the A1873G mutation into laboratory strains DAL-1 and SS14 resulted in partial resistance to ceftriaxone and penicillin G in vitro. Furthermore, in silico analyses revealed that the majority of contemporary TPA SS14-like strains harbors this mutation and are thus partially resistant to ceftriaxone and penicillin G. Conclusions This finding indicates that TPA strains accumulate mutations that increase their resistance to β-lactam antibiotics. Alternative approaches for controlling syphilis will be needed, including the development of the syphilis vaccine.

The cognitive processing of learning materials has been extensively studied within various cognitive theories. Self-regulated learning (SRL) is also recognized as a key factor in learning efficiency. However, evidence linking SRL to learning outcomes remains inconclusive, particularly regarding objective behavioral data during learning. This study presents an original empirical dataset on eye-tracking activity during learning, examining the effects of metacognitive prompts and multimedia content on cognitive processing and learning outcomes. A controlled laboratory experiment with a 2 × 2 mixed factorial design involved 110 university students, resulting in 84 complete recordings of eye-movement activity during learning. Participants studied scientific materials in text-only and multimedia formats, with one group receiving metacognitive prompts and the control group receiving general instructions. Learning performance was assessed via a post-test, and eye-tracking technology captured gaze patterns to provide insights into cognitive engagement and attention distribution. Applications extend to e-learning, virtual environments, and user interface design. While the dataset has some methodological limitations, it remains a robust resource for studying cognitive processes and optimizing educational technologies.

The development of vaccines has significantly advanced public health by preventing infectious diseases. However, traditional vaccines face challenges such as the need for injections, trained personnel, and cold storage. Plant-derived edible vaccines present a promising alternative, offering cost-effectiveness, scalability, and the ability to induce mucosal immunity. Duckweeds (Lemnaceae), with their high protein content and rapid growth, have received interest in producing these vaccines. Transgenic duckweed expressing pathogenic antigens has demonstrated efficacy in animal models, eliciting both mucosal and systemic immune responses. Despite their potential, large-scale production of duckweed-based vaccines faces challenges, including spatial requirements, water usage, and contamination risks. To address these issues, we propose a vertical farming and recirculation system. This system minimizes space and water usage, enhances light exposure through transparent, retractable trays, and integrates solar panels for energy efficiency. Additionally, utilizing livestock and poultry farm wastewater as a growth medium offers a sustainable solution. Our proposed value chain involves transforming duckweed with molecular bioengineering techniques, cultivating it vertically, and processing it into a powder for food or feed. This approach promises to enhance vaccine accessibility by lowering storage requirements, as the powdered biomass does not need to be refrigerated. This could reduce costs and contribute to global public health efforts. Still, purification steps before vaccine usage and potential regulatory concerns are yet to be addressed.

Objective: Accurate localization of the epileptogenic zone (EZ) is crucial for epilepsy surgery, but the class imbalance of epileptogenic vs. normal electrode contacts in intracranial EEG (iEEG) data poses significant challenges for automatic localization methods. This review evaluates methodologies for handling the class imbalance in EZ localization studies that use machine learning. Approach: We systematically reviewed studies employing machine learning to localize the EZ from iEEG data, focusing on strategies for addressing class imbalance in data handling, algorithm design, and evaluation. Results: Out of 2,128 screened studies, 35 fulfilled the inclusion criteria. Across the studies, the iEEG contacts identified as epileptogenic by the localization algorithm constituted a median of 18.34% of all contacts. However, many of these studies did not adequately address the class imbalance problem. Techniques such as data resampling and cost-sensitive learning were used to mitigate the class imbalance problem, but the chosen evaluation metrics often failed to account for it. Significance: Class imbalance significantly impacts the reliability of EZ localization models. More comprehensive management and innovative approaches are needed to enhance the robustness and clinical utility of these models. Addressing class imbalance in machine learning models for EZ localization will improve both the predictive performance and reliability of these models.

Background Performance-based risk-sharing agreements (PBRSA) represent an innovative tool for managing uncertainty and balanced distribution of the financial risk of high-cost drugs. By linking reimbursement to real-world treatment performance, these agreements help mitigate budgetary impacts. This study poses an illustrative patient-level PBRSA reimbursement model for non-small cell lung cancer (NSCLC) immunotherapy based on collected real-world data (RWD). Methods A retrospective analysis of 266 patients with NSCLC treated with immunotherapy was performed. Progression-free survival (PFS) served as the primary outcome measure of therapeutic effectiveness. An illustrative patient-level PBRSA model was developed to quantify the manufacturer’s financial participation based on deviations from established PFS thresholds reported in randomised controlled trials (RCT). The manufacturer’s financial responsibility increased proportionally to greater deviations in patient outcomes from the RCT benchmark. Cost calculations were limited exclusively to the acquisition price of immunotherapies, excluding administration, toxicity management, and other indirect costs. The potential PBRSA scenario was compared with the current reimbursement situation. Results Using this reimbursement method, cost savings per checkpoint inhibitor for healthcare payers could represent between 27.3% and 66.2% of the total cost, depending on the individual PFS reached. For the RWD cohort of NSCLC patients unsuccessfully treated with pembrolizumab monotherapy was 57.5% (a reduction in cost to payers from $27 996 to$11 893 per patient); pembrolizumab in combination 51.7% ($33 595 to$16 237); nivolumab 37.1% ($5 608 to$3 531); atezolizumab 27.3% ($11 799 to$8 583); and durvalumab 66.2% ($44 005 to$14 882). Conclusions This study proposes an illustrative patient-level PBRSA reimbursement model leveraging real-world clinical data to enhance risk-sharing for high-cost therapies. Unlike conventional cost-effectiveness analyses, this method directly links clinical performance and manufacturer financial responsibility. Future research should integrate comprehensive cost considerations and validate model performance in broader clinical settings.

Background The online environment provides adolescents with vast amounts of health-related information; however, navigating this effectively requires high levels of eHealth literacy to avoid misinformation and harmful content. Parental guidance is often considered a crucial factor in shaping adolescents’ online health behaviors; however, there is limited longitudinal research examining how parental eHealth literacy mediation influences adolescents’ development of eHealth literacy over time. Objective This study aims to examine the reciprocal relationship between parental eHealth literacy mediation and adolescents’ eHealth literacy. It also investigates whether parental education moderates this relationship, specifically exploring whether higher levels of parental education enhance the effectiveness of eHealth literacy mediation in improving adolescents’ eHealth literacy. Methods A 3-wave longitudinal study was conducted, collecting data from 2500 adolescent-parent pairs. A random intercept cross-lagged panel model was applied to assess the reciprocal effects between parental eHealth literacy mediation and adolescents’ eHealth literacy across the 3 waves. Parental education was included in the model as a potential moderating variable to examine whether it influences the strength of the relationship between parental eHealth literacy mediation and adolescents’ eHealth literacy. Results The findings revealed no significant within-person effects, indicating that changes in parental eHealth literacy mediation over time did not lead to corresponding changes in adolescents’ eHealth literacy (T1→T2 β=–.03, P=.65; T2→T3 β=.01, P=.84), and vice versa (T1→T2 β=.02, P=.71; T2→T3 β=–.07, P=.19). Furthermore, the data did not support a moderating effect of parental education, suggesting that higher educational attainment does not enhance the impact of parental eHealth literacy mediation. However, a significant between-person association was observed: adolescents with higher levels of eHealth literacy tend to have parents who engage more frequently in eHealth literacy mediation (r=0.30, P<.001). Conclusions This study contributes to the understanding of parental involvement in shaping adolescents’ eHealth literacy. Contrary to expectations, parental eHealth literacy mediation does not appear to have a significant longitudinal impact on the development of adolescents’ eHealth literacy, nor does higher parental education strengthen this relationship. These findings suggest that additional factors beyond parental mediation and education may play a critical role in supporting adolescents’ ability to navigate online health information effectively.

Background Exposure to the environments with limited walkability and high density of unhealthy food outlets promotes obesity development and might cluster in disadvantaged neighbourhoods. This study examines the combined obesogenicity of urban neighbourhoods in Brno, and related socio-economic disparities. Methods This study was conducted in Brno, the second-largest city in Czechia. The obesogenic index was calculated from 12 components of built food and physical activity environments for each of the 296 basic settlement units (BSUs) of Brno. The index ranged from 0 (low obesogenicity) to 100 (high obesogenicity). The social disparities were assessed using linear regression. Spatial clustering was assessed using the global Moran’s Index. Results The median obesogenic index score for Brno’s 296 BSUs was 72.09 (IQR = 24.03), with higher scores in peripheral and industrial areas. Areas with higher proportion of people with university education had lower obesogenic scores of physical activity and overall obesogenic environment. Simultaneously, localities with higher unemployment exhibited lower obesogenic score in food and overall obesogenic environment. Conclusion Areas with lower levels of obesogenicity were primarily concentrated in central locations. No clear socio-economic gradient was observed, although proportion of university-educated inhabitants and unemployment rates were both associated with lower obesogenic environment scores.

The effectiveness of assisted reproduction methods is still not as high as was expected recently. For effective therapy, it is necessary to use only the highest quality sperm for oocyte fertilization, as the fertilization process most often takes place using the ICSI technique, i.e. direct injection of sperm into the oocyte. For these reasons, separation techniques are constantly being developed, and in a relatively short period of time the view of these procedures has changed. In recent years, microfluidic chip techniques have been increasingly applied. In this study, we compared the swim up technique with the two most used chips (ca0 and ZYMOT). We tested the efficiency of these separation methods against the swim-up method. In this study, we evaluated sperm concentration, motility, morphology, acrosome status and DNA integrity before and after separation methods. Each patient’s ejaculate was separated simultaneously by the swim up method and both chips. It turned out that the separation methods are very similar in motility, concentration and acrosome status and sperm morphology. However, in the case of DNA fragmentation, a significant difference in the reduction of the proportion of fragmented spermatozoa was found only with the chips and not with the swim-up method. Moreover, when comparing the relative effectiveness of sperm reduction with fragmented DNA, the ZYMOT method was more effective than Ca0. However, the differences between the chips were not statistically significant with an average of 18%.

Chloroplasts are the sites of photosynthesis but also host essential metabolic and biosynthetic pathways; therefore, the regulation of the chloroplast population is of crucial importance for the viability of the plant. Chloroplast division is closely linked to leaf development, but the coordination of cell expansion, division, and chloroplast multiplication at the molecular level is still poorly understood. Auxin signalling influences leaf growth and may also mediate chloroplast biogenesis and proliferation. Most studies focused on auxin and the development of chloroplasts in the fruit, emphasising the need for further research on leaf tissue. Overexpression of Growth Regulating Factor 5 (35S:GRF5) increases cell and chloroplast division in Arabidopsis thaliana, resulting in larger leaves with more chloroplasts per cell. In this study, we utilised 35S:GRF5 plants as a model to identify auxin control points that regulate chloroplast division. By examining the impact of changes in auxin homeostasis on chloroplast division and mesophyll cell size and by analysing crosses with selected auxin homeostasis genes, we found that reactive oxygen species‐auxin crosstalk influences chloroplast multiplication during the cell expansion phase. Evidence indicates that GRF5 modulates auxin responsiveness by regulating the expression of UDP‐glucosyltransferase UGT74E2 and PIN‐LIKES3/5, which are key players in intracellular auxin homeostasis. These findings provide potential targets for modulating chloroplast abundance to improve photosynthetic efficiency in crops and highlight key areas for further research.

This study investigates the hydration reaction of a gold(I)–N-heterocyclic carbene [Au(I)–NHC] complex at both the quantum mechanical (QM) level and combined Quantum Mechanics/Molecular Mechanics (QM/MM) MD simulations. The main goals are to analyze the differences between implicit (PCM) and explicit solvation models and to compare the advantages and disadvantages of both approaches. Regarding the QM part, the B97D3 and B3PW91 functionals are combined with double-zeta basis sets and the C-PCM/UFF implicit solvation model and compared with the CCSD(T)/TZP computational level supplemented with the C-PCM (COSMO/Klamt radii) or D-PCM/scaled-UAKS solvation model. In addition, reaction force and reaction electronic flux (REF) analyses are performed along the intrinsic reaction coordinate (IRC) determined at the B3PW91/6-31+G(d)/SDD/C-PCM/UFF computational model for deeper insights into the reaction mechanism. Despite relatively high endergonicity, the TS structure is quite close to the center of the reaction coordinate, contrary to the Hammond principle. In the QM/MM MD part of the study, the B97D3 computational setting from the previous part is used as a QM core, and several different explicit water solvation models are explored in the MM environment. The TIP3P water model is compared with the OPC, POL3, TIP4P, and SPCE ones. Nevertheless, they all lead to very low activation barriers and mild endergonicity. Both ΔGr and ΔGa energies are visibly reduced compared to QM values when PCM models are applied. Since partial charges of water atoms within the QM calculations are visibly smaller than point charges in all the explored force-field water models, a modified TIP3P (with partial charges close to DFT RESP values and a LJ parameter conserving the correct water density) is used. In this manner, the energy profile is closer to QM results (with ΔGa = 8.2 and ΔGr = 6.4 kcal mol⁻¹)—especially to the CCSD(T)/TZP/D-PCM/scaled-UAKS model (ΔGa = 14.6 and ΔGr = 9.1). Nevertheless, the hydration process is predicted to be endoergic in all explored models.

Cilia are versatile, microtubule-based organelles that facilitate cellular signaling, motility, and environmental sensing in eukaryotic cells. These dynamic structures act as hubs for key developmental signaling pathways, while their assembly and disassembly are intricately regulated along cell cycle transitions. Recent findings show that factors regulating ciliogenesis and cilia dynamics often integrate their roles across other cellular processes, including cell cycle regulation, cytoskeletal organization, and intracellular trafficking, ensuring multilevel crosstalk of mechanisms controlling organogenesis. Disruptions in these shared regulators lead to broad defects associated with both ciliopathies and cancer. This review explores the crosstalk of regulatory mechanisms governing cilia assembly, disassembly, and maintenance during ciliary signaling and the cell cycle, along with the broader implications for development, tissue homeostasis, and disease.

The paper examines changes in the habits and customs of visitors of the Barum Czech Rally Zlín before and after the COVID-19 pandemic. Through a survey, we found that while overall habits remained largely unchanged, several changes were observed. Post-covid, there was a higher reliance on car transportation, longer stays at the event, increased use of hotels, reduced restaurant dining, and greater intention to revisit the region. Findings indicate the pandemic's influence on transportation choices, accommodation preferences, and increased loyalty to the rally event. This research focuses solely on rally visitors, making the results specific to this niche group.

Let $\Omega \subset {\mathbb {R}}^N$ ($N \ge 3$) be a $C^2$ bounded domain and $\Sigma \subset \Omega$ is a $C^2$ compact boundaryless submanifold in ${\mathbb {R}}^N$ of dimension k, $0\le k < N-2$. For $\mu \le (\frac{N-k-2}{2})^2$, put $L_\mu := \Delta + \mu d_{\Sigma }^{-2}$ where $d_{\Sigma }(x) = \textrm{dist}(x,\Sigma )$. We study boundary value problems for equation $-L_\mu u = g(u,|\nabla u|)$ in $\Omega \setminus \Sigma$, subject to the boundary condition $u=\nu$ on $\partial \Omega \cup \Sigma$, where $g: {\mathbb {R}} \times {\mathbb {R}}_+ \rightarrow {\mathbb {R}}_+$ is a continuous and nondecreasing function with g(0,0)=0, $\nu$ is a given nonnegative measure on $\partial \Omega \cup \Sigma$. When g satisfies a so-called subcritical integral condition, we establish an existence result for the problem under a smallness assumption on $\nu$. If $g(u,|\nabla u|) = |u|^p|\nabla u|^q$, there are ranges of p, q, called subcritical ranges, for which the subcritical integral condition is satisfied, hence the problem admits a solution. Beyond these ranges, where the subcritical integral condition may be violated, we establish various criteria on $\nu$ for the existence of a solution to the problem expressed in terms of appropriate Bessel capacities.

DNA methylation-based classification of (brain) tumors has emerged as a powerful and indispensable diagnostic technique. Initial implementations used methylation microarrays for data generation, while most current classifiers rely on a fixed methylation feature space. This makes them incompatible with other platforms, especially different flavors of DNA sequencing. Here, we describe crossNN, a neural network-based machine learning framework that can accurately classify tumors using sparse methylomes obtained on different platforms and with different epigenome coverage and sequencing depth. It outperforms other deep and conventional machine learning models regarding accuracy and computational requirements while still being explainable. We use crossNN to train a pan-cancer classifier that can discriminate more than 170 tumor types across all organ sites. Validation in more than 5,000 tumors profiled on different platforms, including nanopore and targeted bisulfite sequencing, demonstrates its robustness and scalability with 99.1% and 97.8% precision for the brain tumor and pan-cancer models, respectively.

The ADARB1 gene encodes the ADAR2 RNA editing enzyme, which edits the GRIA2 transcript Q/R editing site with almost 100% efficiency in the nervous system. The edited GRIA2R transcript encodes the GLUA2 R subunit isoform of tetrameric AMPA receptors, which is essential to prevent seizures associated with aberrantly elevated AMPA receptor cation permeability. Rare biallelic variants in ADARB1 cause severe infant and childhood seizures and developmental delays in seven cases we previously described. Here, we report two new homozygous ADARB1 variants and study ADAR2 variant editing activities at the GRIA2 Q/R site and other editing sites in cell cultures. One new variant in the second dsRNA binding domain (dsRBD II) retains up to 60% editing activity, whereas another, in the deaminase domain, eliminates RNA editing activity. Reduced GRIA2 Q/R site editing increases AMPA receptor permeability by upregulating the expression of the GLUA2 Q isoform and reducing overall GLUA2 subunit levels, resulting in AMPA receptors that lack GLUA2 and are calcium-permeable. Since failure to edit the GRIA2 Q/R site leads to failure of intron 11 splicing, we also examined the effects of ADAR2 variants on the splicing of a mouse Gria2-based reporter and concluded that ADAR2 variants affect splicing only through their effects on RNA editing activity. To expand the number of variants in ADARB1, some variants reported in ClinVar have also been analysed by in silico methods to predict which are likely to be most deleterious and associated with seizures in patients.