Application of passive sampling in monitoring of pyrethroids in surface water and wastewater
Adam Březina

Pyrethroids are widely used synthetic insecticides that are highly hydrophobic and toxic to aquatic organisms at very low concentrations, making their monitoring analytically challenging. This study investigated the occurrence and distribution of selected pyrethroids in the Danube River and municipal wastewater using passive sampling within Joint Danube Survey 5 in summer 2025. Silicone-based passive samplers were deployed at nine surface water sites and nine wastewater treatment plant effluents. Pyrethroids were detected by GC–(APCI)-QqQ in samples, with concentrations ranging from 14.1–2000 pg L⁻¹ in surface waters and 502–6430 pg L⁻¹ in effluents. Annual average environmental quality standards for pyrethroids, according to European Directive 2026/805, were exceeded at four surface water sites, mainly due to deltamethrin.

Spatial Patterns of Parkinson's Disease Prevalence: The Need for High-Resolution Spatial Epidemiology
Aneta Vrzalová

Using individual-level Parkinson’s disease (PD) data (2012–2024), we mapped age-standardized prevalence rates (ASPR) across the Czech Republic to explore the spatial distribution of PD. This unique nationwide cohort (representing all ≈48,000 diagnosed cases in the country), linked to population data, provides a detailed spatial and temporal overview of the situation in the Czech Republic and contributes to a still limited body of fine-scale research in Europe. Even after accounting for age, clear regional patterns remained, suggesting influences beyond demographics. These findings highlight the potential of spatial epidemiology to uncover signals of the environmental exposome. By capturing geographic variation, this approach opens new opportunities to investigate how environmental exposures, such as air pollution and pesticides, may contribute to PD risk.

Rational Engineering of Renilla reniformis Green Fluorescent Protein
Daniel Pluskal

Bioluminescence has wide applicability in science and medicine. The marine coral Renilla reniformis provides a highly efficient BRET-based bioluminescent system that has been previously adapted into the BRET-based probe IRrIS, though it is now limited to green emission. This work aimed to shift its spectral range by engineering its fluorescent protein (RGFP) toward longer wavelengths while maintaining its biological compatibility with its luciferase (RLuc). Targeted mutagenesis, guided by sequence–structure analysis, produced yellow-emitting variants (RYFP), with the best carrying mutations S66G and H197Y. However, these variants showed low fluorescence intensity, which was attributed to improper fluorophore charge and impaired maturation. These findings highlight key limitations but also provide a basis for further protein stabilisation and development of improved IRrIS probes.

Non-canonical amino acids in protein engineering: high efficiency click chemistry platform for protein immobilization
Diego Capelli

This study presents a platform for site-specific immobilization of FGF2-STAB® to overcome the limitations of random conjugation. Through in-silico design, internal methionines were replaced with leucine to isolate a single reactive site for azidohomoalanine (AHA) incorporation via selective pressure incorporation (SPI). Following rapid screening with Cell-Free Protein Synthesis (CFPS), variants were immobilized onto alkyne-functionalized agarose using copper-free SPAAC click chemistry. Results demonstrate a 95% coupling efficiency and full preservation of bioactivity in BaF3 2C cell assays. Current research is now focused on expanding this platform to include the site-specific clicking of fluorophores for advanced imaging applications.

Building a Human-Relevant Model to Uncover Pollutant Effects on Male Reproductive Health
Eliška Řehůřková

Environmental chemicals and pollutants can profoundly influence human reproductive health, yet their specific effects on male endocrine function remain insufficiently characterized. Progress in this area is hindered by the limitations of existing models used to study androgen synthesis in Leydig cells. To overcome these constraints, I am establishing an in vitro platform of Leydig‑like cells differentiated from human induced pluripotent stem cells (hiPSCs). By integrating 3D culture with a simulated microgravity, this system more closely reflects human physiological investigation of how chemicals influence Leydig cell development and function during sensitive stages. In addition to advancing mechanistic understanding, this human‑relevant model supports ethical research practices and contributes to improved chemical safety evaluation and male reproductive health studies.

Fully-Automated and Derivatization-free GC-MS Analysis of SCFA in Human Biospecimen Implementing Full Scan-SIM Data Acquisition
Hana Seličová

Short-chain fatty acids (SCFA), produced by gut microbiota, are essential for host metabolism and are linked to diseases including IBD, obesity, diabetes, and colorectal cancer, highlighting the need for scalable quantification methods. GC-MS is widely used but typically requires derivatization, increasing analysis time and toxicity, while recent direct methods still involve complex sample preparation. We developed a fully automated, derivatization-free GC-Q-MS method integrating acidification and MTBE extraction overlapped with data acquisition, reducing preanalytical error and achieving a 15-min sample cycle. The method enables quantification of nine SCFA (C2–C7, including branched isoforms). Addition of combined full-scan–SIM acquisition mode allows simultaneous targeted quantification and untargeted metabolite profiling. The standardized workflow supports reproducible, high-throughput clinical metabolomics across laboratories.

Impact of floods on soil quality and its safety
Lucie Jašová

Flood events strongly affect alluvial soils by mobilizing contaminated sediments and redistributing pollutants. Chemical analyses confirmed pre‑existing contamination, while the flood further modified concentrations and revealed contributions from local sources. Human‑risk predictions showed that some locations remain chronically contaminated, whereas others were impacted mainly by the flood. The RISKECO assessment indicates persistent ecological risk, although natural resilience helps maintain ecosystem functions. Ecotoxicological assays using two soil invertebrates, lettuce, and five microbial tests evaluated biological responses to the flood. E. crypticus survival decreased more after the flood, while reproduction was less affected. F. candida showed the opposite pattern. L. sativa responded more negatively to pre‑flood samples. Microbial activity was mostly stimulated, except for nitrification.

Dietary Bisphenol E Drives Expansion of Potentially Pathogenic Nocardia in Zebrafish Gut Microbiome
Michaela Vykypělová

Bisphenol E (BPE) is increasingly used as an alternative to restricted bisphenol A (BPA), although its biological effects remain poorly understood. This study investigated the impact of chronic dietary BPE exposure on the zebrafish gut microbiome. Adult zebrafish were exposed to BPE-enriched diet for 30 days. Gut microbiome composition was analyzed using 16S rRNA sequencing together with bioinformatical prediction of metabolic functional changes. BPE concentrations in muscle tissue were measured by LC-MS/MS to confirm uptake and bioaccumulation. While overall microbiome structure and diversity did not change, BPE exposure selectively enriched the potentially pathogenic genus Nocardia. Functional predictions further suggested microbiome responses associated with chemical stress. These findings indicate that BPE may not be a biologically neutral BPA alternative.

HDX-MS Pinpoints ApoE4 Structural Weakness Underlying Neurodegenerative Disease and Reveals Taurine/SPA as Selective Correctors
Naina Verma

ApoE4, the strongest genetic risk factor for late-onset Alzheimer’s disease, promotes earlier onset and faster neurodegeneration than ApoE3 through the destabilizing C112R mutation. Using HDX-MS with rapid online digestion and timsTOF SCP detection, we identified increased deuterium uptake in ApoE4 across helix 2, helix 3, the receptor-binding region, and areas adjacent to C112R, indicating pathogenic conformational opening absent in ApoE3. Molecular dynamics linked these regions to aggregation-prone ApoE4 dimers. Taurine and 3-sulfopropanoic acid selectively stabilized vulnerable regions, reduced conformational opening, suppressed pathogenic dimers, and decreased aggregation, while ApoE3 remained largely unaffected. In ApoE4/E4 cerebral organoids, taurine shifted cellular profiles toward an ApoE3-like state, supporting therapeutic potential against ApoE4-associated neurodegeneration. These findings demonstrate HDX-MS for defining structural vulnerabilities while monitoring correction.

Microbiome and Pharmacotherapy in Patients with Gastrointestinal Disorders - Research Highlights
Natálie Mlčůchová

Alterations in the gastrointestinal tract (GIT) microbiome are associated with genetic and exposure factors, such as drug use. The aims were to i) characterize the GIT bacteriome in patients with acid-peptic disease (APD, e.g., reflux) for whom proton pump inhibitors (e.g., lansoprazole) are indicated; ii) perform therapeutic monitoring of lansoprazole in ICU patients with APD risk; and iii) summarize variability in transporter genes that are key to the metformin metabolism; this antidiabetic/anticancer drug may influence the GIT microbiome. The GIT bacteriome profiling revealed complication/drug-related differences in patients with reflux. In ICU patients, non-genetic factors have a greater influence on lansoprazole metabolism than the patient’s genetic predisposition. A conceptual framework was proposed to improve the comparability and interpretation of pharmacogenetic studies.

Biomarkers of intertissue dialogue between muscle, adipose and bone tissue
Simona Holotová

Recent advances identify the Adipose–Muscle–Bone Axis as a sophisticated endocrine network regulated by biochemical crosstalk via adipokines, myokines, and osteokines. Dysregulation of this axis contributes to formation of different pathologies which significantly increasing discomfort and frailty risks not only in aging populations. This research examines these interactions through two clinical studies. Proteomic analysis of 32 bariatric patients identified 15 adipose-derived proteins correlating with DXA-derived bone parameters. Additionally, an intervention involving 103 older adults revealed that muscle-secreted follistatin-like proteins (FSTL1/FSTL5) significantly correlate with bone strength and density following exercise. These results demonstrate that bone health is biochemically modulated by adipose and muscle tissues beyond mechanical stress. Decoding this interconnected axis is vital for managing age-related metabolic and musculoskeletal decline.

The Urban Brain: Low-Level Visual Features Increase Metabolic Demand in the Visual Cortex
Tadeáš Dvořák

Merely viewing urban spaces can increase physiological and neural stress, yet the specific visual triggers remain unclear. In this study, 67 participants viewed urban and natural scenes during simultaneous fMRI, EEG, and eye-tracking. We applied computational algorithms to extract low-level image statistics, including luminance contrast and local chromatic differences. We found the human visual system reacts strongly to unnatural distributions of spatial features. Urban scenes with higher local chromatic differences evoked greater metabolic demand (in the early visual cortex) and increased saccadic exploration. Furthermore, higher luminance contrast was associated with reduced perceptual fluency, evidenced by decreased occipital alpha oscillations. These results suggest the unnatural visual statistics of cities impose a distinct metabolic load on the brain, contributing to urban stress.

Methods for the Assessment of Deiodinases Inhibition by Thyroid Hormone Disrupting Chemicals and Environmental Pollutant Mixtures Tereza Nešporová

Thyroid hormone (TH) regulation is vital for organism function, involving different organs across species. Disruption can cause developmental issues like decreased IQ, altered brain structure, attention deficits, mental retardation, or obesity. Iodothyronine deiodinases (DIOs) are key enzymes managing TH conversion. DIO imbalance is an important molecular initiating event (MIE) in the adverse outcome pathway (AOP) for thyroid disruption, affecting vertebrates, including fish and amphibians. Our study focuses on DIO1 and DIO3, developing human kidney HEK 293 cell lines with lentiviral overexpression of these enzymes. The assay detects iodine release from deiodination via the Sandell-Kolthoff reaction in microtiter plates.

Full-Size Structural and Energetic Modelling of the Staphylokinase Active Complex
Tim Titus Hendrikus de Martines

Ischaemic stroke remains a leading cause of death worldwide, yet treatment still relies on recombinant tissue plasminogen activators such as Alteplase and Tenecteplase, which have limited efficacy, high costs, and potentially severe side effects. Staphylokinase offers a promising alternative due to its high fibrin specificity, but its clinical use is hindered by immunogenicity and suboptimal activity. To address this, we have developed a filtering strategy for variant selection. Benchmarking identified FoldX as the most reliable optimisation tool. Subsequent saturation mutagenesis assessing stability and affinity can be used to guide development of improved staphylokinase variants. In addition, we are working on generating a full-size model for enhanced accuracy.

Acute vs. Chronic PFAS Exposure: Differential Thyroid Hormone Disruptions in Firefighter Cohorts
Soňa Skřídlová

Per- and polyfluoroalkyl substances (PFAS), often present in aqueous film-forming foams (AFFF), represent an occupational exposure concern for firefighters. We evaluated PFAS exposures and thyroid function in 165 males across three cohorts: newly recruited firefighters following acute AFFF training, career professionals, and non-firefighter controls. Serum concentrations of 7 PFAS and 5 thyroid hormones were analyzed using Generalized Linear Models and Quantile G-computation (QG). In novices, GLM showed PFUnDA and PFDA were significantly associated with elevated (F)T3. QG models further showed PFAS mixtures were linked to elevated (F)T3 and lower TSH. These specific associations were absent in professionals. This likely reflects a lack of recent high-dose exposure or a homeostatic adaptation to chronic burdens.