Fakultní nemocnice Bulovka

We present datasets from three large-scale human-subject experiments involving red-team hacking in a cyber range in the Guarding Against Malicious Biased Threats (GAMBiT) project. Across Experiments 1-3 (July 2024-March 2025), 19-20 skilled attackers per experiment conducted two 8-hour days of self-paced operations in a simulated enterprise network (SimSpace Cyber Force Platform) while collecting multi-modal data: self-reports (background, demographics, psychometrics), operational notes, terminal histories, key logs, network packet captures (PCAP), and NIDS alerts (Suricata). Each participant began from a standardized Kali Linux VM and pursued realistic objectives (e.g., target discovery and data exfiltration) under controlled constraints. Derivative curated logs and labels are included. The combined data release supports research on attacker behavior modeling, bias-aware analytics, and method benchmarking. Data are available via IEEE DataPort entries for Experiments 1-3.

IntroductionPrehabilitation is an increasingly recognised strategy designed to optimise patients’ physical, nutritional, and psychological status before major surgery. While the benefits of trimodal prehabilitation have been documented, we developed and implemented a quadrimodal model that incorporates occupational therapy as a fourth component. This comprehensive intervention targets frail patients with advanced oncogynecologic disease, a population at high risk for postoperative complications.Material and methodsThe study recruited 40 patients, 32 of whom successfully completed an intensive 3-week prehabilitation programme. The programme comprised four modules: physiotherapy, nutritional support, psychological support, and occupational therapy. Patients completed 4 days of inpatient care and 3 days of home care per week. The effectiveness of the programme was assessed using a range of methods, including physical tests (6-min walk test, 5 × sit-to-stand test), psychological status questionnaires (perceived stress scale) and nutritional assessment using established questionnaires (MUST and PONS).ResultsThe results demonstrated statistically significant enhancements in physical fitness assessments, a decline in the frailty index, and an augmentation in psychological well-being. Notably, three patients no longer fulfilled the frailty criteria at the conclusion of the programme. Spirometric parameters, including FVC, FEV1, and PEF, exhibited substantial improvement. The nutritional intervention resulted in a reduction in the number of patients at high risk of malnutrition. Additionally, there was a notable improvement in the nutritional parameters.ConclusionThis quadrimodal prehabilitation model, integrating occupational therapy, demonstrated effectiveness in preparing frail oncologic patients for radical surgery. The approach not only improved physical and psychological readiness but also allowed for patient stratification into responders and non-responders. Its implementation in clinical practice could enhance patient outcomes and reduce the burden of postoperative recovery.

Background/Objectives The leopard (Panthera pardus), an apex predator listed in CITES Appendix I and classified as Vulnerable by the IUCN, is undergoing severe population declines driven by habitat loss, human-wildlife conflict, and illegal trade. Rapid and reliable species and individual identification is critical for conservation and forensic applications, particularly when analyzing highly processed or degraded seized wildlife products, where morphological identification is often impossible. We aimed to develop and validate a robust multiplex quantitative real-time PCR (qPCR) assay combined with a short tandem repeat (STR) system for the species-specific detection and individual identification of P. pardus. Methods The qPCR assay (Ppar Qplex) was designed to target a mitochondrial Cytochrome b (Cyt b) fragment for species confirmation, a nuclear marker (PLP) for general Feliformia detection and quantification, and an artificial internal positive control (IPC) to monitor PCR inhibition. The assay's performance was validated for robustness, specificity, sensitivity, repeatability, and reproducibility, utilizing DNA extracted from 30 P. pardus individuals (hair and feces) and tested against 18 related Feliformia species and two outgroups. Individual identification was achieved using a set of 18 STR loci and a sex determination system adapted from previously published Panthera panels. Results Validation demonstrated high specificity for the Ppar Qplex: mitochondrial amplification occurred exclusively in P. pardus samples. The nuclear marker consistently amplified across all 18 tested Feliformia species but not the outgroups. The assay showed high analytical sensitivity, successfully detecting DNA at concentrations as low as 1 pg/µL, with consistent results confirmed across different sample types, replicates, and independent users. Furthermore, the STR multiplex successfully generated 30 unique individual profiles using the 18 polymorphic loci and the sex determination system. Conclusions The combined qPCR assay and STR system provide a fast, sensitive, and highly specific molecular framework for rapid leopard detection, quantification, and individual identification from a wide range of sample types. These tools strengthen forensic capacity to combat wildlife crime and provide critical data to support evidence-based conservation management of P. pardus. P. pardus, an apex predator listed in CITES Appendix I and classified as Vulnerable by the IUCN, is undergoing severe population declines driven by habitat loss, human-wildlife conflict, and illegal trade. Rapid and reliable identification of seized specimens is therefore critical for conservation and forensic applications, mainly when products are highly processed. We developed and validated a multiplex quantitative real-time PCR (qPCR) assay targeting the mitochondrial gene Cytochrome b (Cyt b) for species-specific detection. The assay was tested on verified leopard individuals and validated across 18 Feliformia and two outgroup species (Homo sapiens, Canis lupus familiaris). Analytical performance was assessed through robustness, specificity, sensitivity, repeatability, and reproducibility. Mitochondrial amplification occurred exclusively in leopard samples, while nuclear markers amplified consistently across Feliformia but not in outgroup species. The assay's limit of DNA detection is 1 pg/µL and produces consistent results across replicates, tested types of samples (hair, feces), and independent users, with internal controls confirming the absence of inhibition. In addition, we present the results of successful individual identification using the set of 18 STR loci and the sex determination system. The developed qPCR and STR systems provide a fast, sensitive, and specific solution for leopard detection and quantification, reinforcing forensic efforts against wildlife crime and supporting conservation of P. pardus.

ABSTRACTAimThis study aimed to investigate the incidence and severity of acute poststreptococcal glomerulonephritis (APSGN) in the Czech paediatric population before, during and after the coronavirus disease 2019 (COVID‐19) pandemic.MethodsThe data were collected from 15 centres. The patients were divided into three periods: 2016–2019 (pre‐COVID), 2020–2021 (COVID) and 2022–2023 (post‐COVID). The incidence of APSGN during the studied period was assessed. The clinical and laboratory parameters were compared between the pre‐COVID and post‐COVID periods.ResultsOf the 117 patients with possible APSGN, 95 met the inclusion criteria. There were 67 (70.5%) males. The incidence of APSGN was significantly higher during the post‐COVID period (32/year) than during the pre‐COVID (6/year) and COVID (3.5/year) periods (p < 0.0001). The highest number of cases occurred in 2023 (n = 55). During the post‐COVID period there was more frequent use (p = 0.0448) and duration (p = 0.0131) of furosemide; more frequent antihypertensive treatment (p = 0.0104); and a higher prevalence of acute kidney injury (p = 0.0145), hypertension (p = 0.0178), and hypertension stage 2 (p = 0.0372) compared to the pre‐COVID period. Kidney replacement therapy and crescentic glomerulonephritis were found only during the post‐COVID period.ConclusionWe found a significantly higher incidence of APSGN and its associated complications during the post‐COVID period.

Abstract Background Dupuytren’s disease (DD) is a common fibrotic disorder of the hand, characterized by progressive thickening and contracture of the palmar and digital fascia. Surgical excision remains the primary treatment; however, there are currently no therapies to prevent disease progression or recurrence. This study aims to develop a 3D in vitro model to test novel antifibrotic therapies. The model is based on decellularized pathological DD tissue seeded with patient-derived fibroblasts, capturing the role of both cellular and extracellular matrix components in disease progression. Methods Fibrotic DD tissues were obtained from surgical excisions, sectioned, and decellularized. In parallel, primary fibroblasts were isolated from patient samples. The decellularized extracellular matrices (dECMs) were characterized with respect to biochemical composition, collagen structure, and mechanical properties. Fibroblasts were seeded onto the dECMs and cultured stepwise to initially promote proliferation, followed by differentiation into myofibroblasts. Secretomes of cells cultivated on the established 3D model were compared to those from conventional 2D cultivations. To evaluate the model´s relevance and effectiveness we tested the antifibrotic drug minoxidil. Results The dECMs retained the pathological architecture and mechanical properties of native DD tissue, although individual ECM components were reduced after decellularization. Fibroblasts successfully adhered, proliferated, and repopulated the scaffold. The relevance of the 3D model was demonstrated by the presence of myofibroblasts with disease–relevant secretome. The responsiveness to the drug minoxidil was significantly more complex in the 3D model than in conventional 2D cultures. Conclusion We demonstrated that dECM seeded with DD fibroblasts represents a relevant 3D in vitro model of Dupuytren’s disease. The model enables antifibrotic drug screening, as demonstrated by the testing of minoxidil. Our model provides a reproducible platform also suitable for the investigation of cells and ECM contributions to fibrotic processes.

Modern tissue engineering requires not only degradable materials promoting cell growth and differentiation, but also vascularization of the engineered tissue. Porous polylactide/polycaprolactone (PLA/PCL, ratio 3/5) foam scaffolds were prepared by a combined porogen leaching and freeze-drying technique using NaCl (crystal size 250-500 µm) and a water-soluble cellulose derivative (KlucelTM E; 10-100% w/w relative to the total PLA/PCL concentration) as porogens. Scanning electron microscopy, micro-CT, and Brunauer-Emmett-Teller analysis showed that all scaffolds contained a trimodal range of pore sizes, i.e., macropores (average diameter 298-539 μm), micropores (100 nm to 10 μm), and nanopores (mostly around 3.0 nm). All scaffolds had an open porosity of about 90%, and the pores were interconnected. The size of the macropores and the nanoporosity were higher in the scaffolds prepared with Klucel. Nanoporosity increased water uptake by the scaffolds, while macroporosity promoted cell ingrowth, which was most evident in scaffolds prepared with 25% Klucel. Human adipose-derived stem cells co-cultured with endothelial cells formed pre-vascular structures in the scaffolds, which was further enhanced in a dynamic cell culture system. The scaffolds are promising for the engineering of pre-vascularized soft tissues (relatively pliable 10% Klucel scaffolds) and hard tissues (mechanically stronger 25% and 50% Klucel scaffolds).

Primary tumors of the calcaneus present a reconstructive challenge. Their relatively low incidence leads to limited evidence, clinical experience, and ongoing discussion regarding the optimal surgical strategy. In this report, we present a case of a 19-year-old male diagnosed with an aggressive osteoblastoma who underwent subtotal calcanectomy. A custom 3D-printed titanium endoprosthesis was designed for limb-salvage reconstruction. Trabecular porous structure and screws were used to achieve subtalar and calcaneo-cuboid fusion and to compensate for the deficient ligamentous apparatus following resection. An innovative system for reattachment of the calcaneal tuberosity with spared of the Achilles tendon was also utilized. At 3 months, the implant was completely incorporated, allowing full weight bearing. At the 18-month follow-up, the implant showed no signs of loosening, the patient had no signs of recurrence, had excellent function, and successfully returned to all his pretreatment activities, underscoring the effectiveness of this reconstructive approach in the management of calcaneal tumors.Level of Evidence: IV, Case report.

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