UNLABELLED: The aim of this study was to identify parameters influencing DNA extraction and PCR amplification efficiencies in an attempt to standardize Mucorales qPCR. The Fungal PCR Initiative Mucorales Laboratory Working Group distributed two panels of simulated samples to 26 laboratories: Panel A (six sera spiked with Mucorales DNA and one negative control serum) and Panel B (six Mucorales DNA extracts). Panel A underwent DNA extraction in each laboratory according to the local procedure and were sent to a central laboratory for testing using three different qPCR techniques: one in-house qPCR assay and two commercial assays (MucorGenius and Fungiplex). Panel B DNA extracts were PCR amplified in each laboratory using local procedures: nine in-house qPCR assays and two commercial kits (MucorGenius and MycoGENIE). All data were compiled and anonymously analyzed at the central laboratory. For Panel A, a total of six different automated platforms and five manual extraction methods were used. Positive rates were 64%, 70%, and 89%, for the MucorGenius, Fungiplex, and the in-house qPCR assay, respectively. Using a large volume of serum for DNA extraction provided the highest analytical sensitivity (82.5% for 1 mL compared with 62.7% for smaller volumes, P < 0.01). For Panel B, five in-house qPCR assays and two commercial kits had >78% positivity. Using larger PCR input volumes (≥7 μL) was associated with the highest sensitivity at 95.5% compared to 58.3% when lower input volumes were used (P < 0.01). Using larger sample volumes for nucleic acid extraction and DNA template volumes for PCR amplification significantly improves the performance of Mucorales qPCR when testing serum. IMPORTANCE: Mucormycosis is a life-threatening mold infection affecting immunosuppressed patients but also other patients with diabetes or trauma. Better survival is linked to shorter delays in diagnosis and treatment initiation. Detection of Mucorales-free DNA in serum or plasma using quantitative PCR allows a prompt diagnosis and earlier treatment. Several techniques and protocols of quantitative Mucorales PCR are used in Europe, and improving performance remains a common objective of laboratories participating in the fungal PCR Initiative Working Group. This study, which combined results from 26 laboratories in Europe, showed that the main parameters underpinning sensitivity are the preanalytical variables (volume of serum used for DNA extraction and DNA template volume), irrespective of the extraction platforms and qPCR assay/platform.

• MeSH
• diagnostické techniky molekulární normy   metody   MeSH
• DNA fungální * krev   genetika   MeSH
• kvantitativní polymerázová řetězová reakce * normy   metody   MeSH
• lidé MeSH
• Mucorales * genetika   izolace a purifikace   MeSH
• mukormykóza * diagnóza   mikrobiologie   krev   MeSH
• senzitivita a specificita * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH

BACKGROUND: The increasing complexity of mass casualty incidents (MCIs) necessitates highly effective training for emergency responders. Traditional training methods, while effective in teaching core skills, often fail to replicate the dynamic, high-pressure environments responders face in real-world crises. Virtual reality (VR) offers a novel approach to emergency training, providing an immersive, controlled setting that can simulate real-life scenarios. This study explores the effectiveness of VR in training paramedic students for MCIs and compares the outcomes to those from conventional training methods. METHODS: A comparative study was conducted with 37 paramedic students who underwent either VR-based training or conventional training using mannequins and real-world equipment. The VR application simulated a mass casualty car accident, focusing on triage and patient management. Both groups were assessed based on their performance in key areas, including the accuracy of situational reporting (METHANE), patient triage, heart rate monitoring, and perceived demand using the NASA Task Load Index (NASA-TLX). RESULTS: The VR group demonstrated significantly lower mental demand (p < 0.001) and frustration levels (p = 0.021) compared to traditional training. However, task completion times were slower in the VR setting (p < 0.001), likely due to the interface's unfamiliarity. Accuracy in situational reporting was higher in VR (p = 0.002), while heart rate monitoring did not reveal a significant difference between the groups (p = 0.516). Although VR did not reduce temporal demand (p = 0.057), it showed potential for improving focus and precision in training. Error rates in triage were similar across both training methods (p = 0.882), indicating comparable performance levels in patient classification. CONCLUSIONS: VR presents a promising tool for training emergency responders, particularly in situations that require rapid upskilling, such as crises or wars. The ability to simulate realistic, high-pressure scenarios in a controlled environment can enhance both cognitive and emotional preparedness. Further research is necessary to optimize VR systems and interfaces, making them more efficient for real-time decision-making. As VR technology advances, it holds potential as a key component in future emergency preparedness strategies.

• MeSH
• dospělí MeSH
• first responder * výchova   psychologie   MeSH
• hromadné neštěstí * MeSH
• lidé MeSH
• plánování postupu v případě katastrof metody   MeSH
• třídění pacientů metody   MeSH
• virtuální realita * MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVES: Regular physical activity (PA) and reduced sedentary behaviour (SB) have been associated with positive health outcomes, but many older adults do not comply with the current recommendations. Sensor-triggered ecological momentary assessment (EMA) studies allow capturing real-time data during or immediately after PA or SB, which can yield important insights into these behaviours. Despite the promising potential of sensor-triggered EMA, this methodology is still in its infancy. Addressing methodological challenges in sensor-triggered EMA studies is essential for improving protocol adherence and enhancing validity. Therefore, this study aimed to examine (1) the patterns in sensor-triggered EMA protocol adherence (eg, compliance rates), (2) the impact of specific settings (eg, event duration) on the number of prompted surveys, and (3) participants' experiences with engaging in a sensor-triggered EMA study. DESIGN: Two longitudinal, sensor-triggered EMA studies-one focused on PA and the other on SB-were conducted using similar methodologies from February to October 2022. Participants' steps were monitored for seven days using a Fitbit activity tracker, which automatically prompted an EMA survey through the HealthReact smartphone application when specified (in)activity thresholds were reached. After the monitoring period, qualitative interviews were conducted. Data from both studies were merged. SETTING: The studies were conducted among community-dwelling Belgian older adults. PARTICIPANTS: The participants had a median age of 72 years, with 54.17% being females. The PA study included 88 participants (four dropped out), while the SB study included 76 participants (seven dropped out). PRIMARY AND SECONDARY OUTCOME MEASURES: Descriptive methods and generalised logistic mixed models were employed to analyse EMA adherence patterns. Simulations were conducted to assess the impact of particular settings on the number of prompted EMA surveys. Additionally, qualitative interview data were transcribed verbatim and thematically analysed using NVivo. RESULTS: Participants responded to 81.22% and 79.10% of the EMA surveys in the PA and SB study, respectively. The confirmation rate, defined as the percentage of EMA surveys in which participants confirmed the detected behaviour, was 94.16% for PA and 72.40% for SB. Logistic mixed models revealed that with each additional day in the study, the odds of responding to the EMA survey increased significantly by 1.59 times (OR=1.59, 95% CI: 1.36 to 1.86, p<0.01) in the SB study. This effect was not observed in the PA study. Furthermore, time in the study did not significantly impact the odds of participants confirming to be sedentary (OR=0.97, 95% CI: 0.92 to 1.02, p=0.28). However, it significantly influenced the odds of confirming PA (OR: 0.81, 95% CI: 0.68 to 0.97, p=0.02), with the likelihood of confirming decreasing by 19% with each additional day in the study. Furthermore, a one-minute increase in latency (ie, time between last syncing and starting the EMA survey) in the PA study decreased the odds of the participant confirming to be physically active by 20% (OR: 0.80, 95% CI: 0.72 to 0.89, p<0.01). Simulations of the specific EMA settings revealed that reducing the event duration and shorter minimum time intervals between prompts increased the number of EMA surveys. Overall, most participants found smartphone usage to be feasible and rated the HealthReact app as user-friendly. However, some reported issues, such as not hearing the notification, receiving prompts at an inappropriate time and encountering technical issues. While the majority reported that their behaviour remained unchanged due to study participation, some noted an increased awareness of their habits and felt more motivated to engage in PA. CONCLUSIONS: This study demonstrates the potential of sensor-triggered EMA to capture real-time data on PA and SB among older adults, showing strong adherence potential with compliance rates of approximately 80%. The SB study had lower confirmation rates than the PA study, due to technical issues and discrepancies between self-perception and device-based measurements. Practical recommendations were provided for future studies, including improvements in survey timing, technical reliability and strategies to reduce latency.

• MeSH
• cvičení * MeSH
• fitness náramky MeSH
• lidé MeSH
• longitudinální studie MeSH
• okamžité posouzení v přirozeném prostředí * MeSH
• samostatný způsob života * MeSH
• sedavý životní styl * MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Belgie MeSH

AIM: This study aimed to investigate the phytochemical composition of Psychotria montana extract (PME) and evaluate its inhibitory effects on MCF7 breast cancer cells. METHODS: The chemical composition of PME was analyzed using UPLC-QToF-MS. The effects of PME on cell proliferation were evaluated using the MTT assay. Flow cytometry was used for cell cycle and apoptosis analysis. The effects of PME on the transcription of cell cycle control genes were assessed using real-time PCR. RESULTS: UPLC-QToF-MS analysis revealed major compounds of PME, including terpenoids and flavonoids, with the potential to inhibit proliferation, migration, and induce apoptosis in MCF7 cancer cells. PME effectively suppressed MCF7 cell proliferation under 2D culture, with a low IC50 value of 34.7 μg/ml. PME also hindered cell migration (p < 0.01) and reduced spheroid number (p < 0.001) and size (p < 0.001) in serum-free 3D culture. Apoptosis analysis via nuclear staining with DAPI and flow cytometry revealed an increase in the number of apoptotic cells after PME treatment (p < 0.001). Additionally, the PME induced cell cycle arrest at the G0/G1 phase (p < 0.05). PME altered the expression of cell cycle control genes (cyclins and CDKs) as well as cancer suppressor genes including p16, p27, and p53 at the transcriptional level (mRNA). The results of molecular docking suggest that the compounds present in PME exhibit a high binding affinity for CDK3, CDK4, CDK6, and CDK8 proteins, which are essential regulators of the cell cycle. CONCLUSION: Psychotria montana has the potential to inhibit cancer cells by inducing apoptosis and halting the cell cycle of MCF7 breast cancer cells.

• MeSH
• apoptóza * účinky léků   MeSH
• buněčný cyklus účinky léků   MeSH
• fytogenní protinádorové látky farmakologie   chemie   MeSH
• lidé MeSH
• MFC-7 buňky MeSH
• nádory prsu * farmakoterapie   patologie   genetika   metabolismus   MeSH
• počítačová simulace MeSH
• pohyb buněk účinky léků   MeSH
• proliferace buněk * účinky léků   MeSH
• Psychotria * chemie   MeSH
• rostlinné extrakty * farmakologie   chemie   MeSH
• simulace molekulového dockingu MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: The management of patients with non-dialysis dependent chronic kidney disease (NDD-CKD) is challenging due to coexisting diseases, competing risks and uncertainties around optimal transition planning. Such clinical challenges are further exacerbated by physician shortage, coupled with rising service demands, which may hinder timely medical access due to long waiting times. Accurate progression risk assessment may help optimize resource allocation and adapting care based on individual patients' needs. This study validated the Prognostic Reasoning System for Chronic Kidney Disease Progression (PROGRES-CKD) in an Italian public hospital and compared its potential impact on waiting list optimization against physician-based protocols. METHODS: First we first validated PROGRES-CKD by assessing its accuracy in predicting kidney replacement therapy (KRT) initiation within 6 months and 24 months in a historical cohort of patients treated at the San Gerardo Hospital (Italy) between 01-01-2015 and 31-12-2019. In a second study we compared PROGRES-CKD to attending nephrologists' prognostic ratings and simulated their potential impact on a waiting list management protocol. RESULTS: We included 2005 patients who underwent 11,757 outpatient nephrology visits in 4 years. Most visits occurred for NDD-CKD stage 4 patients; the incidence of KRT onset was 10.8 and 9.32/100 patient-years at the 6 and 24-month prediction horizon cohorts, respectively. PROGRES-CKD demonstrated high accuracy in predicting KRT initiation at 6 and 24 months (AUROC = 0.88 and AUROC = 0.85, respectively). Nephrologists' prognostic performance was highly operator-dependent, albeit always significantly lower than PROGRES-CKD. In the simulation exercise, allocation based on PROGRES-CKD resulted in more follow-up visits for patients progressing to end-stage kidney disease (ESKD) and fewer visits for non-progressing patients, compared to allocation determined by nephrologists' prognosis. CONCLUSIONS: PROGRES-CKD showed high accuracy in a real-world application. Waiting list simulation suggests that PROGRES-CKD may enable more efficient allocation of resources.

• MeSH
• ambulantní péče * MeSH
• časové faktory MeSH
• chronická renální insuficience * terapie   diagnóza   MeSH
• hodnocení rizik metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• náhrada funkce ledvin statistika a číselné údaje   MeSH
• nefrologie * MeSH
• nemocnice veřejné MeSH
• prognóza MeSH
• progrese nemoci MeSH
• senioři MeSH
• seznamy čekatelů * MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• validační studie MeSH
• Geografické názvy
• Itálie MeSH

A good knowledge of the theoretical foundations of medicine helps students and physicians to better recognize and treat patients with complex medical conditions, including sepsis and septic shock. The article describes the authors' experience in implementing the analysis of sepsis and septic shock using a high-fidelity simulated clinical scenario in the course of pathological physiology for preclinical medical students. The unique aspect of our approach is the integration of core physiology concepts, such as homeostasis, causality, structure-function relationships, and fundamental pathophysiology concepts (e.g., etiology, pathogenesis, cell and tissue damage, inflammation, symptoms, and syndromes) in the analysis of the patient's condition on the high-fidelity simulator with preclinical medical students. According to the students' feedback, the use of a high-fidelity simulator to analyze the sepsis and septic shock scenario increased their interest in the class, improved their motivation to learn the material, and helped them adapt in a safe environment to making decisions based on a large amount of data about a complex patient condition in a time-sensitive situation.NEW & NOTEWORTHY The authors applied core theoretical concepts of physiology and the fundamental concepts of pathological physiology for teaching sepsis and septic shock clinical scenarios on the high-fidelity simulator in the course of pathological physiology for preclinical medical students. It elevated students' interest and motivation, enhanced the educational experience, and prepared students better for real-world clinical decision-making. We consider that this idea might be an inspiration to colleagues and invite further discussion.

• MeSH
• fyziologie výchova   MeSH
• lidé MeSH
• sepse * patofyziologie   terapie   MeSH
• studenti lékařství * MeSH
• studium lékařství pregraduální metody   MeSH
• vyučování MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Arterial compliance (AC) is an important cardiovascular parameter characterizing mechanical properties of arteries. AC is significantly influenced by arterial wall structure and vasomotion, and it markedly influences cardiac load. A new method, based on a two-element Windkessel model, has been recently proposed for estimating AC as the ratio of the time constant T of the diastolic blood pressure decay and peripheral vascular resistance derived from clinically available stroke volume measurements and selected peripheral blood pressure parameters which are less prone to peripheral distortions. The aim of this study was to validate AC estimation using a virtual population generated by in silico model of the systemic arterial tree. In the second part of study, we analysed causal coupling between AC oscillations and variability of its potential determinants - systolic blood pressure and heart rate in healthy young human subjects. The pool of virtual subjects (n=3818) represented an extensive AC distribution. AC was estimated from the peripheral blood pressure curve and by the standard method from the aortic blood pressure curve. The proposed method slightly overestimated AC set in the model but both ACs were strongly correlated (r=0.94, p<0.001). In real data, we observed that AC dynamics was coupled with basic cardiovascular parameters variability independently of the autonomic nervous system state. In silico analysis suggests that AC can be reliably estimated by noninvasive method. The analysis of short-term AC variability together with its determinants could improve our understanding of factors involved in AC dynamics potentially improving assessment of AC changes associated with atherosclerosis process. Key words Arterial compliance, Cardiovascular model, Arterial blood pressure, Causal analysis, Volume-clamp photoplethysmography.

• MeSH
• arterie * fyziologie   MeSH
• cévní rezistence fyziologie   MeSH
• dospělí MeSH
• krevní tlak * fyziologie   MeSH
• lidé MeSH
• mladý dospělý MeSH
• modely kardiovaskulární * MeSH
• počítačová simulace * MeSH
• poddajnost MeSH
• srdeční frekvence fyziologie   MeSH
• tuhost cévní stěny fyziologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• validační studie MeSH

The mechanism of rotator cuff injury remains to be elucidated. And COX-2 plays a dual role in skeletal muscle injury and regeneration, would be associated with the development of rotator cuff injury. Therefore, we chose human skeletal muscle cells (HSKMC) as an in vitro muscle tissue model and transfected lentivirus with overexpressed COX-2 to simulate the in vitro environment of rotator cuff injury. To investigate the specific molecular biological mechanism of COX-2, transcriptome sequencing (RNA-Seq) was used to analyze the differentially expressed mRNAs in HSKMC overexpressing COX-2. Enrichment analysis was performed to analyze these differentially expressed genes and real-time quantitative PCR (RT-qPCR) was used to examine the mRNA levels of genes induced by overexpression. Subsequently, the role of COX-2 in cell proliferation was confirmed by cell counting kit-8 (CCK-8), and focal adhesion kinase (FAK) and signal transducer and activator of transcription 3 (STAT3) phosphorylation induced by COX-2 was utilized by western blotting (WB). The results showed that total of 30,759 differentially expressed genes were obtained, and the expression of CYP4F3 and GPR87 was significantly increased. COX-2 could bind CYP4F3 and GPR87 and co-localize with them in the cytoplasm. Finally, COX-2 promoted the proliferation of human skeletal muscle cells by activating the FAK and STAT3 pathways.

• MeSH
• cyklooxygenasa 2 * metabolismus   genetika   MeSH
• kosterní svalová vlákna metabolismus   enzymologie   patologie   MeSH
• kosterní svaly metabolismus   patologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• poranění rotátorové manžety * metabolismus   patologie   enzymologie   genetika   MeSH
• proliferace buněk MeSH
• transkripční faktor STAT3 metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

In this dynamic scanning protocol, ultrasound examination of the ankle is described using various maneuvers to assess different conditions. Real-time patient examination and scanning videos are used for better simulation of daily clinical practice. The protocol is prepared by several/international experts in the field of musculoskeletal ultrasound and within the umbrella of European Musculoskeletal Ultrasound Study Group in Physical and Rehabilitation Medicine/Ultrasound Study Group of the International Society of Physical and Rehabilitation Medicine.

• MeSH
• kotník diagnostické zobrazování   MeSH
• lidé MeSH
• muskuloskeletální nemoci * diagnostické zobrazování   rehabilitace   MeSH
• muskuloskeletální systém * diagnostické zobrazování   MeSH
• noha (od hlezna dolů) diagnostické zobrazování   MeSH
• rehabilitační lékařství * MeSH
• ultrasonografie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The purpose of this study was to verify the influence of contextual perception in sports on performance in reaction agility to a specific and non-specific stimulus on a sample of handball players. The object of the research was eleven players of the junior handball team playing in the top league (age: 16.73 years; decimal age 16.46 years). Visual signaling and video projection. and a modified Y-test of reaction agility were used. In this study. two goals were set: a) to compare the performance level in the Y agility test with respect to a simple non-specific and a complex specific visual stimulus in the form of filmed scenes of the opponent's attacking actions. which simulated real situations in performance handball; b) find out the possible relationship between performance in the agility test and reaction time. decision-making competence. anxiety and overload and involvement in decision-making and performance with regard to the nature of the stimulus. A questionnaire (CETD) was implemented in the sample to assess the decision-making style of athletes in a specific area.