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
- Molecular Diagnostic Techniques standards methods MeSH
- DNA, Fungal * blood genetics MeSH
- Real-Time Polymerase Chain Reaction * standards methods MeSH
- Humans MeSH
- Mucorales * genetics isolation & purification MeSH
- Mucormycosis * diagnosis microbiology blood MeSH
- Sensitivity and Specificity * MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
OBJECTIVES: Nonpharmacologic interventions (NPIs) constitute an important part of treatment for older adults, cover a broad and diverse range of interventions, and have advantages over pharmacologic interventions (eg, limited adverse side effects). However, an unambiguous definition of NPIs is still lacking. Defining NPIs may facilitate research on this topic and enhance comparability of results between studies, and might help to face the challenges of recognition, acceptation, funding, and implementation. Therefore, the aim of this review was to provide an overview and comparison of the definitions of NPIs used in the current literature on older adults. DESIGN: A systematic review was performed to provide an overview of the definitions of NPIs that are used in the current literature on older populations and to organize the characteristics involved in the definitions. SETTING AND PARTICIPANTS: People ≥60 years of age were included, not limited to a specific setting. METHODS: A systematic search was performed in the following 5 databases: PubMed, Embase, Clarivate Analytics/Web of Science Core Collection, Cumulative Index to Nursing and Allied Health Literature, and Wiley/Cochrane Library. The time frame within the databases was from inception to December 4, 2023. Review articles, editorials and consensus papers were included. RESULTS: We included 28 articles. We organized the definitions of NPI according to 4 different aspects: types of interventions involved, target population, goals the interventions addressed, and requirements of the interventions. Definitions in the current literature can generally be divided into 2 groups: NPIs described as not involving medication, and more elaborated multidomain definitions. Based on the results, we formulated criteria for types of interventions that can be considered an NPI. CONCLUSIONS AND IMPLICATIONS: Using current descriptions and characteristics, elements for a new definition for NPIs were proposed. To improve research in this field, consensus needs to be reached regarding elements covered by a definition of NPIs.
- MeSH
- Middle Aged MeSH
- Humans MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Therapeutics * methods MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Systematic Review MeSH
OBJECTIVE: Preeclampsia is a major pregnancy complication that results in significant maternal and infant mortality and morbidity, yet difficulties remain in the diagnosis of preeclampsia based on clinical parameters alone. The objective was to assess the performance of a hand-held point-of-care (POC) immunoassay in a clinical environment for glycosylated fibronectin (GlyFn) for the prediction of preeclampsia within 4 weeks of sampling. METHODS: Multinational European prospective observational pilot study of predominantly high-risk patients in the second half of pregnancy to assess a point-of-care immunoassay for GlyFn in predicting preeclampsia within 4 weeks of sampling. GlyFn was measured using a second generation hand held POC immunoassay. Results were considered normal for GlyFn concentrations of < 350 μg/mL, positive for GlyFn concentrations of 351-600 μg/mL, and high-positive for GlyFn concentrations > 600 μg/mL. RESULTS: Preeclampsia developed in 16 (19%) of 84 subjects and was associated with a shorter gestational age at delivery 35.3 weeks vs. 37.3 weeks for non-preeclamptics, n = 82; p = 0.001), a higher risk of fetal growth restriction (FGR; 31.2% vs. 10.3% for non-preeclamptics, p = 0.046), and an increased risk of preterm birth < 37 weeks gestation (83.3% vs. 33.3% for non-preeclamptics, (n = 78; p = 0.003). GlyFn positive or high positive was seen in 13/16 (81%) and in 35/68 (51.5%), yielding a sensitivity of 81%, a specificity of 49%, a positive predictive value of 27%, and a negative predictive value of 92%. GlyFn positive or high positive was also associated with preterm birth < 37 weeks in singleton pregnancy non-preeclamptic patients. Preterm birth occurred in 4.8% of those with normal GlyFn, in 26.7% with positive GlyFn, and in 50% of those with high GlyFn in singleton gestations without preeclampsia (p = 0.008). CONCLUSION: The ability to use this test in a POC format provides a method for practitioners to quickly determine risk for preeclampsia in their pregnant patients and offers an affordable alternative, as a single analyte to other diagnostic or screening tests that require laboratory-based testing or ultrasound equipment. Independent of preeclampsia, an elevated GlyFn was also correlated with preterm delivery and requires further study.
- MeSH
- Biomarkers analysis MeSH
- Adult MeSH
- Fibronectins * analysis MeSH
- Gestational Age MeSH
- Glycated Proteins MeSH
- Immunoassay methods MeSH
- Humans MeSH
- Pilot Projects MeSH
- Point-of-Care Testing MeSH
- Predictive Value of Tests MeSH
- Pre-Eclampsia * diagnosis MeSH
- Prospective Studies MeSH
- Pregnancy MeSH
- Point-of-Care Systems MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Pregnancy MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Multicenter Study MeSH
- Observational Study MeSH
OBJECTIVES: Illicit drug use presents a significant challenge to global health and public safety, requiring innovative and effective monitoring strategies. This study aimed to evaluate the current landscape of wastewater-based epidemiology (WBE) for monitoring illicit drugs in Europe, focusing on collaboration, current practices, and barriers, while identifying opportunities for improvement. STUDY DESIGN: Cross-sectional survey-based study. METHODS: Coordinated by the Sewage Analysis CORe Group Europe (SCORE) and the European Union Drugs Agency (EUDA), two surveys were conducted in 2023 targeting researchers and stakeholders using WBE for illicit drugs. Data were analysed to identify trends, gaps, and opportunities for improving WBE implementation. RESULTS: The findings indicate a robust research infrastructure and diverse analytical methods among European institutions. Two-thirds of the participating countries reported using WBE data to inform policy. However, challenges persist, particularly in securing funding and coordination, as well as generating national estimates from multiple locations and addressing specific local policy needs. CONCLUSIONS: WBE has proven to be a valuable tool for monitoring illicit drug trends and informing drug policies. To unlock its full potential, sustained funding, methodological standardization, and enhanced cooperation are essential. This study provides critical insights into the European WBE landscape, offering a roadmap for strengthening the integration of actionable WBE data into public health and policy frameworks.
- MeSH
- Wastewater-Based Epidemiological Monitoring MeSH
- Humans MeSH
- Wastewater * MeSH
- Substance-Related Disorders epidemiology MeSH
- Cross-Sectional Studies MeSH
- Surveys and Questionnaires MeSH
- Decision Making MeSH
- Illicit Drugs * MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Europe MeSH
BACKGROUND: Observational data on composite scores often comes with missing component information. When a complete-case (CC) analysis of composite scores is unbiased, preferable approaches of dealing with missing component information should also be unbiased and provide a more precise estimate. We assessed the performance of several methods compared to CC analysis in estimating the means of common composite scores used in axial spondyloarthritis research. METHODS: Individual mean imputation (IMI), the modified formula method (MF), overall mean imputation (OMI), and multiple imputation of missing component values (MI) were assessed either analytically or by means of simulations from available data collected across Europe. Their performance in estimating the means of the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), the Bath Ankylosing Spondylitis Functional Index (BASFI), and the Ankylosing Spondylitis Disease Activity Score based on C-reactive protein (ASDAS-CRP) in cases where component information was set missing completely at random was compared to the CC approach based on bias, variance, and coverage. RESULTS: Like the MF method, IMI uses a modified formula for observations with missing components resulting in modified composite scores. In the case of an unbiased CC approach, these two methods yielded representative samples of the distribution arising from a mixture of the original and modified composite scores, which, however, could not be considered the same as the distribution of the original score. The IMI and MF method are, thus, intrinsically biased. OMI provided an unbiased mean but displayed a complex dependence structure among observations that, if not accounted for, resulted in severe coverage issues. MI improved precision compared to CC and gave unbiased means and proper coverage as long as the extent of missingness was not too large. CONCLUSIONS: MI of missing component values was the only method found successful in retaining CC's unbiasedness and in providing increased precision for estimating the means of BASDAI, BASFI, and ASDAS-CRP. However, since MI is susceptible to incorrect implementation and its performance may become questionable with increasing missingness, we consider the implementation of an error-free CC approach a valid and valuable option. TRIAL REGISTRATION: Not applicable as study uses data from patient registries.
- MeSH
- Spondylitis, Ankylosing MeSH
- Axial Spondyloarthritis * MeSH
- C-Reactive Protein analysis MeSH
- Data Interpretation, Statistical MeSH
- Humans MeSH
- Severity of Illness Index MeSH
- Research Design MeSH
- Bias MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Europe MeSH
We present new developments for an ab-initio model of the neutron relative biological effectiveness (RBE) in inducing specific classes of DNA damage. RBE is evaluated as a function of the incident neutron energy and of the depth inside a human-sized reference spherical phantom. The adopted mechanistic approach traces neutron RBE back to its origin, i.e. neutron physical interactions with biological tissues. To this aim, we combined the simulation of radiation transport through biological matter, performed with the Monte Carlo code PHITS, and the prediction of DNA damage using analytical formulas, which ground on a large database of biophysical radiation track structure simulations performed with the code PARTRAC. In particular, two classes of DNA damage were considered: sites and clusters of double-strand breaks (DSBs), which are known to be correlated with cell fate following radiation exposure. Within a coherent modelling framework, this approach tackles the variation of neutron RBE in a wide energy range, from thermal neutrons to neutrons of hundreds of GeV, and reproduces effects related to depth in the human-sized receptor, as well as to the receptor size itself. Besides providing a better mechanistic understanding of neutron biological effectiveness, the new model can support better-informed decisions for radiation protection: indeed, current neutron weighting (ICRP)/quality (U.S. NRC) factors might be insufficient for use in some radiation protection applications, because they do not account for depth. RBE predictions obtained with the reported model were successfully compared to the currently adopted radiation protection standards when the depth information is not relevant (at the shallowest depth in the phantom or for very high energy neutrons). However, our results demonstrate that great care is needed when applying weighting factors as a function of incident neutron energy only, not explicitly considering RBE variation in the target. Finally, to facilitate the use of our results, we propose look-up RBE tables, explicitly considering the depth variable, and an analytical representation of the maximal RBE vs. neutron energy.
Quantitative genomic mapping of DNA damage may provide insights into the underlying mechanisms of damage and repair. Sequencing based approaches are bound to the limitations of PCR amplification bias and read length which hamper both the accurate quantitation of damage events and the ability to map them to structurally complex genomic regions. Optical Genome mapping in arrays of parallel nanochannels allows physical extension and genetic profiling of millions of long genomic DNA fragments, and has matured to clinical utility for characterization of complex structural aberrations in cancer genomes. Here we present a new mapping modality, Repair-Assisted Damage Detection - Optical Genome Mapping (RADD-OGM), a method for single-molecule level mapping of DNA damage on a genome-wide scale. Leveraging ultra-long reads to assemble the complex structure of a sarcoma cell-line genome, we mapped the genomic distribution of oxidative DNA damage, identifying regions more susceptible to DNA oxidation. We also investigated DNA repair by allowing cells to repair chemically induced DNA damage, pinpointing locations of concentrated repair activity, and highlighting variations in repair efficiency. Our results showcase the potential of the method for toxicogenomic studies, mapping the effect of DNA damaging agents such as drugs and radiation, as well as following specific DNA repair pathways by selective induction of DNA damage. The facile integration with optical genome mapping enables performing such analyses even in highly rearranged genomes such as those common in many cancers, a challenging task for sequencing-based approaches.
- MeSH
- Bromates toxicity MeSH
- Humans MeSH
- Chromosome Mapping * instrumentation methods MeSH
- Microfluidic Analytical Techniques * instrumentation methods MeSH
- Cell Line, Tumor MeSH
- Nanotechnology * instrumentation methods MeSH
- DNA Repair genetics MeSH
- Oxidative Stress drug effects genetics MeSH
- DNA Damage * genetics MeSH
- Gene Expression Regulation MeSH
- Gene Expression Profiling MeSH
- Toxicogenetics * instrumentation methods MeSH
- DNA Copy Number Variations MeSH
- Single Molecule Imaging * instrumentation methods MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Východiska: V tradičním pojetí vnímání univerzit byl dlouhou dobu akcentován model první a druhé role. První role univerzit byla vždy spjata se samotným vzděláváním. Druhá role pak s vědecko-výzkumnou činností. Třetí role univerzit je dnes již zakotvena jako nedílná součást dlouhodobých záměrů většiny univerzit. Hlavní myšlenkou je především skutečnost, že univerzity, fakulty a její parciální pracoviště by se neměly uzavírat do sebe samých, ale měly by zaujmout aktivní roli ve veřejném prostoru. V současnosti jedním z modelů naplňování třetí role je pojetí univerzity, která může být pojímána nebo se může profilovat jako podnikatelský subjekt – triple helix model. Tímto může docházet k větší propojenosti univerzit s aplikační a podnikatelskou praxí. Tato skutečnost následně umožňuje komercializovat výsledky vědecko-výzkumné činnosti, podílet se na pokrytí nákladových položek a případně generovat zisk konkrétních univerzitních pracovišť. Cíle: Cílem práce je představení konceptu Průmysl 4.0, jeho vliv a přesahy do oblasti práce a vzdělávání, včetně naplňování třetí role univerzit. V analytické části je cílem představit konkrétní příklady „dobré praxe“ v realizaci naplňování třetí role v rámci činnosti univerzitního vědeckotechnického parku a ve spolupráci s dalšími významnými univerzitními pracovišti v ČR a subjekty z podnikatelského prostředí. Závěry: Předložená práce naznačuje možnosti propojení univerzitního prostředí a vybraných pracovišť s podnikatelským prostředím na konkrétních příkladech dobré praxe. Jsou předloženy možnosti vzájemné spolupráce různých univerzitních pracovišť s komerční sférou, ve snaze naplňovat třetí roli univerzit v kontextu modelu triple helix.
Background: The traditional perception of universities has long emphasised the first and second role model. The first role of universities has always been linked to education itself. The second role has been linked to scientific research. The third role of universities is now embedded as an integral part of the long-term goals of most universities. Above all, the main idea is that universities, faculties and their departments should not be self-contained but should take an active role in the public sphere. Currently, one model for fulfilling the third role is the notion of a university that can be conceived or profile itself as a business entity - the triple helix model. This may lead to greater interconnection between universities and application and business practice. This in turn enables the commercialization of the results of scientific research activities, participation in cost recovery and possibly profit generation for specific university departments. Objective: The aim of the thesis is to introduce the concept of Industry 4.0, its impact and overlaps in the field of work and education, including the fulfilment of the third role of universities. In the analytical part, the aim is to present concrete examples of "good practice" in the implementation of the third role within the activities of the university science and technology park and in cooperation with other major universities in the Czech Republic and entities from the business environment. Conclusions: The presented work indicates the possibilities of linking the university environment and selected departments with the business environment on specific examples of good practice. The possibilities of mutual cooperation between different university departments and the commercial sphere are presented, in an attempt to fulfil the third role of universities in the context of the triple helix model.
SCOPE: This multi-omic study investigates the bidirectional interactions between gut microbiota and silymarin metabolism, highlighting the differential effects across various age groups. Silymarin, the extract from Silybum marianum (milk thistle), is commonly used for its hepatoprotective effects. METHODS AND RESULTS: An in vitro fermentation colon model was used with microbiota from 20 stool samples obtained from healthy donors divided into two age groups. A combination of three analytical advanced techniques, namely proton nuclear magnetic resonance (1H NMR), next-generation sequencing (NGS), and liquid chromatography-mass spectrometry (LC-MS) was used to determine silymarin microbial metabolites over 24 h, overall metabolome, and microbiota composition. Silymarin at a low diet-relevant dose of 50 μg mL-1 significantly altered gut microbiota metabolism, reducing short-chain fatty acid (acetate, butyrate, propionate) production, glucose utilization, and increasing alpha-diversity. Notably, the study reveals age-related differences in silymarin catabolism. Healthy elderly donors (70-80 years) exhibited a significant increase in a specific catabolite associated with Oscillibacter sp., whereas healthy young donors (12-45 years) showed a faster breakdown of silymarin components, particularly isosilybin B, which is associated with higher abundance of Faecalibacterium and Erysipelotrichaceae UCG-003. CONCLUSION: This study provides insights into microbiome functionality in metabolizing dietary flavonolignans, highlighting implications for age-specific nutritional strategies, and advancing our understanding of dietary (poly)phenol metabolism.
- MeSH
- Child MeSH
- Adult MeSH
- Feces microbiology MeSH
- Fermentation MeSH
- Colon * microbiology metabolism drug effects MeSH
- Fatty Acids, Volatile metabolism MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Silymarin * pharmacology MeSH
- Gastrointestinal Microbiome * drug effects physiology MeSH
- Healthy Volunteers MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Male MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Kontext: Poškození myokardu po nekardiálním chirurgickém výkonu (myocardial injury after non-cardiac surgery, MINS) představuje závažný problém přímo související s mortalitou do 30 dnů po výkonu. V diagnostice poškození myokardu se přednostně používají hodnoty vysoce senzitivního srdečního troponinu (high-sensitivity cardiac troponin, hs troponin), protože se v současnosti jedná o nejcitlivější a nejspecifičtější srdeční biomarker vhodný pro zjišťování MINS. Cílem této studie je analyzovat vztah mezi změnami v hodnotách hs troponinu a riziky spojenými s chirurgickým výkonem u pacientů podstupujících nekardiální chirurgické výkony, které by ukazovaly na poškození myokardu. Metoda: Naše studie je analytickou observační průřezovou studií, do níž byli zařazováni všichni pacienti pod-tupující nekardiální chirurgický výkon ve fakultní nemocnici Airlangga University Hospital. Krevní vzorky byly odebírány metodou náhodného vzorku (consecutive sampling) a hodnoty hs troponinu se stanovovaly před operací a 24 hodin po ní. Míra operačního rizika se hodnotila pomocí kritérií doporučených postupů ESAIC-ESC 2022. Statistická analýza se prováděla s použitím Spearmanova korelačního koeficientu. Výsledky: Do studie bylo zařazeno 75 pacientů, 39 mužů a 36 žen, ve věku s mediánem 54 let. Výsledky prokázaly statisticky významnou korelaci mezi hodnotami hs troponinu a mírou operačního rizika, a to jak předoperačními hodnotami, tak i pooperačními a jejich změnami. Medián hodnoty (mezikvartilové rozpětí) hs troponinu při nízkém operačním riziku před operací vs. po operaci činil 1,2 ng/l (0,0), resp. 1,2 ng/l (0,0), s mediánem rozdílu 0,0 ng/l (0,0); p < 0,001. Medián hodnoty hs troponinu při středně vysokém operačním riziku před operací vs. po operaci byl 1,2 ng/l (0,0) vs. 1,2 ng/l (0,0), s mediánem rozdílu 0,0 ng/l (1,6); p < 0,001. Medián hodnoty hs troponinu při vysokém operačním riziku před operací vs. po operaci byl 1,7 ng/l (18,8) vs. 11,9 ng/l (27,6); medián rozdílu činil 5,3 ng/l (11,4); p < 0,001. Závěr: Byla nalezena statisticky významná korelace mezi změnami hodnot hs troponinu a mírou operačního rizika a přítomností komplikací.
Background: Myocardial injury after non-cardiac surgery (MINS) is an important issue, directly related to the mortality rate within 30 days after surgery. High sensitivity cardiac troponin (HS troponin) is the preferred biomarker for diagnosing myocardial damage because it is the most sensitive and specific heart biomarker currently available, making it suitable for detecting MINS. The aim of this study is to analyse the relationship between changes in HS troponin levels and surgery-related risks in non-cardiac surgery patients as a marker of myocardial injury. Method: This study is an analytical observational cross sectional study that included all patients who underwent non-cardiac surgery in the Airlangga University Hospital. Samples were taken using the consecutive sampling method. HS troponin values were measured before surgery and 24 hours after surgery. The level of surgical risk was assessed based on the criteria from ESAIC-ESC 2022. Statistical analysis used Spear- man's correlation.
- MeSH
- Anesthesia adverse effects MeSH
- Biomarkers blood MeSH
- Surgical Procedures, Operative * adverse effects MeSH
- Adult MeSH
- Cardiomyopathies diagnosis etiology blood MeSH
- Middle Aged MeSH
- Humans MeSH
- Cross-Sectional Studies MeSH
- Risk MeSH
- Aged MeSH
- Statistics as Topic MeSH
- Troponin * blood MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Observational Study MeSH
