Lens epithelium-derived growth factor p75 (LEDGF/p75), member of the hepatoma-derived growth-factor-related protein (HRP) family, is a transcriptional co-activator and involved in several pathologies including HIV infection and malignancies such as MLL-rearranged leukemia. LEDGF/p75 acts by tethering proteins to the chromatin through its integrase binding domain. This chromatin interaction occurs between the PWWP domain of LEDGF/p75 and nucleosomes carrying a di- or trimethylation mark on histone H3 Lys36 (H3K36me2/3). Our aim is to rationally devise small molecule drugs capable of inhibiting such interaction. To bootstrap this development, we resorted to X-ray crystallography-based fragment screening (FBS-X). Given that the LEDGF PWWP domain crystals were not suitable for FBS-X, we employed crystals of the closely related PWWP domain of paralog HRP-2. As a result, as many as 68 diverse fragment hits were identified, providing a detailed sampling of the H3K36me2/3 pocket pharmacophore. Subsequent structure-guided fragment expansion in three directions yielded multiple compound series binding to the pocket, as verified through X-ray crystallography, nuclear magnetic resonance and differential scanning fluorimetry. Our best compounds have double-digit micromolar affinity and optimally sample the interactions available in the pocket, judging by the Kd-based ligand efficiency exceeding 0.5 kcal/mol per non-hydrogen atom. Beyond π-stacking within the aromatic cage of the pocket and hydrogen bonding, the best compounds engage in a σ-hole interaction between a halogen atom and a conserved water buried deep in the pocket. Notably, the binding pocket in LEDGF PWWP is considerably smaller compared to the related PWWP1 domains of NSD2 and NSD3 which feature an additional subpocket and for which nanomolar affinity compounds have been developed recently. The absence of this subpocket in LEDGF PWWP limits the attainable affinity. Additionally, these structural differences in the H3K36me2/3 pocket across the PWWP domain family translate into a distinct selectivity of the compounds we developed. Our top-ranked compounds are interacting with both homologous LEDGF and HRP-2 PWWP domains, yet they showed no affinity for the NSD2 PWWP1 and BRPF2 PWWP domains which belong to other PWWP domain subfamilies. Nevertheless, our developed compound series provide a strong foundation for future drug discovery targeting the LEDGF PWWP domain as they can further be explored through combinatorial chemistry. Given that the affinity of H3K36me2/3 nucleosomes to LEDGF/p75 is driven by interactions within the pocket as well as with the DNA-binding residues, we suggest that future compound development should target the latter region as well. Beyond drug discovery, our compounds can be employed to devise tool compounds to investigate the mechanism of LEDGF/p75 in epigenetic regulation.

• MeSH
• knihovny malých molekul chemie   farmakologie   chemická syntéza   MeSH
• krystalografie rentgenová MeSH
• lidé MeSH
• mezibuněčné signální peptidy a proteiny metabolismus   chemie   MeSH
• molekulární modely MeSH
• molekulární struktura MeSH
• proteinové domény MeSH
• racionální návrh léčiv * MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Early identification of those with NAFLD activity score ≥ 4 and significant fibrosis (≥F2) or at-risk metabolic dysfunction-associated steatohepatitis (MASH) is a priority as these patients are at increased risk for disease progression and may benefit from therapies. We developed and validated a highly specific metabolomics-driven score to identify at-risk MASH. METHODS: We included derivation (n = 790) and validation (n = 565) cohorts from international tertiary centers. Patients underwent laboratory assessment and liver biopsy for metabolic dysfunction-associated steatotic liver disease. Based on 12 lipids, body mass index, aspartate aminotransferase, and alanine aminotransferase, the MASEF score was developed to identify at-risk MASH and compared to the FibroScan-AST (FAST) score. We further compared the performance of a FIB-4 + MASEF algorithm to that of FIB-4 + liver stiffness measurements (LSM) by vibration-controlled transient elastography (VCTE). RESULTS: The diagnostic performance of the MASEF score showed an area under the receiver-operating characteristic curve, sensitivity, specificity, and positive and negative predictive values of 0.76 (95% CI 0.72-0.79), 0.69, 0.74, 0.53, and 0.85 in the derivation cohort, and 0.79 (95% CI 0.75-0.83), 0.78, 0.65, 0.48, and 0.88 in the validation cohort, while FibroScan-AST performance in the validation cohort was 0.74 (95% CI 0.68-0.79; p = 0.064), 0.58, 0.79, 0.67, and 0.73, respectively. FIB-4+MASEF showed similar overall performance compared with FIB-4 + LSM by VCTE ( p = 0.69) to identify at-risk MASH. CONCLUSION: MASEF is a promising diagnostic tool for the assessment of at-risk MASH. It could be used alternatively to LSM by VCTE in the algorithm that is currently recommended by several guidance publications.

• MeSH
• biopsie škodlivé účinky   MeSH
• elastografie * MeSH
• fibróza MeSH
• jaterní cirhóza patologie   MeSH
• játra diagnostické zobrazování   patologie   MeSH
• lidé MeSH
• nealkoholová steatóza jater * patologie   MeSH
• prediktivní hodnota testů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Monogenic diabetes is a gateway to precision medicine through molecular mechanistic insight. Hepatocyte nuclear factor 1A (HNF-1A) and HNF-4A are transcription factors that engage in crossregulatory gene transcription networks to maintain glucose-stimulated insulin secretion in pancreatic β cells. Variants in the HNF1A and HNF4A genes are associated with maturity-onset diabetes of the young (MODY). Here, we explored 4 variants in the P2-HNF4A promoter region: 3 in the HNF-1A binding site and 1 close to the site, which were identified in 63 individuals from 21 families of different MODY disease registries across Europe. Our goal was to study the disease causality for these variants and to investigate diabetes mechanisms on the molecular level. We solved a crystal structure of HNF-1A bound to the P2-HNF4A promoter and established a set of techniques to probe HNF-1A binding and transcriptional activity toward different promoter variants. We used isothermal titration calorimetry, biolayer interferometry, x-ray crystallography, and transactivation assays, which revealed changes in HNF-1A binding or transcriptional activities for all 4 P2-HNF4A variants. Our results suggest distinct disease mechanisms of the promoter variants, which can be correlated with clinical phenotype, such as age of diagnosis of diabetes, and be important tools for clinical utility in precision medicine.

• MeSH
• diabetes mellitus 2. typu * genetika   metabolismus   MeSH
• hepatocytární jaderný faktor 1-alfa * genetika   metabolismus   MeSH
• hepatocytární jaderný faktor 4 * genetika   metabolismus   MeSH
• krystalografie rentgenová MeSH
• lidé MeSH
• promotorové oblasti (genetika) * genetika   MeSH
• regulace genové exprese MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Advances in single-cell level analytical techniques, especially cytometric approaches, have led to profound innovation in biomedical research, particularly in the field of clinical immunology. This has resulted in an expansion of high-dimensional data, posing great challenges for comprehensive and unbiased analysis. Conventional manual analysis is thus becoming untenable to handle these challenges. Furthermore, most newly developed computational methods lack flexibility and interoperability, hampering their accessibility and usability. Here, we adapted Seurat, an R package originally developed for single-cell RNA sequencing (scRNA-seq) analysis, for high-dimensional flow cytometric data analysis. Based on a 20-marker antibody panel and analyses of T-cell profiles in both adult blood and cord blood (CB), we showcased the robust capacity of Seurat in flow cytometric data analysis, which was further validated by Spectre, another high-dimensional cytometric data analysis package, and conventional manual analysis. Importantly, we identified a unique CD8+ T-cell population defined as CD8+CD45RA+CD27+CD161+ T cell that was predominantly present in CB. We characterised its IFN-γ-producing and potential cytotoxic properties using flow cytometry experiments and scRNA-seq analysis from a published dataset. Collectively, we identified a unique human CB CD8+CD45RA+CD27+CD161+ T-cell subset and demonstrated that Seurat, a widely used package for scRNA-seq analysis, possesses great potential to be repurposed for cytometric data analysis. This facilitates an unbiased and thorough interpretation of complicated high-dimensional data using a single analytical pipeline and opens a novel avenue for data-driven investigation in clinical immunology.

• MeSH
• analýza jednotlivých buněk metody   MeSH
• antigeny CD27 metabolismus   imunologie   MeSH
• antigeny CD45 * metabolismus   imunologie   MeSH
• CD8-pozitivní T-lymfocyty * imunologie   MeSH
• dospělí MeSH
• fetální krev * imunologie   cytologie   MeSH
• imunofenotypizace metody   MeSH
• lektinové receptory NK-buněk - podrodina B imunologie   metabolismus   MeSH
• lidé MeSH
• průtoková cytometrie * metody   MeSH
• software MeSH
• T-lymfocyty - podskupiny imunologie   metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Worldwide stroke is the second leading cause of death and the third leading cause of death and disability combined. The estimated global economic burden by stroke is over US$891 billion per year. Within three decades (1990-2019), the incidence increased by 70%, deaths by 43%, prevalence by 102%, and DALYs by 143%. Of over 100 million people affected by stroke, about 76% are ischemic stroke (IS) patients recorded worldwide. Contextually, ischemic stroke moves into particular focus of multi-professional groups including researchers, healthcare industry, economists, and policy-makers. Risk factors of ischemic stroke demonstrate sufficient space for cost-effective prevention interventions in primary (suboptimal health) and secondary (clinically manifested collateral disorders contributing to stroke risks) care. These risks are interrelated. For example, sedentary lifestyle and toxic environment both cause mitochondrial stress, systemic low-grade inflammation and accelerated ageing; inflammageing is a low-grade inflammation associated with accelerated ageing and poor stroke outcomes. Stress overload, decreased mitochondrial bioenergetics and hypomagnesaemia are associated with systemic vasospasm and ischemic lesions in heart and brain of all age groups including teenagers. Imbalanced dietary patterns poor in folate but rich in red and processed meat, refined grains, and sugary beverages are associated with hyperhomocysteinaemia, systemic inflammation, small vessel disease, and increased IS risks. Ongoing 3PM research towards vulnerable groups in the population promoted by the European Association for Predictive, Preventive and Personalised Medicine (EPMA) demonstrates promising results for the holistic patient-friendly non-invasive approach utilising tear fluid-based health risk assessment, mitochondria as a vital biosensor and AI-based multi-professional data interpretation as reported here by the EPMA expert group. Collected data demonstrate that IS-relevant risks and corresponding molecular pathways are interrelated. For examples, there is an evident overlap between molecular patterns involved in IS and diabetic retinopathy as an early indicator of IS risk in diabetic patients. Just to exemplify some of them such as the 5-aminolevulinic acid/pathway, which are also characteristic for an altered mitophagy patterns, insomnia, stress regulation and modulation of microbiota-gut-brain crosstalk. Further, ceramides are considered mediators of oxidative stress and inflammation in cardiometabolic disease, negatively affecting mitochondrial respiratory chain function and fission/fusion activity, altered sleep-wake behaviour, vascular stiffness and remodelling. Xanthine/pathway regulation is involved in mitochondrial homeostasis and stress-driven anxiety-like behaviour as well as molecular mechanisms of arterial stiffness. In order to assess individual health risks, an application of machine learning (AI tool) is essential for an accurate data interpretation performed by the multiparametric analysis. Aspects presented in the paper include the needs of young populations and elderly, personalised risk assessment in primary and secondary care, cost-efficacy, application of innovative technologies and screening programmes, advanced education measures for professionals and general population-all are essential pillars for the paradigm change from reactive medical services to 3PM in the overall IS management promoted by the EPMA.

• Publikační typ
• časopisecké články MeSH

Among medical specialties, laboratory medicine is the largest producer of structured data and must play a crucial role for the efficient and safe implementation of big data and artificial intelligence in healthcare. The area of personalized therapies and precision medicine has now arrived, with huge data sets not only used for experimental and research approaches, but also in the "real world". Analysis of real world data requires development of legal, procedural and technical infrastructure. The integration of all clinical data sets for any given patient is important and necessary in order to develop a patient-centered treatment approach. Data-driven research comes with its own challenges and solutions. The Findability, Accessibility, Interoperability, and Reusability (FAIR) Guiding Principles provide guidelines to make data findable, accessible, interoperable and reusable to the research community. Federated learning, standards and ontologies are useful to improve robustness of artificial intelligence algorithms working on big data and to increase trust in these algorithms. When dealing with big data, the univariate statistical approach changes to multivariate statistical methods significantly shifting the potential of big data. Combining multiple omics gives previously unsuspected information and provides understanding of scientific questions, an approach which is also called the systems biology approach. Big data and artificial intelligence also offer opportunities for laboratories and the In Vitro Diagnostic industry to optimize the productivity of the laboratory, the quality of laboratory results and ultimately patient outcomes, through tools such as predictive maintenance and "moving average" based on the aggregate of patient results.

• MeSH
• algoritmy MeSH
• big data * MeSH
• individualizovaná medicína metody   MeSH
• lidé MeSH
• poskytování zdravotní péče MeSH
• umělá inteligence * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Úvod: Plicní resekce může zapříčinit změny ve funkčním stavu a z toho důvodu také v kvalitě života pacientů, kteří ji podstupují. Aby bylo plánování péče o tyto pacienty efektivní, je potřeba identifikovat oblasti, které si vyžadují zvláštní pozornost. Cíl: Cílem této prospektivní studie bylo porovnat kvalitu života a disabilitu před plicní resekcí a tři měsíce po ní. Metodika: Kvalita života a disabilita byly hodnoceny pomocí standardizovaných nástrojů: Short Form-12 verze 2 a WHO Disability Assessment Schedule 2. Výsledky: Výzkumný vzorek zahrnoval 44 pacientů indikovaných k plicní resekci. Průměrný věk v souboru byl 65,7 let (min. 37, max. 79), 56,8 % představovali muži. Tři měsíce po resekci bylo zjištěno zhoršení kvality života v doménách Fyzické zdraví (p = 0,016) a Bolest (p = 0.044). Došlo také k signifikantní změně celkové úrovně disability (p < 0.001). Signifikantní změny disability byly zjištěny také v doménách Mobilita (p < 0.001), Životní aktivity (p < 0.001) a Účast ve společnosti (p = 0.037). Závěr: Tři měsíce po resekci plic bylo zjištěno statisticky významné zhoršení kvality života a disability ve více doménách. Fyzické zdraví, Bolest, Mobilita, Životní aktivity a Účast ve společnosti byly identifikovány jako oblasti, které při plánování pooperační péče o tyto pacienty vyžadují zvláštní pozornost.

Background: Lung resection can cause changes in patients' functional status, potentially affecting their quality of life. For effective care planning after resection, it is necessary to identify areas of quality of life that require special attention. Objectives: This prospective study aimed to compare the quality of life, health, and disability before lung resection and three months after it. Methods: Quality of life, health, and disability were assessed using standardized tools: Short Form-12 version 2, and WHO Disability Assessment Schedule 2. Results: The research group included 44 patients with a mean age of 65.7 years (min. 37, max. 79) referred for lung resection. Men were more represented in the study group (56.8%) than women. Three months after the surgery, statistically significant deterioration in the quality of life was observed in the domains of Physical functioning (p = 0.016) and Bodily pain (p = 0.044). Where disability was concerned, the overall score also deteriorated (p < 0.001). This was driven predominantly by the domains of Mobility (p < 0.001), Life activities (p < 0.001), and Participation (p = 0.037). Conclusion: The deterioration in the quality of life in multiple domains three months after lung resection is highly significant. The deterioration in the domains Physical functioning and Bodily pain in the quality of life field, and of Mobility, Life activities, and Participation in the health and disability score identifies these areas as those that need special attention when planning post-surgery care for these patients.

• MeSH
• kvalita života * MeSH
• lidé MeSH
• pneumektomie * MeSH
• prospektivní studie MeSH
• Check Tag
• lidé MeSH

OBJECTIVES: Multidisciplinary teams in cancer care are increasingly using information and communication technology (ICT), hospital health information system (HIS) functionalities and ICT-driven care components. We aimed to explore the use of these tools in multidisciplinary team meetings (MTMs) and to identify the critical challenges posed by their adoption based on the perspective of professionals representatives from European scientific societies. DESIGN: This qualitative study used discussion of cases and focus group technique to generate data. Thematic analysis was applied. SETTING: Healthcare professionals working in a multidisciplinary cancer care environment. PARTICIPANTS: Selection of informants was carried out by European scientific societies in accordance with professionals' degree of experience in adopting the implementation of ICT and from different health systems. RESULTS: Professionals representatives of 9 European scientific societies were involved. Up to 10 ICTs, HIS functionalities and care components are embedded in the informational and decision-making processes along three stages of MTMs. ICTs play a key role in opening MTMs to other institutions (eg, by means of molecular tumour boards) and information types (eg, patient-reported outcome measures), and in contributing to the internal efficiency of teams. While ICTs and care components have their own challenges, the information technology context is characterised by the massive generation of unstructured data, the lack of interoperability between systems from different hospitals and HIS that are conceived to store and classify information rather than to work with it. CONCLUSIONS: The emergence of an MTM model that is better integrated in the wider health system context and incorporates inputs from patients and support systems make traditional meetings more dynamic and interconnected. Although these changes signal a second transition in the development process of multidisciplinary teams, they occur in a context marked by clear gaps between the information and management needs of MTMs and the adequacy of current HIS.

• MeSH
• informační technologie * MeSH
• komunikace MeSH
• lidé MeSH
• nádory * terapie   MeSH
• poskytování zdravotní péče MeSH
• týmová péče o pacienty MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Introduction: Cardiorespiratory fitness (CRF) testing is routinely performed by most armed and tactical forces around the world as part of their recruitment process for new members or simply as an annual examination of their personnel. A 2-mile run (2MR) test is among the most commonly used. However, as it is performed outdoors, weather, climate, and terrain can influence the results and often limit the maximum performance. Also, this test has been shown to be difficult for individuals because the pacing strategy is self-paced. As such, it does not reflect the real situation on the battlefield, where most activities are externally driven by the environment and the enemy. Therefore, we believe that the 30-15 Intermittent Fitness Test (30-15IFT) may be a suitable tool for measuring CRF and battle readiness of military personnel. Moreover, given the importance of visual attention to military personnel we aim to investigate the differences in visual attention between better and less physically prepared infantry members and its corresponding response to maximal endurance running test. Methods and analysis: This randomized cross-over study using a within-subjects test-retest design will enroll 32 infantry members of the Slovenian Armed Forces. To investigate the reliability and validity of the 30-15IFT test, an incremental treadmill test (TR), a 2MR test, and two identical 30-15IFT will be performed in randomized order. Additionally, participants will be subsequently divided into two groups based on their score on the Army Physical Fitness Test (APFT), whereas differences in visual attention will be assessed by using the d2 test. The primary analysis will assess differences in key physiological outcomes between the different CRF tests (TR vs. 2MR vs. 30-15IFT). In addition, the relative reliability of all dependent variables between two 30-15IFT trials will be estimated by interclass correlation coefficient (ICC), while relationship between maximal oxygen uptake, heart rate and maximal running speed of 30-15IFT, TR and 2MR will be assessed using Pearson's correlation. Ethics and dissemination: Ethical approval was obtained from the National Medical Ethics Committee (reference number: 0120-495/2021/6). The results of the proposed study will be disseminated through publication in peer-reviewed journals. Clinical Trial Registration: [www.ClinicalTrials.gov], identifier [NCT05218798].

• Publikační typ
• časopisecké články MeSH

Epidural spinal cord stimulation (ESCS) is widely used for chronic pain treatment, and is also a promising tool for restoring motor function after spinal cord injury. Despite significant positive impact of ESCS, currently available protocols provide limited specificity and efficiency partially due to the limited number of contacts of the leads and to the limited flexibility to vary the spatial distribution of the stimulation field in respect to the spinal cord. Recently, we introduced Orientation Selective (OS) stimulation strategies for deep brain stimulation, and demonstrated their selectivity in rats using functional MRI (fMRI). The method achieves orientation selectivity by controlling the main direction of the electric field gradients using individually driven channels. Here, we introduced a similar OS approach for ESCS, and demonstrated orientation dependent brain activations as detected by brain fMRI. The fMRI activation patterns during spinal cord stimulation demonstrated the complexity of brain networks stimulated by OS-ESCS paradigms, involving brain areas responsible for the transmission of the motor and sensory information. The OS approach may allow targeting ESCS to spinal fibers of different orientations, ultimately making stimulation less dependent on the precision of the electrode implantation.

• MeSH
• epidurální prostor diagnostické zobrazování   patofyziologie   MeSH
• implantované elektrody MeSH
• krysa rodu rattus MeSH
• magnetická rezonanční tomografie * MeSH
• míšní stimulace * MeSH
• poranění míchy * diagnostické zobrazování   patofyziologie   terapie   MeSH
• potkani Sprague-Dawley MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH