Search Results (279)

T cell receptor (TCR) profiling using next-generation sequencing (NGS) enables high-throughput, in-depth analysis of repertoire diversity, offering numerous clinical applications. We developed a DNA-based strategy to analyse the TCRβ-chain using NGS and established reference values for T cell repertoire characteristics in 74 healthy donors, including 44 adults, 20 paediatrics, and 10 cord blood units (CBUs). Additionally, four paediatric patients with combined immunodeficiency (CID) or severe CID (SCID) due to deleterious mutations in recombination activating genes (RAG) were analysed. The developed strategy demonstrated high specificity, reproducibility, and sensitivity, and all functional variable and joining genes were detected with minimal PCR bias. All donors had a Gaussian-like distribution of complementary-determining region 3 length, with lower presence of non-templated nucleotides and higher proportion of non-functional clonotypes in CBUs. Both CBUs and paediatrics showed greater convergence and TCRβ diversity was significantly lower in adults and donors with cytomegalovirus-positive serostatus. Finally, an analysis of paediatric patients with RAG-SCID/CID showed significantly shorter CDR3 region length and lower repertoire diversity compared to healthy paediatrics. In summary, we developed a reliable and feasible TCRβ sequencing strategy for application in the clinical setting, and established reference values that could assist in the diagnosis and monitoring of pathological conditions affecting the T cell repertoire. Full article

This study aimed to evaluate the impact of an 8-week reverse Nordic exercise training (RNET) program on physical fitness in male youth soccer players. A total of 35 players participated in the study and were divided into two groups: the RNET group (n = 19, age 16.39 ± 0.46 years) and the active control group (CG: n = 16, age 16.53 ± 0.48 years). To assess fitness changes, participants were tested on linear sprint speed (5, 10, and 20 m sprints), change-of-direction (CiD) speed (505-CiD), vertical jump (countermovement jump [CMJ]), horizontal jump (standing long jump [SLJ]), drop jump (20 cm drop jump [DJ-20]), and repeated sprint ability (RSA). Significant group-by-time interactions were observed (effect size, [ES] = 0.70 to 1.37), with substantial improvements in the RNET group across linear sprint, CiD, and jumping performances (ES = 0.61 to 1.47), while no significant changes were noted in the CG. However, no significant group-by-time interactions were observed for RSA parameters. Individual response analysis revealed that 63–89% of RNET group exhibited improvements exceeding the smallest worthwhile change (SWC_0.2) threshold. These results suggest that the RNET program is both effective and safe for enhancing physical fitness in male youth soccer players. Full article

Cid1 protein is a crucial component in the RNA interference pathway and abnormal nuclear RNA turnover processes, primarily responsible for adding uridine to the 3′ end of RNA. Cid1 exhibits selective polymerization of UTP over other nucleoside triphosphates. To explore the mechanism of this selectivity, five systems: free-Cid1, Cid1-ATP, Cid1-UTP, Cid1-CTP, and Cid1-GTP with 500 ns Gaussian accelerated molecular dynamics (GaMD) simulations were performed to investigate conformational changes and binding affinities between substrates and Cid1. The results showed that UTP formed stronger and more numerous non-covalent interactions with Cid1 compared to the other three substrates. The Molecular Mechanics Poisson-Boltzmann Surface Area (MM-PBSA) binding energy analysis revealed a substrate preference for Cid1 polymerase in the order of UTP, followed by ATP, CTP, and GTP. These findings provide theoretical insights into the substrate selectivity mechanism of Cid1 and provide theoretical clues for the design and modification of Cid1 polymerase. Full article

Background: Some patients with juvenile idiopathic arthritis (JIA) can successfully undergo withdrawal of treatment with anti-tumor necrosis factor alpha (anti-TNF-α) therapy, which may reduce economic and treatment-related burdens and the potential morbidity of treatment for at least 6 months. Currently, no guidelines exist on the appropriate withdrawal of anti-TNF-α therapy once clinically inactive disease (CID) has been achieved. This study aimed to assess the possibility of withdrawing anti-TNF-α therapy in children with non-systemic JIA after achieving long-term clinical remission. Methods: This single-center retrospective cohort study included data from 137 non-systemic JIA patients treated with anti-TNF-α therapy (etanercept or adalimumab) and having maintained CID for at least 24 months during treatment. Demographic, laboratory, and treatment data were collected at JIA onset, at the initiation of anti-TNF-α therapy, every 6 months during therapy, and at the time of disease flare. Anti-TNF-α therapy was discontinued abruptly after discussing it with the patients and their families in each case. Outcomes were assessed using standard criteria for remission in JIA (ACRpedi and Wallace criteria). Results: Following withdrawal of TNF-α inhibitors, 93/137 patients (67.9%) experienced a disease flare, with a median time to flare of 7 months (3; 14). Thirty-two percent of patients remained in remission for a median of 63 months. Absence of flare during the first 22 months after discontinuation was associated with prolonged biologic-free remission (odds ratio 682; 95% CI 38.6–12,062; p < 0.0001), with 83% sensitivity and 100% specificity (area under the ROC curve 0.967). Most flares involved arthritis (76%) and/or uveitis (24%), primarily affecting knees, ankles, and wrists. Inflammatory markers were generally lower at flare compared to baseline. Biological therapy was resumed in 84/93 patients (90.3%), achieving at least a 50% improvement according to ACRpedi criteria within 3 months and remission according to Wallace criteria within 6 months. Conclusion: Over two-thirds of patients with non-systemic JIA who achieve CID experience a flare within seven months of anti-TNF-α discontinuation. Re-initiation of biologic therapy is effective in restoring remission. These results indicate that prolonged biologic-free remission is possible in a subset of patients, highlighting the need for individualized withdrawal strategies and careful post-discontinuation monitoring. Full article

Arachidonic acid (eicosatetraenoic acid) is the precursor of the eicosanoids, which include prostaglandins (PG). Methods based on GC-MS/MS are the Gold Standard for the quantitative analysis of eicosanoids in biological samples. After extraction and derivatization, biological F₂-prostaglandins are analyzed on quadrupole GC-MS/MS apparatus as pentafluorobenzyl (PFB) ester trimethylsilyl (TMS) ether derivatives, i.e., PFB-TMS. Negative-ion chemical ionization (NICI) in the ion source generates abundant anions due to [M-PFB]⁻, which are detected in the selected ion monitoring (SIM) mode. Collision-induced dissociation (CID) of [M-PFB]⁻ in the collision cell generates numerous product ions, which are suitable candidates for quantitative analyses in the selected reaction monitoring (SRM) mode. In this article, we report on investigations of gas-phase reactions of PFB-TMS derivatives of F₂-prostaglandins, which consist of PGF_2α, 8-iso-PGF_2α, and up to 62 further isomers, known as the F₂-isoprostanes. We performed a meta-analysis of previously reported CID mass spectra (32 eV) of PFB-(TMS)₃ of seven chemically closely related isomeric F₂-prostaglandins of the 15-F_2t-IsoP type. This unique dataset contains 19 product ions generated by CID of the common precursor at m/z 569 [M-PFB]⁻ in the m/z range of 150–600. All isomers produced the same product ions, which, however, greatly differed in their intensity. Principal Component Analysis (PCA) and Receiver Operating Characteristic (ROC) Analysis (ROCA) were performed. Two compounds, i.e., 8-iso-9β,11α-PGF_2α and 9α,11β-PGF_2α, and two product ions, i.e., m/z 299 [M-PFB-3×TMSOH]⁻ and m/z 215 [M-PFB-3×TMSOH-C₄H₈-C₂H₄]⁻, were noticeable. ROCA revealed the highest disagreement between PGF_2α and 8-iso-9β,11α-PGF_2α (AUC = 0.7075 ± 0.0834, p = 0.0248). PCA and ROCA are of limited value in the GC-MS/MS of closely chemically related F₂-prostaglandins. Fragmentation mechanisms were proposed for the formation of all 19 product ions generated by CID of common precursor anions due to [M-PFB]⁻. Full article

Diffuse astrocytoma is a primary brain tumor known for its gradual and diffuse infiltration into the surrounding brain tissue. Given this characteristic, the investigation of the peritumoral region holds potential biological and clinical relevance. In this study, ion mobility spectrometry mass spectrometry (IMS MS) was optimized and applied for the first time for the analysis of gangliosides present in the peritumoral tissue of diffuse astrocytoma. Ganglioside profiling and structural characterization were conducted using high-resolution nanoelectrospray ionization (nanoESI) IMS MS, along with tandem mass spectrometry (MS/MS) via low-energy collision-induced dissociation (CID) in the negative ion mode. Using IMS MS-based separation and screening, we observed a greater diversity of ganglioside species in the peritumoral tissue than previously reported. Notably, an elevated expression was detected for several species, including GT1(d18:1/18:0), GT1(d18:1/20:0), GM2(d18:1/16:2), GD1(d18:1/16:0), GD2(d18:1/20:0), Fuc-GT3(d18:1/24:4), and Fuc-GD1(d18:1/18:2). Although preliminary, these observations prompt consideration of whether these species could be implicated in processes such as microenvironmental modulation, tumor cell infiltration and invasion, maintenance of cellular interactions, or regulation of immune responses. Additionally, their potential utility as biomarkers may merit further exploration. In the subsequent phase of the study, structural analysis using IMS MS, CID tandem MS, and fragmentation data supported the identification of GT1b(d18:1/20:0) isomer in the peritumoral tissue. However, given the exploratory nature of the study and reliance on limited sampling, further investigation across broader sample sets is necessary to extend these findings. Full article

Cyclophilins (Cyps), a family of peptidyl-prolyl isomerases, play essential roles in the life cycle of coronaviruses by interacting with viral proteins and modulating host immune responses. In this systematic review, we examined cell culture, animal model, and clinical studies assessing the anti-viral efficacy of cyclosporine A (CsA, PubChem CID: 5284373) and its non-immunosuppressive derivatives against coronaviruses. CsA demonstrated robust anti-viral activity in vitro across a broad range of coronaviruses, including but not limited to HCoV-229E, SARS-CoV, MERS-CoV, and SARS-CoV-2, with potent EC₅₀ values in the low micromolar range. Non-immunosuppressive analogs such as Alisporivir and NIM811 exhibited similar inhibitory effects. In vivo, CsA treatment significantly reduced viral load, ameliorated lung pathology, and improved survival in coronavirus-infected animals. Clinical studies further indicated that CsA administration was associated with improved outcomes in COVID-19 patients, including reduced mortality and shorter hospital stays. Mechanistic studies revealed that CsA disrupts the formation of viral replication complexes, interferes with critical Cyp–viral protein interactions, and modulates innate immune signaling. These findings collectively demonstrate the therapeutic potential of cyclophilin inhibitors as broad-spectrum anti-virals against current and emerging coronaviruses. Full article

The global climate crisis has driven unprecedented agreements among nations on carbon mitigation. With China’s commitment to carbon peaking and carbon neutrality targets, the building sector has emerged as a critical focus for emission reduction, particularly because office buildings account for over 30% of building energy consumption. However, a systematic and regionally adaptive low-carbon technology evaluation framework is lacking. To address this gap, this study develops a multidimensional decision-making system to quantify and rank low-carbon technologies for office buildings in Beijing. The method includes four core components: (1) establishing three archetypal models—low-rise (H ≤ 24 m), mid-rise (24 m < H ≤ 50 m), and high-rise (50 m < H ≤ 100 m) office buildings—based on 99 office buildings in Beijing; (2) classifying 19 key technologies into three clusters—Envelope Structure Optimization, Equipment Efficiency Enhancement, and Renewable Energy Utilization—using bibliometric analysis and policy norm screening; (3) developing a four-dimensional evaluation framework encompassing Carbon Reduction Degree (CRD), Economic Viability Degree (EVD), Technical Applicability Degree (TAD), and Carbon Intensity Degree (CID); and (4) conducting a comprehensive quantitative evaluation using the AHP-entropy-TOPSIS algorithm. The results indicate distinct priority patterns across the building types: low-rise buildings prioritize roof-mounted photovoltaic (PV) systems, LED lighting, and thermal-break aluminum frames with low-E double-glazed laminated glass. Mid- and high-rise buildings emphasize integrated PV-LED-T8 lighting solutions and optimized building envelope structures. Ranking analysis further highlights LED lighting, T8 high-efficiency fluorescent lamps, and rooftop PV systems as the top-recommended technologies for Beijing. Additionally, four policy recommendations are proposed to facilitate the large-scale implementation of the program. This study presents a holistic technical integration strategy that simultaneously enhances the technological performance, economic viability, and carbon reduction outcomes of architectural design and renovation. It also establishes a replicable decision-support framework for decarbonizing office and public buildings in cities, thereby supporting China’s “dual carbon” goals and contributing to global carbon mitigation efforts in the building sector. Full article

Objectives: This randomized controlled trial investigated the effects of an 8-week, volume-equated eccentric training program comprising Nordic hamstring and reverse Nordic exercises performed either once or twice per week on measures of physical fitness in pubertal male soccer players. Methods: A total of 34 participants were randomly assigned into a 1-day (n = 16; age = 14.58 ± 0.28 years) or 2-day (n = 18; age = 14.84 ± 0.22 years) per week training group. Physical fitness was assessed using 5 m and 10 m sprints, the 505 change in direction (CiD) speed test, Y-shaped agility test, countermovement jump (CMJ), and standing long jump (SLJ). Results: Significant group-by-time interactions were observed for the 505 CiD test, agility, and CMJ performance (effect sizes [ES] = 0.80 to 1.13). However, no significant interactions were found for the 5 and 10 m sprints or for SLJ (p > 0.05). Compared to the 1-day group, the 2-day training group showed greater improvements in CiD speed (∆7.36%; p < 0.001; ES = 0.92), agility (∆7.91%; p < 0.001; ES = 1.68), and CMJ (∆7.44%; p < 0.01; ES = 0.35), while no differences were observed in 5 and 10 m linear sprints or SLJ performance. According to individual response analysis, improvements across the physical fitness parameters beyond the smallest worthwhile change (SWC_0.2) were observed in 22–83% of players in the 1-day group and 77–100% in the 2-day group. Conclusions: In summary, the findings suggest that when training volume is matched, distributing the eccentric training regimen over two days per week may lead to greater improvements in CiD speed, agility, and CMJ performance compared to a single-day approach. Full article

The Chikungunya virus (CHIKV) continues to pose a significant global health challenge due to the absence of effective antiviral treatments and limited vaccine availability. This study employed a comprehensive in silico workflow, incorporating high-throughput virtual screening, binding free-energy calculations, ADMET (absorption, distribution, metabolism, excretion, and toxicity) analysis, and 200 ns molecular dynamics (MD) simulations, to identify new inhibitors targeting the E1–E2 glycoprotein complex, crucial for CHIKV entry and membrane fusion. Four promising candidates were identified from a library of 20,000 compounds, with CID 136801451 showing the most potent binding (docking score: −10.227; ΔG_bind: −51.53 kcal/mol). The top four compounds exhibited favorable ADMET profiles, meeting nearly all criteria. MD simulations confirmed stable binding and strong interactions between CID 136801451 and the E1–E2 complex, evidenced by consistently low RMSD values. These findings highlight CID 136801451 as a promising CHIKV entry inhibitor, warranting further in vitro and in vivo evaluation to advance the development of effective anti-CHIKV therapeutics. Full article

The avian influenza virus, particularly the highly pathogenic H5N1 subtype, represents a significant public health threat due to its interspecies transmission potential and growing resistance to current antiviral therapies. To address this, the identification of novel and effective neuraminidase (NA) inhibitors is critical. In this study, an integrated in silico strategy was employed, beginning with the generation of an energy-optimized pharmacophore model (e-pharmacophore, ADDN) based on the reference inhibitor Zanamivir. A virtual screening of 47,781 natural compounds from the PubChem database was performed, followed by molecular docking validated through an enrichment assay. Promising hits were further evaluated via ADMET predictions, density functional theory (DFT) calculations to assess chemical reactivity, and molecular dynamics (MD) simulations to examine the stability of the ligand–protein complexes. Three lead compounds (C1: CID 102209473, C2: CID 85692821, and C3: CID 45379525) demonstrated strong binding affinity toward NA. Their ADMET profiles predicted favorable bioavailability and low toxicity. The DFT analyses indicated suitable chemical reactivity, particularly for C2 and C3. The MD simulations confirmed the structural stability of all three ligand–NA complexes, supported by robust and complementary intermolecular interactions. In contrast, Zanamivir exhibited limited hydrophobic interactions, compromising its binding stability within the active site. These findings offer a rational foundation for further experimental validation and the development of next-generation NA inhibitors derived from natural sources. Full article

Acute Myeloid Leukemia (LAML) is a life-threatening hematological malignancy, and the DEAD-box helicase 3 X-linked (DDX3X) gene is a potential yet underexplored target gene for LAML. Biomolecules derived from medicinal plants like Aerva javanica offer a great source of therapeutic candidates. This study aimed to investigate the role of DDX3X in LAML and identify plant-derived biomolecules that could inhibit DDX3X using computational approaches. Pan-cancer mutational profiling, a transcriptomic analysis, survival, protein–protein interaction networks, and a principal component analysis (PCA) were employed to elucidate functional associations and transcriptomic divergence. Subsequently, biomolecules from A. javanica were subjected to in silico screening using molecular docking and ADMET profiling. The docking protocol was validated using RK-33, a known DDX3X inhibitor. DDX3X was found to be linked to key leukemogenic pathways, including Wnt/β-catenin and MAPK signaling, indicating it to be a potential target. Molecular docking of A. javanica compounds revealed CIDs 15559724, 5490003, and 74819331 as potent DDX3X inhibitors with strong binding affinity and favorable pharmacokinetic and toxicity profiles compared to RK-33. This study highlights the importance of DDX3X in LAML pathogenesis and suggests targeting it using plant-derived inhibitors, which may require further in vitro and in vivo validation. Full article

Background: Chronic insomnia (CID) is a highly prevalent sleep disorder, yet the precise mechanisms underlying it remain incompletely understood. The aim of this study is to analyze effective connectivity between key regions of the default mode network (DMN), executive control network (ECN), and salience network (SN) in patients with CID as potential neurologic correlates of the hyperarousal state. Methods: Thirty-one CID patients and 24 healthy controls (HC) were recruited. All the subjects filled out the Insomnia severity index scale (ISI), Beck depression inventory (BDI), and Epworth sleepiness scale (ESS), underwent polysomnography, and were scanned on functional magnetic resonance imaging. Statistical Parametric Mapping 12 was used to analyze the results. Spectral dynamic causal modeling was applied to the chosen regions of interest. Results: There were three significant connections present in the CID group—inhibitory from the dorsolateral prefrontal cortex (DLPFC) to the right hippocampus (Hippocamp R); excitatory from the dorsomedial prefrontal cortex to the ventromedial prefrontal cortex; and excitatory from the common medial prefrontal cortex to the right anterior insula (AIR). Two statistically significant excitatory connections were lacking in the patients’ group—from the posterior cingulate cortex (PCC) to AIR, and from precuneus to PCC. CID patients scored higher on the ISI and BDI. Significant negative correlations between DLPFC-Hippocamp R connectivity and both ISI and BDI scores were identified. Conclusions: Disruptions within the DMN and between the DMN, SN, and ECN reflect an impaired ability to appropriately shift between internally and externally directed cognitive states—an imbalance that potentially underlies the hyperarousal state of CID. Full article

Transformative digitally integrated pedagogy can enrich learning experiences, diversify the curriculum and broaden access to industry-relevant advanced clinical education for remote learners in medical education. Clinical skills are characterised as the portfolio of practical and interpersonal skills required by practicing clinicians. The purpose of this project was to design a new wholly online post-professional university subject for clinicians in different healthcare disciplines to advance these skills, which would traditionally be taught and assessed in-person. Our methodology included critically reviewing existing evidence of relevant medical skills which need to be included in the curriculum and approaches to their assessment. We designed a subject which dovetailed learning experiences with continuing clinical practice, and developed a new framework for remote video assessment of practical skills. Our pedagogical approaches included a backwards design coupled with a Four-Component Instructional Design Model (4C-ID) approach, which increased access and contextualised learning opportunities for diverse and practicing clinicians. Our narrative synthesis critically shares our experience and insights of embracing digital-technology opportunities while problem-solving to move past barriers. Our impact evaluation and experiential insights offer a platform to reimagine emerging possibilities for future digitally integrated education in medical education and other clinical-skills professions. Full article

Lysophosphatidylinositols are degradation products of phosphatidylinositols within cell membranes and digestive metabolites of a high-fat diet in the gut. G-protein-coupled receptor 55 (GPR55) is a receptor that senses lysophosphatidylinositol and acts as an immune mediator, being primarily upregulated during immune cell activation. This study aimed to investigate the role of GPR55, using its antagonist, CID16020046, in a collagen-induced rheumatoid arthritis mouse model. It was observed that DBA-1J mice develop joint lesions characteristic of rheumatoid arthritis following immunization with bovine type II collagen. The administration of CID16020046 (1 mg/kg, intraperitoneally) alleviated rheumatoid arthritis symptoms and inflammatory responses. Histopathological analysis showed that CID16020046 reduced foot edema, proteoglycan loss, and bone erosion in the joints. CID16020046 also decreased rheumatoid-arthritis-induced serum IgG levels, as measured using enzyme-linked immunosorbent assays. The treatment reduced levels of pro-inflammatory cytokines (IL-1β and IL-6), Th1 cytokine (IFN-γ), and Th17 cytokine (IL-17A), along with matrix metalloproteinase-3 (MMP-3) and the receptor activator of nuclear factor-κB ligand (RANKL) in the feet. A significant reduction in splenomegaly was also observed, along with significant reductions in CD4⁺ T helper 1 (Th1) and Th17 cells in the spleen. Additionally, CID16020046 suppressed the differentiation of naïve T cells into CD4⁺IL-17⁺ Th17 cells. CID16020046 suppressed expression levels of inflammatory cytokine mRNAs in SW982 human synovial cells. In conclusion, blocking GPR55 alleviates collagen-induced rheumatoid arthritis symptoms by suppressing Th1 and Th17 cells in the spleen and pro-inflammatory cytokines in the joints, suggesting that GPR55 is a potential therapeutic target for autoimmune inflammatory diseases. Full article