Search Results (625)

Bisphenol AF (BPAF) exposure is increasingly linked to metabolic disorders, yet the molecular initiating events (MIE) and key events (KE) leading to hepatic steatosis remain unclear. We constructed an adverse outcome pathway (AOP) to mechanistically connect BPAF-triggered macrophage–hepatocyte crosstalk to liver fat accumulation. Male C57BL/6 mice received daily oral gavage of 0, 0.5, 4, or 32 mg kg⁻¹ BPAF for 90 d, and Transwell co-cultures of RAW264.7 macrophages and AML12 hepatocytes were used for in vitro validation. Targeted metabolomics, western blotting, and lipid staining quantified succinate, pathway proteins, and steatosis. BPAF dose-dependently increased serum succinate (BMD = 6901.95 nM) and hepatic triglyceride (TG) (BMD = 874.26 nM). Cryo-EM docking revealed BPAF binding to SUCNR1 at 2.9 Å, disrupting the inactive-state conformation. In co-culture, BPAF-exposed macrophages released succinate that bound hepatocyte SUCNR1, suppressed Akt phosphorylation, and activated JNK. These KEs led to a 40% increase in lipid droplets and elevated TG, total cholesterol (TC), and free fatty acids (FFA) without liver weight gain. We propose the first AOP for BPAF-induced hepatic steatosis: BPAF–SUCNR1 binding (MIE) → macrophage succinate release (KE1) → SUCNR1-mediated Akt inhibition/JNK activation (KE2–4) → hepatic lipid accumulation (KE5) → steatosis (AO). These findings provide mechanistic insight for chemical risk assessment of BPAF and structurally related bisphenols. Full article

Cannabis use (CU) motives among older adults (OA) could be an important indicator of broader mental health. OA ages 60+ (N = 78) reported on CU, alcohol consumption, and mood and anxiety. Coping, enhancement, social, conformity, expansion, and routine motives were assessed. Relationships among CU, alcohol consumption, and screenings for Cannabis Use Disorder (CUD), Alcohol Use Disorder (AUD), depression, and anxiety were examined. OA who screened positive for CUD were not different in CU frequency or alcohol consumption, but did endorse higher routine, social, coping, and conformity motives than OA endorsing non-harmful CU (d = 1.01 to 1.70). Participants who screened positive for depression or anxiety endorsed higher coping (d = 1.87, 2.18) and routine (d = 0.83, 0.85) motives in the absence of higher alcohol or CU. Higher routine motives were particularly associated with positive CUD screening, beyond other motives and CU frequency. Healthcare providers serving OA with CU should ask about motives to help determine if further mental health evaluation is warranted. Full article

Alpha-synuclein (αsyn) misfolding and aggregation underlie several neurodegenerative disorders, including Parkinson’s disease. Early oligomeric intermediates are particularly toxic yet remain challenging to detect and characterize within cellular systems. Here, we employed the luminescent conjugated oligothiophene h-FTAA to investigate early aggregation events of human wildtype (huWT) and A53T-mutated αsyn (huA53T) both in vitro and in HEK293 cells stably expressing native human-αsyn. Comparative fibrillation assays revealed that h-FTAA detected αsyn aggregation with higher sensitivity and earlier onset than Thioflavin T, with the A53T variant displaying accelerated fibrillation. HEK293 cells stably expressing huWT- or huA53T-αsyn were exposed to respective pre-formed fibrils (PFFs), assessed via immunocytochemistry, h-FTAA staining, spectral emission profiling, and fluorescence lifetime imaging microscopy (FLIM). Notably, huA53T PFFs promoted earlier aggregation patterns and yielded narrower fluorescence lifetime distributions compared with huWT PFFs. Spectral imaging showed h-FTAA emission maxima (~550–580 nm) red-shifted and broadened in cells along with variable lifetimes (0.68–0.87 ns), indicating heterogeneous aggregate conformations influenced by cellular milieu. These findings demonstrate that h-FTAA is useful for distinguishing early αsyn conformers in living systems and, together with stable αsyn-expressing HEK293 cells, offers a platform for probing early αsyn morphotypes. Taken together, this opens for further discovery of biomarkers and drugs that can interfere with αsyn aggregation. Full article

Anxiety disorders represent a significant global health challenge, yet a substantial treatment gap persists, motivating the development of scalable digital health solutions. This study investigates the potential of integrating passive physiological data from consumer wearable devices with subjective self-reported surveys to predict state–trait anxiety. Leveraging the multi-modal, longitudinal LifeSnaps dataset, which captured “in the wild” data from 71 participants over four months, this research develops and evaluates a machine learning framework for this purpose. The methodology meticulously details a reproducible data curation pipeline, including participant-specific time zone harmonization, validated survey scoring, and comprehensive feature engineering from Fitbit Sense physiological data. A suite of machine learning models was trained to classify the presence of anxiety, defined by the State–Trait Anxiety Inventory (S-STAI). The CatBoost ensemble model achieved an accuracy of 77.6%, with high sensitivity (92.9%) but more modest specificity (48.9%). The positive predictive value (77.3%) and negative predictive value (78.6%) indicate balanced predictive utility across classes. The model obtained an F1-score of 84.3%, a Matthews correlation coefficient of 0.483, and an AUC of 0.709, suggesting good detection of anxious cases but more limited ability to correctly identify non-anxious cases. Post hoc explainability approaches (local and global) reveal that key predictors of state anxiety include measures of cardio-respiratory fitness (VO₂Max), calorie expenditure, duration of light activity, resting heart rate, thermal regulation and age. While additional sensitivity analysis and conformal prediction methods reveal that the size of the datasets contributes to overfitting, the features and the proposed approach is generally conducive for reasonable anxiety prediction. These findings underscore the use of machine learning and ubiquitous sensing modalities for a more holistic and accurate digital phenotyping of state anxiety. Full article

Background/Objectives: Intrinsically disordered protein regions (IDRs) are difficult to study due to their flexible nature and transient interactions. Computational folding using AlphaFold may offer one way to explore potential folding of these regions under various conditions. Human DNA topoisomerase IIα (TOP2A) is an essential enzyme involved in regulating DNA topology during replication and cell division. TOP2A has an IDR at the C-terminal domain (CTD) that has been shown to be important for regulating TOP2A function, but little is known about potential conformations that it may undertake. Methods: Utilizing the AlphaFold 3 (AF3) model by way of AlphaFold Server, TOP2A was folded as a dimer first without and then with 29 literature-supported post-translational modifications (PTMs) and DNA to observe whether there is predicted folding. Results: TOP2A CTD does not fold in the absence of PTMs. With the addition of PTMs, however, the CTD is predicted to fold into a globular bundle of loops and α-helices. While DNA alone did not induce folding, in the presence of PTMs, DNA ligands increased helicity of the folded CTD and caused it to interact at different core domain interfaces. In addition, DNA is predicted to enable folding of the TOP2A CTD in the presence of fewer PTMs when compared to the absence of DNA. Conclusions: AF3 predicts the folding of TOP2A CTD in the presence of specific PTMs, and this folding appears to shift to allow binding to DNA in functionally relevant regions. These studies provide predicted folding patterns that can be tested by biochemical approaches. AF3 may support the development of testable hypotheses regarding IDRs and enables researchers to model protein-DNA interactions. Full article

Abiotic stresses severely impair plant growth and productivity. To counteract stress, plants have evolved intricate strategies, including the induction of stress-responsive proteins. The Arabidopsis thaliana Salt Tolerance-Related Protein (STRP) has recently emerged as a key player in abiotic stress tolerance. STRP is a small, hydrophilic, intrinsically disordered protein that exhibits the potential to adopt distinct conformations depending on the cellular context. STRP is localized in the cytosol and nucleus and is associated with the plasma membrane. Stress induces the subcellular redistribution of STRP, accompanied by a significant increase (up to ten-fold) in its levels due to reduced degradation by the 26S proteasome. Reverse genetics studies have demonstrated that STRP can mitigate the detrimental effects of oxidative stress and participate in modulating stress-related gene expression. Although the exact mechanism of STRP remains unclear, its physicochemical properties suggest a dual role as a molecular shield, interacting with macromolecules without a fixed conformation, and as a binder of specific defense-related client proteins, adopting a defined tertiary structure. This review provides a comprehensive overview of STRP and its emerging role as a multifunctional player in abiotic stress responses, also highlighting its potential for strengthening crop resilience and maintaining agricultural productivity under global climate challenges. Full article

Diabetes mellitus is a characteristic metabolic disorder with diverse complications. α-Amylase and α-glucosidase, as key digestive enzymes regulating blood glucose, are important targets for diabetes prevention and management through their inhibition. This study investigated the inhibitory effects of six porphyrin compounds (TAPP, TCPP, THPP, Cu–TCPP, Fe–TCPP, Ni–TCPP) on two enzymes through in vitro inhibition assays, spectroscopic experiments, and molecular docking techniques. All six compounds effectively inhibited the activities of both enzymes. For α-amylase, the inhibitory potency (IC₅₀ = 13.03–245.04 μg/mL) followed the order TAPP > THPP > TCPP > Fe–TCPP > Ni–TCPP > Cu–TCPP. All six compounds exhibited more potent inhibitory activity against α-glucosidase (IC₅₀ = 0.24–25.43 μg/mL), with potency in the order of THPP > Ni–TCPP > Fe–TCPP > TCPP > Cu–TCPP > TAPP. Fluorescence quenching experiments revealed that all compounds statically quenched the intrinsic fluorescence of both enzymes (with Fe–TCPP exhibiting static-dominant mixed quenching against α-amylase), indicating complex formation. These interactions significantly altered the enzymes’ conformations, the microenvironments of Tyr/Trp residues, and secondary structure content, consequently reducing their catalytic activity. By examining the inhibitory impact of porphyrin compounds on α-amylase and α-glucosidase, this research establishes a vital experimental and theoretical basis for diabetes therapeutics. Full article

Orthoses are external devices designed to provide structural and functional support for disorders affecting the musculoskeletal or nervous systems. While these devices have a long history, recent technological advancements offer significant opportunities to enhance their therapeutic performance. This review examines three key innovations shaping the future of orthotic devices: additive manufacturing, auxetic metamaterials, and smart sensors. Additive manufacturing (AM), commonly known as 3D printing, is gaining prominence for its ability to create patient-specific solutions, improve design flexibility, and reduce production time. Despite these advantages, traditional fabrication methods remain dominant due to cost and regulatory challenges. Auxetic metamaterials, characterized by a negative Poisson’s ratio, allow an orthosis to dynamically conform to the patient’s anatomy and movements while maintaining stability and comfort. Thanks to synclastic deformation, auxetic structures reduce the formation of wrinkles during motion, improving body fit, and potentially enhancing comfort as well as adherence to orthosis usage recommendations. However, their integration into orthoses is still in the early stages, requiring further research and clinical validation. Finally, smart sensors have been extensively studied for the real-time monitoring of joint movement and rehabilitation progress, enabling personalized therapy and improved clinical outcomes. In conclusion, these emerging technologies—additive manufacturing, auxetic metamaterials, and smart sensors—hold great promise for next-generation orthotic devices, but widespread adoption will depend on addressing technical, economic, and practical limitations. Full article

Adverse childhood experiences (ACEs) are critical determinants of long-term psychological and physiological health outcomes. Transgender and gender nonconforming (TGNC) individuals are at increased risk for ACEs, including family rejection, peer victimization, and systemic discrimination. Despite the growing body of research on this topic, an updated synthesis of recent literature is needed to understand the evolving landscape of ACE-related risks and protective factors in TGNC populations. This systematic review, conducted according to PRISMA guidelines, examined studies published between 2020 and 2024 that were related to the association between ACEs and mental health outcomes in TGNC individuals. A comprehensive database search yielded 6124 articles, 42 of which met the inclusion criteria. Data extraction focused on the type of ACEs reported, associated mental health outcomes, resilience factors, and clinical implications. The results showed that TGNC individuals are significantly more likely to experience childhood maltreatment, including emotional, physical, and sexual abuse, as well as transphobia-specific ACEs such as forced gender conformity and identity denial. These experiences are associated with an increased risk of depression, PTSD, suicidality, and substance use disorders. Family rejection was found to be a critical risk factor, while social support, gender-specific care, and self-efficacy showed protective effects. The reviewed studies emphasize the urgent need for trauma-informed and gender-affirming mental health interventions to mitigate the effects of ACEs on TGNC individuals. Limitations and future research directions are discussed. Full article

Background: Biotinidase deficiency (BD) is a common autosomal recessive metabolic disorder in Qatar and the Arab world. It is treatable if detected early, making it essential to understand the genetic variants involved. This study aimed to investigate the carrier frequency of BD-related variants in a healthy Qatari population, reflecting the genetic landscape of the broader Middle Eastern region; classify them using bioinformatics tools; and compare findings with global datasets. Methods: Whole-genome sequencing data from 14,669 participants in the Qatar Genome Program (QGP), a multiethnic cohort including Qatari nationals and long-term residents (≥15 years), were analyzed to identify BTD variants. A total of 723, including 653 single-nucleotide polymorphisms (SNPs) and 70 structural variants (SVs) in BTD associated with BD, were screened against the Qatari cohort and compared with international data. In silico tools were used to assess variant pathogenicity, conservation, and protein stability. Molecular dynamics (MD) simulations were performed to evaluate structural and functional changes in the BTD. Results: A total of 80 SNPs and 3 SVs were identified, among which 21 variants (19 SNPs and 2 SVs) were classified as pathogenic or likely pathogenic, according to ClinVar. The carrier frequency of BTD-related variants in Qatar was 1:20, primarily driven by rs13078881 (D444H). Molecular dynamics (MD) simulations revealed significant conformational changes with H323R, D444H, and P497S, which demonstrated increased flexibility (higher RMSD/RMSF and PCA trace values). Additionally, R209C and D444H showed reduced compactness (higher Rg) and distinct energy minima, suggesting altered conformational states. Conclusions: This study demonstrates a high carrier frequency of pathogenic BTD variants in the Qatari population, underscoring the need to integrate these SNPs and SVs into the national genomic neonatal screening program (gNBS) for enhanced early detection and treatment strategies. The mild structural deviations observed in the D444H mutant through MD simulations may explain its association with milder clinical phenotypes of BTD, offering valuable insights for personalized therapeutic approaches. Full article

Gout and calcium pyrophosphate crystal deposition disease (CPPD) are frequently associated with comorbid disorders, including coronary artery disease and osteoarthritis, in which ectopic calcification with basic calcium phosphate crystals commonly affects arteries and articular cartilage, respectively. Accepting the 2024 G-CAN Gold Medal, I review my research philosophy for translational etiopathogenesis investigation in gout and CPPD, atherosclerosis, responses to arterial injury, and osteoarthritis. Since molecular homeostasis points to pathophysiology and vice versa, I have followed selected molecular players and pathways to phenotypes. Typically, behind each disease target is another target. Illuminating passageways between etiopathogenic pathways is especially productive when using approaches beyond conventional “omics” to reveal the impact of specific post-translational protein modifications, and changes in protein conformation, complex assembly, and interactomes. Highlighting these concepts, I review my past studies on specific molecular pathways, and current perspectives for the following: (i) PP_i, NPP1, ANKH, and transglutaminase 2 (TG2); (ii) relationships between NPP1, ANKH, Vanin-1 Pantetheinase, and ectopic chondrogenesis; (iii) intersections between adenosine, AMPK, CXCL8 and its receptor CXCR2, the receptor for advanced glycation endproducts (RAGE) and chondrocyte hypertrophy; (iv) lubricin homeostasis and proteolysis; (v) receptor for advanced glycation endproducts (RAGE) and TG2-catalyzed post-translational calgranulin modification; (vi) complement activation and C5b-9 assembly, and the nucleotide-bound conformation of TG2. The inescapable conclusion is that these molecular pathways tightly knit crystal arthropathy with both arterial and osteoarthritis comorbidity. Full article

Cryo-electron microscopy single-particle analysis (cryo-EM SPA) has advanced three-dimensional protein structure determination, yet resolving intrinsically disordered proteins and regions (IDPs/IDRs) remains challenging due to conformational heterogeneity. This research evaluates cryo-EM’s capacity to map dynamic regions, assesses the adaptability of disorder prediction tools, and explores optimization strategies for dynamic structure prediction. Cryo-EM SPA datasets from 2000 to 2024 were categorized into different periods, forming a database integrating sequence data and disorder indices. Established prediction tools—AlphaFold2 (pLDDT), flDPnn, and IUPred—were evaluated for transferability, while a multi-level CLTC hybrid model (combining CNN, LSTM, Transformer, and CRF architectures) was developed to link local conformational fluctuations with global sequence contexts. Analyses revealed consistent advancements in average resolution and model counts over the past decade, although mapping disordered regions remained technically demanding. Both the adapted AlphaFold pLDDT and the CLTC model demonstrated efficacy in predicting structurally variable and poorly resolved regions. A subset of the cryo-EM missing residues exhibited intermediate conformational features, suggesting classification ambiguities potentially influenced by experimental conditions. These findings systematically outline the evolving capabilities of cryo-EM in resolving dynamic regions, benchmark the adaptability of computational tools, and introduce a hybrid model to enhance prediction accuracy. This study provides a framework for addressing conformational heterogeneity, contributing to methodological advancements in structural biology. Full article

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterised by cognitive decline, synaptic loss, and multifaceted pathology involving amyloid-β (Aβ) aggregation, tau hyperphosphorylation, neuroinflammation, and impaired proteostasis. In recent years, biologic therapies, such as monoclonal antibodies, vaccines, antisense oligonucleotides (ASOs), and gene therapies, have gained prominence as promising disease-modifying strategies. In this review, we provide a comprehensive synthesis of current biologic approaches under clinical evaluation for AD. Drawing on data curated from ClinicalTrials.gov (as of 2025), we systematically summarise the molecular targets, therapeutic modalities, mechanisms of action, trial phases, and sponsors of over 60 biologic agents. These include Aβ-directed antibodies targeting distinct conformers such as protofibrils, pyroglutamate-modified species, and soluble oligomers; tau-targeted immunotherapies and RNA-based interventions; and emerging platforms focused on neuroimmune modulation, peptide hormones, and microbiota-based strategies. Gene and RNA therapeutics, particularly ASOs and small interfering RNAs (siRNAs) delivered intrathecally or via lipid nanoparticles, are also reviewed for their potential to modulate intracellular targets with high specificity. We also analyse the historical landscape of biologic candidates that failed to reach approval, discussing key reasons for trial discontinuation, including lack of clinical efficacy, safety concerns (e.g., amyloid-related imaging abnormalities), or inadequate biomarker responses. These cases offer crucial insights for refining future drug design. Looking ahead, we highlight major challenges and evolving perspectives in AD biologic therapy: expanding therapeutic targets beyond Aβ and tau, overcoming delivery barriers to the brain, designing prevention-oriented and genetically stratified trials, and navigating regulatory and ethical considerations. Together, these efforts signal a paradigm shift in AD drug development, from symptomatic treatment to mechanism-based precision biologics. By integrating real-time clinical trial data with mechanistic insight, this review aims to inform both translational research and therapeutic innovation in AD. Full article

Several carrier proteins are involved in nuclear translocation from the cytoplasm to the nucleus in eukaryotic cells. We have previously demonstrated the binding of several intact folded and disordered proteins to the human isoform importin α3 (Impα3); furthermore, disordered peptides, corresponding to their nuclear localization signals (NLSs), also interact with Impα3. These proteins and their isolated NLSs also bind to the truncated importin species ∆Impα3, which does not contain the N-terminal disordered importin binding domain (IBB). In this work, we added a further ‘layer’ of conformational disorder to our studies, testing whether the isolated D-enantiomers of NLSs of selected proteins, either folded or unfolded, were capable of binding to both Impα3 and ∆Impα3. The D-enantiomers, like their L-form counterparts, were monomeric and disordered in isolation, as shown by nuclear magnetic resonance (NMR). We measured the ability of such D-enantiomeric NLSs to interact with both importin species by using fluorescence, biolayer interferometry (BLI), isothermal titration calorimetry (ITC), and molecular simulations. In all cases, the binding affinities were within the same range as those measured for their L-isomer counterparts for either Impα3 or ∆Impα3, and the binding locations corresponded to the major NLS binding site of the protein. Thus, the stereoisomeric nature is not important in defining the binding of proteins to the main component of classical cellular translocation machinery, although the primary structure of the hot-spot site for NLS binding of importin is well defined. Full article

This study presents the development and the mechanical and clinical characterization of a flexible biodegradable chitosan–glycerol–graphite composite strain sensor for real-time respiratory monitoring, where the main material, chitosan, is derived and extracted from Tenebrio Molitor larvae shells. Chitosan was extracted using a sustainable, low-impact protocol and processed into a stretchable and flexible film through glycerol plasticization and graphite integration, forming a conductive biocomposite. The sensor, fabricated in a straight-line geometry to ensure uniform strain distribution and signal stability, was evaluated for its mechanical and electrical performance under cyclic loading. Results demonstrate linearity, repeatability, and responsiveness to strain variations in the stain sensor during mechanical characterization and performance, ranging from 1 to 15%, with minimal hysteresis and fast recovery times. The device reliably captured respiratory cycles during normal breathing across three different areas of measurement: the sternum, lower ribs, and diaphragm. The strain sensor also identified distinct breathing patterns, including eupnea, tachypnea, bradypnea, apnea, and Kussmaul respiration, showing the capability to sense respiratory cycles during pathological situations. Compared to conventional monitoring systems, the sensor offers superior skin conformity, better adhesion, comfort, and improved signal quality without the need for invasive procedures or complex instrumentation. Its low-cost, biocompatible design holds strong potential for wearable healthcare applications, particularly in continuous respiratory tracking, sleep disorder diagnostics, and home-based patient monitoring. Future work will focus on wireless integration, environmental durability, and clinical validation. Full article