Insulin secretion can be stimulated by immune and neuronal processes prior to a rise in blood glucose, exemplified by the cephalic phase of insulin response in the anticipation of food. Pancreatic α-cells prevent hypoglycemia by releasing glucagon. Here, we identified α-cells as critical mediators of IL-1β- and cholinergic agonist-driven insulin secretion. Cholinergic blockade prevented glucagon-stimulated insulin secretion in mice. Selective ablation of α-cells abolished cephalic phase insulin release. Islets from α-cell-deficient mice also failed to secrete insulin in response to IL-1β or muscarinic receptor activation. However, glucagon, acting on glucagon and GLP-1 receptors, rescued this insulin-stimulatory response. Mechanistically, intracellular Ca2+ mobilization at fasting glucose mediated cholinergic and IL-1β-stimulated insulin release. Short-term high-fat diet impaired glucagon-induced insulin secretion in vivo, while isolated islets showed increased insulin secretion after cholinergic stimulation versus chow-fed controls. These findings reveal α-cell-derived glucagon as a gatekeeper of immune and neuronal control of insulin secretion at fasting glucose.
Hippocampal sclerosis of aging (HS-A), a common cause of dementia, can currently only be diagnosed at autopsy. We aimed to identify and evaluate MRI metrics to distinguish HS-A from Alzheimer's Disease neuropathologic change (ADNC) and cases with limited/no pathology. HS-A (N = 5), ADNC (N = 10), and limited/no pathology (N = 12) cases were compared on postmortem MRI signatures: manually measured cornu ammonis (CA)1/subiculum thickness, grey matter signal intensity, and automated hippocampal subfield thickness metrics. Similar metrics were obtained in T1-weighted antemortem MRI in an initial dataset (HS-A = 4, ADNC = 7, limited/no pathology = 25) for group differences and discrimination (HS-A vs ADNC). T1-weighted metrics were then evaluated in a second dataset (HS-A = 6, ADNC = 18) and in a pooled post-hoc analysis combining HS-A and ADNC cases from both datasets (NHS-A = 10, NADNC = 25). Postmortem MRI showed hippocampal thinning and grey matter hypointensity in HS-A at the CA1-subiculum junction more severe than in ADNC and limited/no pathology cases. In antemortem MRI, differentiating HS-A and ADNC based on anterior/posterior manual measures of CA1/subiculum thickness and hippocampal volumes displayed good discrimination in dataset 1, but lower discriminative performance in dataset 2. In complementary analyses pooling both datasets and adjusting for age, manual thickness achieved good performance (area under the curve (AUC) = 0.80-0.87), while anterior, posterior, and whole hippocampal volumes showed excellent discrimination (AUC = 0.94-0.98). The study included a relatively small and neuropathologically heterogeneous sample. The final antemortem analyses were exploratory, reflecting challenges in replicating findings across independent datasets. Classification was limited to HS-A and ADNC, leaving it uncertain how the metrics perform when comparing HS-A with other diagnostic groups. HS-A diagnoses were determined postmortem, often several years after MRI, and most measures relied on standard imaging sequences with limited resolution to assess fine-grained hippocampal subfields. HS-A displays distinct changes in the hippocampus that are detectable through structural MRI. Associated quantifiable MRI metrics may serve as promising tools in aiding antemortem HS-A diagnosis but require further validation in larger cohorts and against other dementia-related diseases.
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DNA methylation, particularly 5-methylcytosine (5 mC), is a key epigenetic modification involved in the regulation of gene expression and genomic stability, and has emerged as a promising biomarker for early cancer screening and molecular stratification. CRISPR-Cas12a systems have been increasingly exploited to convert methylation-associated sequence information into detectable signals owing to their programmability and collateral cleavage-mediated signal amplification. However, many CRISPR-based assays remain constrained by the high background and relatively slow response kinetics of conventional fluorophore-quencher reporters. To overcome these limitations, we developed a cleavage-responsive DNA-templated silver nanocluster (DNA/AgNC) reporter that translates Cas12a trans-cleavage activity into a green-to-red ratiometric fluorescence shift. In this design, the AgNC-templating DNA scaffold itself serves as an enzymatically cleavable signal transducer, rather than relying on a terminal fluorophore-quencher pair. Compared with a representative F-ssDNA-Q reporter, the DNA/AgNC reporter exhibited stronger apparent association with Cas12a and an approximately two-fold improvement in apparent catalytic efficiency. When incorporated into an MSRE-RPA-Cas12a workflow, the platform achieved a limit of detection of 74.5 aM, while completing the Cas12a reporting step within 30 min. Coupling this assay with a miniaturized optoelectronic device further enabled spatially resolved profiling of five genomic loci, with relative errors of approximately 5%. Overall, this strategy establishes a ratiometric reporter format for CRISPR-based DNA methylation profiling and offers potential for point-of-care epigenetic biosensing.
Nanobody-based therapeutics that incorporate multifunctional drug molecules into nanobodies (nanobody-drug conjugates, NDCs) have gained substantial attention as promising therapeutic strategies in biomedicine due to their unique ability to precisely target specific cancer cells. However, the efficacy of NDCs depends on efficient internalization of the antibody-antigen complex to facilitate intracellular drug release in the lysosome, and not all surface antigens suffer from internalization upon antibody binding, posing a significant limitation to the effectiveness of NDCs. To address this gap, we rationally designed and constructed MT3 and anti-PD-L1 nanobody (Nb-PD-L1:KN035) fusion protein that enabled self-assembly into nanoparticles during the expression process. The MT3 domain of MT3-KN035 fusion protein nanoparticles did not interrupt the binding affinity between KN035 and PD-L1. In addition, MT3-KN035 protein nanoparticles not only migrated from the PD-L1-positive cell surface to intracellular lysosomes in vitro but also hold a huge targeting potential for tumor regions in vivo. Meanwhile, MT3-KN035 conjugated multiple aldoxorubicin (hereafter, these conjugates are referred to as MT3-KN035-DOX) via an acidic cleavable linker; MT3-KN035-DOXHigh induced tumor apoptosis and exhibited significant antitumor efficacy via facilitating immune cell infiltrations. This work provides a novel therapeutic strategy via nanobody-mediated target engagement and internalization that achieves synergistic therapeutic efficacy between chemotherapy and immune checkpoint blockade therapy.
The natural antifungal peptide Histatin 5 (Hst 5) is a histidine-rich cationic peptide secreted by human salivary glands and a key component of oral innate immunity, but its moderate activity limits clinical use. Hst 5 enters Candida albicans via the membrane receptor Ssa1/2. Here, we integrated artificial intelligence-assisted and computer-aided drug design to rationally modified the sequence structure of Hst 5. Truncated derivatives of Hst5 were screened for antimicrobial potential using ESM2-AFPpred, and high-probability candidates were docked with Ssa1/2. The Hst 5-22 was identified, then redesigned based on alanine scanning to yield the optimized derivative Hst 5-22-RW. Compared with Hst 5, Hst 5-22-RW has a shorter sequence, stronger Ssa1/2 binding, and improved activity against C. albicans. It also shows superior activity against fluconazole-resistant strains. RT-qPCR and transmembrane tracking confirmed higher cellular transport efficiency in C. albicans. The CADD/AIDD-driven optimization successfully generated the highly active antifungal peptide Hst 5-22-RW, providing a novel strategy for rational modification of antimicrobial peptides.
PocketMaster is a flexible and automated tool for the analysis, clustering, and interpretation of protein pockets, enabling the exploration of structural diversity in functional and interacting regions of proteins. The tool provides multiple strategies for defining pocket alignment regions, along with various alignment algorithms and clustering approaches, allowing analyses to be customized for different research objectives. In addition, it automatically documents results and provides informative visualizations and reports. With these capabilities, PocketMaster can be particularly valuable in the early stages of drug design, where accurate analysis and selection of protein structures are essential. Using TYK2 as a case study, PocketMaster demonstrates its ability to identify conformational differences between kinase and pseudokinase domains, as well as subtypes of structures within each domain, reflecting the influence of various ligands and protein states. The estrogen receptor alpha (ERα) ligand-binding pocket was also analyzed as an additional case study, showing the tool's performance in capturing conformational variations in helix 12 (H12) between active (agonist-bound) and inactive (antagonist-bound) states. The results confirm known structural features and illustrate the potential of the tool for systematic exploration of protein pockets, quantitative assessment of differences, and support of rational drug design. The PocketMaster source code, together with example input files and documentation, can be accessed at https://github.com/narek-abelyan/PocketMaster.
Antimicrobial resistance (AMR) has emerged as a major global health challenge, contributing to nearly 5 million deaths annually, according to recent WHO reports. Nanotechnology offers an alternative, with biologically produced nanoparticles being regarded as sustainable and potent antimicrobial agents. Iron oxide nanoparticles (IONPs), particularly Fe3O4 and Fe2O3, are of particular interest due to their unique magnetic, physicochemical, and bio-functional properties. The review discusses the reduction mechanism that explains the biological production of IONPs using microbial biomolecules and phytochemicals as natural capping, stabilizing, and reducing agents. Particular attention is given to biomolecules-mediated electron transfer, Fe2+ and Fe3+ redox cycling during the nucleation process and how surface functionalization determines nanoparticles' stability and activity. Additionally, this review discussed how these reduction mechanisms directly affect antimicrobial action through the generation of reactive oxygen species (ROS), membrane rupture, DNA/protein damage, and biofilm inhibition. A comparative analysis of plant and microbe-mediated synthesis is presented, along with structure-activity correlates that regulate antibacterial activity. Applications of biogenic IONPs are examined in the realms of biomedicine, industry, and the environment, with particular focus on their potential use in combination therapy to prevent antibiotic resistance. The positive factors of eco-friendliness, increased biocompatibility and multifunctionality are offset by such factors as variability in synthesis, scaling and long-term safety. Finally, in the future, the combination of clinical translation regulatory frameworks, omics-based mapping of bacterial responses, and molecular-level mechanistic investigations will be of relative importance. Altogether, biologically synthesized IONPs are an eco-friendly and effective approach to prevent microbial resistance that can be applied to the interface between green chemistry and more sophisticated nanomedicine. This review uniquely integrates comparative plant- and microbial-mediated synthesis, mechanistic Fe3+/Fe2+ reduction pathways, physicochemical optimization parameters, and molecular antimicrobial mechanisms involved in combating multidrug-resistant pathogens.
This study investigates the use of RADA16 hydrogels for delivering carnosic acid (CA) in glaucoma treatment. Rheological tests at 0.5% (w/v), 1% (w/v), and 2% (w/v) concentrations showed significant elasticity, with Young's moduli of 30.18 Pa, 16.22 Pa, and 26.66 Pa, respectively. The 1% (w/v) concentration had the lowest stiffness, ideal for intravitreal injection. SEM revealed the hydrogel's porous microstructure. The RADA16-CA system provided sustained CA release over 72 h, peaking at 6 h. In vitro, high CA concentrations (≥ 256 μg/mL) were cytotoxic, but RADA16 and RADA16-CA enhanced cell proliferation. RADA16-CA demonstrated superior efficacy and biocompatibility. In an acute ocular hypertension model, RADA16-CA improved retinal health in rats by reducing retinal nerve fiber layer thickness, ROS-positive cells, and pro-inflammatory cytokines, while boosting cell survival and antioxidant activity.These findings highlight the potential of the RADA16-CA sustained-release system as an optimized approach for glaucoma treatment.
Androgens act through the androgen receptor (AR), which regulates nearly a thousand genes. The human AR gene contains polymorphic repeats, including (CAG)n and (GGN)n, which affect AR transactivation. This study investigated their independent and combined effects on reproductive and general health. The study included 866 patients with male factor infertility (mean age: 32.8 years, and standard deviation: 6.8 years). Standard protocols were followed for semen analysis, phenotyping, and laboratory data collection. Repeat numbers of (CAG)n and (GGN)n polymorphisms were detected simultaneously using an established genotyping assay. Significantly lower sperm counts were observed in carriers of the AR gene with ≥24 compared to ≤22 GGN repeats (median: 13.5 × 106 vs 18.2 × 106 per ejaculate, P < 0.01). The meta-analysis with the Baltic young men cohort confirmed this association (n = 1843; linear regression: β = -0.38 × 106 [95% confidence interval, 95% CI: -0.75 × 106 to -0.01 × 106] per ejaculate, P = 0.044). The effect was further enhanced by long AR (CAG)n tract (≥25 repeats). The lowest sperm counts (median: 13.6 × 106 per ejaculate) and concentrations (3.5 × 106 ml-1) were detected in carriers of the AR haplotype combining ≥24 GGN and ≥25 CAG repeats (6.8% of patients). For AR (CAG)n repeats, a positive association was observed only with body mass index (BMI; P = 0.02). Neither AR repeat stretch affected semen volume, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, glucose, lipids, uric acid, or C-reactive protein (CRP) levels. In conclusion, an increased AR (GGN)n repeat number exerts a pronounced negative modulatory effect on sperm parameters. To date, only a limited number of common genetic variants have been reported to be associated with quantitative sperm parameters.
Intracavitary drug instillation is a crucial therapeutic strategy for treating bladder cancer. However, current methods are limited in efficacy due to insufficient tumour targeting and drug penetration across tissue barriers in pathophysiological conditions. Here we devise biohybrid magnetic algae microrobots with hierarchical nanoporous structure and develop an 'algebot'-mediated, non-contact convective transport strategy to synergistically integrate targeted carrier transport, selective drug release and ultrafast tissue penetration. Our approach leverages machine-intelligent image feedback for autonomous navigation, magnetite-endowed multimodal control for reconfigurable swarming and flow-tuned convective diffusion for on-demand therapeutic delivery. We exemplify this approach with doxorubicin-loaded magnetic Coscinodiscus granii evaluated in a murine model of bladder tumour, demonstrating an over tenfold increase in drug permeation and substantially reduced tumour burden to less than 3% compared with conventional intravesical instillation in a preclinical trial of 1-week therapy without inducing systemic toxicity. Our drug delivery system offers a non-invasive solution to overcome complex biological barriers, advancing the efficacy and safety of intracavitary chemotherapy.
Bone remodeling is orchestrated by the balanced activity of bone-resorbing osteoclasts and bone-forming osteoblasts. Interferon regulatory factor 5 (IRF5), a member of the IRF family of transcription factors, serves as a critical regulator of macrophage immune-related activity. However, macrophages also serve as the precursor cell of osteoclasts where the role of IRF5 in osteoclast-mediated bone remodeling in vivo remains undefined. Here, we find that IRF5 exhibits nuclear localization specifically in preosteoclasts during macrophage-osteoclast transition in vitro. In turn, myeloid cell-specific Irf5 conditional knockout (Irf5ΔM/ΔM) mice exhibit a significant osteopenic phenotype. Unexpectedly, osteoclast activity remained unaltered, while osteoblastic bone formation was significantly reduced. Interestingly, while conditioned media from wild-type preosteoclasts and osteoclasts increased the osteogenic potential of osteoblastic cells, this stimulatory activity was largely abrogated in conditioned media recovered from Irf5ΔM/ΔM preosteoclasts and osteoclasts. Further, the osteogenic potential of bone marrow stromal cells was markedly inhibited when co-cultured with Irf5ΔM/ΔM osteoclasts. Complementing these findings, genome wide analysis revealed that the Irf5-deficient osteoclast lineage displayed major changes in transcriptional programs related to extracellular matrix organization, bone development, and Notch signaling. Taken together, our study identifies IRF5 as a previously unrecognized transcriptional regulator of osteoblast-stimulating factors within the osteoclast lineage, thereby offering new insights on osteoimmune regulation of bone remodeling.
Do the genetic and clinical outcomes of monopronuclear blastocysts (MPBs) differ between standard insemination and intracytoplasmic sperm injection cycles, and what do these differences imply for risk stratification and individualized clinical decision-making? IVF-derived MPBs demonstrate significantly lower rates of uniparental inheritance than ICSI-derived MPBs (96.9% vs 65.9%) biparental inheritance (P < 0.001), with comparable clinical and neonatal outcomes to 2PN blastocysts following transfer of euploid biparental embryos. A proportion of monopronuclear (1PN) zygotes can develop into euploid blastocysts and, following transfer, result in healthy live births, yet these embryos are widely discarded following fertilization check due to atypical pronucleation. The mechanisms underlying 1PN formation are varied and include asynchronous pronuclear formation, early pronuclear fusion, and premature pronuclear breakdown, meaning a subset may represent normally fertilized diploid zygotes missed at static assessment. Retrospective cohort study of 1PN embryos (N = 13 203) derived from IVF (n = 5266) or ICSI (n = 5464) inseminations across 10 730 cycles performed at multiple Australian clinics between January 2010 and December 2023. Embryos were defined as 1PN by the appearance of a single pronucleus at fertilization check 16-18 h post-insemination. Time-lapse footage was reviewed on Day 3 to identify late appearing 1PNs and exclude late second pronucleus appearance. Suitable blastocysts underwent trophectoderm biopsy for pre-implantation genetic testing for aneuploidy (PGT-A) and short tandem repeat (STR)-based biparental inheritance testing; only euploid embryos with confirmed biparental inheritance were available for frozen embryo transfer. Outcomes assessed included ploidy, biparental inheritance, blastocyst development, utilization, morphokinetics, pregnancy, live birth, and maternal and neonatal outcomes. IVF-derived MPBs had similar aneuploidy rates to two pronuclei (2PN) embryos (37.3% vs 33.9%) and 440/454 (96.9%) demonstrated biparental inheritance. ICSI-derived MPBs had higher aneuploidy rates (45.5% vs 31.5%, P < 0.05) and only 108/164 (65.9%) had biparental inheritance. Uniparental inheritance was predominantly maternal (IVF 92.8%; ICSI 94.6%). Both IVF and ICSI MPBs were less likely to reach blastocyst stage by Day 5 than 2PN embryos (IVF 19.3% vs 63.3%; ICSI 9.7% vs 60.6%, P < 0.05), and biparental IVF-1PN zygotes were more likely to have more nucleoli compared with uniparental IVF-1PN zygotes (P = 0.008). For embryos with confirmed biparental inheritance, there was no significant difference in clinical pregnancy, ongoing pregnancy, live birth rates, or neonatal outcomes compared with 2PN blastocysts. In approximately one in six cycles containing a 1PN embryo, no utilizable 2PN embryo were available (IVF 15.6%; ICSI 16.7%), with the 1PN embryo representing the sole option for embryo utilization. Retrospective single-entity design introduces potential selection bias and limits generalizability. Uniform protocols across sites preclude the level of evidence required for formal guideline revision. Differential use of time-lapse imaging for ICSI versus static assessment for IVF embryos may contribute to differences in 1PN identification rates between fertilization methods. The STR-based biparental classification platform has not been validated against an orthogonal technology for parental origin calling in 1PN embryos, and the possibility of triploid misclassification or absorption into unreported inconclusive outcomes cannot be excluded. These findings support a risk-stratified approach to MPB management based on fertilization method. IVF-derived MPBs meeting specific morphological and developmental criteria demonstrate a low-risk profile that warrants reconsideration of genetic testing requirements and may inform individualized consent discussions, particularly where 2PN embryos are unavailable. ICSI-derived MPBs carry a substantially higher risk of uniparental inheritance and comprehensive genetic testing remains indicated. No funding was attached to this study. The authors declare no conflict of interest. N/A.
Liquid chromatographic (LC) and capillary electrokinetic chromatography (EKC) methods with ultraviolet/diode array detection were developed for the enantioseparation of five azoles: econazole (ECO), miconazole (MICO), imazalil (IMA), ketoconazole (KET), and penconazole (PEN). The chiral selectors investigated were human serum albumin (HSA) and eight cyclodextrins (CDs), exploited in LC and EKC, respectively. HSA LC offered partial enantioseparation of IMA and, less effectively, of PEN. Interestingly, these azoles share a high degree of molecular similarity, as confirmed by in silico calculations. This may suggest that a single enantiomer achieves preferential access to enantioselective sites of the protein. To address the limited enantioselectivity of HSA for these azoles, CDs were explored as chiral selectors where EKC was preferable to LC due to its superior separation efficiency. The chiral discrimination capabilities of eight CDs were evaluated for these azole derivatives, aiming for fast separations with minimal amounts of CDs. Effective enantioseparation was achieved for ECO, MICO, IMA, and KET, using two of the tested CDs: 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) and heptakis(2,3,6-tri-O-methyl)-β-cyclodextrin (TM-β-CD). Satisfactory resolutions were obtained using a 40 mM phosphate buffer (pH 3.2) with 1 mM HP-β-CD for ECO and MICO and 8 mM TM-β-CD for KET. For IMA, the optimal conditions consisted of a 20 mM phosphate buffer with 3 mM HP-β-CD. None of the tested CDs were effective for the enantiomeric separation of PEN; therefore, a dual CD system was developed for the first time for this compound, employing succinyl-β-CD (Succ-β-CD) in combination with HP-β-CD in a borax buffer under basic conditions (pH 9.4), enabling complete enantioseparation of PEN. The low concentration of CDs required under optimal conditions renders the developed methods highly cost-effective.
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Mfd, the canonical bacterial transcription-repair coupling factor, is a highly conserved ATP-dependent DNA translocase with a complex architecture undergoing major rearrangements during its functional cycle. These changes regulate its ATPase and motor activities and are tuned by Mfd interactions with DNA, RNA polymerase and the UvrA subunit of the nucleotide excision repair excinuclease, Uvr(A)BC. Due to its role in accelerating molecular evolution and the development of antibiotic resistance, Mfd is also rapidly emerging as a prime target for the development of anti-evolution drugs to be administered in combination with narrow-spectrum antibiotics to prevent the rise of resistance and combat infection over a wider time window. Here we present the crystal structure of Thermus thermophilus Mfd in its nucleotide-free state. We note the pronounced disorder of the N-terminal UvrB homology module, which was previously seen in Escherichia coli to be engaged in a "clamp" interaction with the C-terminal domain, resulting in autoinhibition of its ATP-dependent functions. Thus, we conclude that the autoinhibitory interdomain interactions, such as the clamp, are not a universal feature of transcription-repair coupling factors. Consistent with this, Thermus thermophilus Mfd, unlike E. coli Mfd, translocates robustly on DNA even in the absence of RNA polymerase and displays DNA binding that is largely nucleotide independent. Our work brings mechanistic insight into the species-specific differences in Mfd structure and function and provides a structural framework for the design of anti-evolution drugs to combat antimicrobial resistance.
The micro/nanoplastics (MNPs) have been evidenced to exert detrimental effects on the blood-brain barrier (BBB) and the central nervous system (CNS). However, there is still a lack of effective research models on the mechanism of nerve injury caused by microplastics particles. This study focuses on analyzing the particle size characteristics of MNPs precipitated from plastic water bottles under different conditions of storage and uses 3D BBB microfluidic chips to assess the permeability and dynamic neurotoxicity of MNPs. The results showed that there was a significant increase in the average diameter of MNPs in purified water stored in plastic bottles. Moreover, the cultivation of BBB cells or neuronal cells with two different particle sizes of MNPs showed a significant decrease in cell survival rates. When MNPs were infused into the peripheral unit of the biomimetic chip, they could penetrate from the endothelial cell unit to the neuronal unit and induce a dynamic injury process with neuroinflammation, accompanied by tight junction disruptions, increased ROS levels, decreased mitochondrial membrane potential, decreased lipid droplet levels, and increased inflammatory effects. The research results based on engineering 3D microfluidic chips lay the foundation for a deeper understanding of the inflammatory damage to nerve cells caused by MNPs crossing the BBB.
Odorants in the inhaled air bind to odorant receptors embedded in cilia of olfactory receptor neurons, triggering the opening of the two ciliary olfactory transduction channels; first, a cationic heterotetrameric cAMP-gated channel, followed by an excitatory Ca2+-activated Cl- channel TMEM16B (also called anoctamin 2), which gives rise to a transduction current. While cholesterol is known to modulate ion channels, its effect on the olfactory channels, specifically TMEM16B, is unclear. We addressed how membrane cholesterol regulates these transduction channels. We heterologously expressed the main subunit of the olfactory cyclic nucleotide-gated (CNG) channel, CNGA2, and the Cl- channel TMEM16B and recorded their function in control, cholesterol-depleted, and cholesterol-enriched membranes. Maximal cAMP- or Ca2+-evoked currents were not altered by varying cholesterol levels. TMEM16B showed a progressive reduction in current upon repeated exposure to Ca2+ ("rundown"), which was accelerated in cholesterol-depleted membranes but decelerated in cholesterol-enriched patches when compared with control patches. Maximal cAMP-gated currents were stable over time, but their sensitivity was altered, with lower and higher cholesterol levels increasing and decreasing the channel's sensitivity, respectively. Cholesterol depletion of patches excised from mouse olfactory cilia containing native channels entirely abolished the TMEM16B current, while the maximal current of the native CNG channel remained unaltered. Reduced cholesterol did not change the sensitivity of the native CNG channel. Thus, while both olfactory transduction channels are sensitive to changes in membrane cholesterol in differing ways, the precise channel stoichiometry and the specific membrane environment also contribute to their function.
Social determinants of health (SDOH) may improve Alzheimer's disease (AD) risk prediction by capturing upstream contextual risk beyond routinely measured clinical variables. We aimed to develop and validate an accurate, interpretable machine-learning pipeline for AD risk prediction in UK Biobank using routinely collected data. Using data from 13 076 participants in the UK Biobank, we developed an automated machine-learning pipeline for AD risk prediction with feature selection and a C5.0 boosted-tree classifier. Data were split into training, development and test sets (7:2:1); missing values were imputed in the training data only, and feature selection, tuning and threshold calibration were performed using the training/development data, with final evaluation on the independent test set. Internal validation used repeated subsampling without replacement. During up to 16 years of follow-up, 927 participants developed AD. Feature selection reduced 3590 variables to 26 predictors spanning age, APOE4, SDOH, medical history and routine clinical measures. The final model showed good discrimination (area under the precision-recall curve 0.89) and adequate calibration (Hosmer-Lemeshow p=0.71), with stable performance under repeated subsampling. Sex-stratified models showed similar patterns. SDOH contributed useful predictive information, but their associations should be interpreted as predictive rather than causal and may reflect socioeconomic confounding and healthcare access. This model could support scalable AD risk screening using routinely collected data, but external validation and recalibration in non-UK populations are needed before broader application.
Rheumatoid arthritis (RA) is a chronic autoimmune disease driven by dysregulated Th17 cell responses and osteoclastogenesis. While metformin has shown potential in autoimmune modulation, its efficacy is often limited by high dosage requirements. In this study, we evaluated the therapeutic efficacy and underlying molecular mechanisms of SD282, a newly synthesized biguanide derivative, in treating RA. The anti-arthritic effects of SD282 were assessed using a collagen-induced arthritis (CIA) mouse model. Murine and human immune cells were used to investigate the impact of SD282 on Th17/Treg differentiation, STAT3 signaling, mitochondrial respiration (OCR), and osteoclastogenesis. SD282 treatment significantly ameliorated the clinical severity of CIA, reducing joint inflammation, cartilage damage, and bone erosion. At the cellular level, SD282 effectively inhibited Th17 differentiation and IL-17 production while promoting Foxp3 + Treg cells, thereby restoring immune homeostasis. Mechanistically, SD282 blocked STAT3 phosphorylation and normalized mitochondrial metabolic activity by enhancing OXPHOS complex potential. Furthermore, SD282 suppressed osteoclast differentiation by downregulating RANKL, MMP9, and NFATC1 signaling pathways. Our findings demonstrate that SD282 acts as a potent multi-target agent that suppresses RA progression by synergistically regulating the STAT3-Th17-Mitochondria axis and bone metabolism. SD282 represents a promising pharmacological candidate for the treatment of autoimmune arthritis with a superior mechanistic profile compared to conventional biguanides.