Excessive alcohol consumption leads to neurodegeneration, driven primarily by oxidative stress and mitochondrial dysfunction, yet no specific treatment exists. Nicotinamide riboside chloride (NRC), a nicotinamide adenine dinucleotide precursor, has demonstrated therapeutic potential in mitigating mitochondrial dysfunction in heart failure, but its role in alcohol-induced neurodegeneration remains unexplored. This study investigated NRC’s neuroprotective effects using behavioral tests, serum ethanol and inflammatory marker analysis, hematoxylin-eosin staining, and molecular assays of in vitro models. Proteomics and GEO database analysis further elucidated the mechanisms of alcohol-induced brain injury. Results showed that NRC significantly improved alcohol-related cognitive impairment and neuroinflammation. Both our experimental data and external datasets identified mitochondrial dysfunction as a key driver of alcohol-induced neuronal damage, characterized by impaired mitophagy and disrupted mitochondrial unfolded protein response (UPRmt). NRC supplementation restored mitochondrial homeostasis by enhancing UPRmt and Fundc1-dependent mitophagy. Mechanistically, UPRmt inhibition abolished NRC’s protective effects by suppressing Fundc1 expression and mitophagy, whereas mitophagy inhibition did not affect UPRmt, suggesting a hierarchical regulation where UPRmt governs Fundc1-mediated mitophagy. In conclusion, alcohol disrupts mitochondrial quality control, but NRC counteracts neuronal toxicity by activating UPRmt and restoring Fundc1-driven mitophagy, offering a promising therapeutic strategy for alcohol-related neuronal damage.
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Urinary tract infections (UTIs) are one of the most common infectious diseases worldwide, predominantly caused by Escherichia coli. We constructed a reporter phage T4::Nluc to achieve rapid, sensitive, and specific detection of Escherichia coli in UTIs.
T4::Nluc was constructed using the CRISPR/Cas9 system combined with homologous recombination and was confirmed through Sanger sequencing. The biological properties of T4 and T4::Nluc were compared. Time‐luminescence curves were detected to investigate the limit of detection (LOD) and the influence of urine. Additionally, the specificity of T4::Nluc was examined by co‐culturing it with other pathogens. In total, 104 urinary Escherichia coli isolates were collected to assess detection coverage. Finally, 698 urine samples were collected for clinical validation.
T4::Nluc was confirmed to be correct. The one‐step growth curves of T4 and T4::Nluc were similar, but the optimal multiplicity of infection for T4 was 1, and that for T4::Nluc was 0.1, indicating that genetic modification had some effect. The LOD was 104 colony‐forming unit/mL detected at 220 min. Urine did not affect detection and T4::Nluc did not cross‐react with other pathogens. T4::Nluc could detect 38.46% of clinical strains, demonstrating higher sensitivity than the double‐layer overlay assay (25.96%). In clinical urine samples, its detection sensitivity was 36.59%, and the specificity was 100%.
T4::Nluc was successfully constructed and could detect Escherichia coli with superior sensitivity and specificity compared with traditional diagnostics, fulfilling the diagnostic criteria for UTIs while significantly reducing the detection time. This presented a novel approach for rapid and accurate detection of E. coli in UTIs.
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