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The type V-F CRISPR-Cas12f system is a strong candidate for therapeutic applications due to the compact size of the Cas12f proteins. In this work, we identify six uncharacterized Cas12f1 proteins with nuclease activity in mammalian cells from assembled bacterial genomes. Among them, OsCas12f1 (433 aa) from Oscillibacter sp. and RhCas12f1 (415 aa) from Ruminiclostridium herbifermentans , which respectively target 5’ T-rich Protospacer Adjacent Motifs (PAMs) and 5’ C-rich PAMs, show the highest editing activity. Through protein and sgRNA engineering, we generate enhanced OsCas12f1 (enOsCas12f1) and enRhCas12f1 variants, with 5’-TTN and 5’-CCD (D = not C) PAMs respectively, exhibiting much higher editing efficiency and broader PAMs, compared with the engineered variant Un1Cas12f1 (Un1Cas12f1_ge4.1). Furthermore, by fusing the destabilized domain with enOsCas12f1, we generate inducible-enOsCas12f1 and demonstate its activity in vivo by single adeno-associated virus delivery. Finally, dead enOsCas12f1-based epigenetic editing and gene activation can also be achieved in mammalian cells. This study thus provides compact gene editing tools for basic research with remarkable promise for therapeutic applications. Cas12f proteins are small and sought after for therapeutic applications. Here the authors report six bacterial Cas12f1 proteins with nuclease activity in mammalian cells, perform sgRNA and protein engineering to generate variants with enhanced editing and broader PAMs, apply an inducible version in vivo.

This study aimed to compare the therapeutic effects of fresh (CMF) and dried (CMD) Centipeda minima against allergic rhinitis (AR), elucidate their underlying molecular mechanisms, and identify the bioactive compounds responsible for their immunomodulatory actions. An ovalbumin-induced AR mouse model was treated with CMF or CMD extracts, followed by evaluation of nasal symptoms, serum biomarkers (IgE, histamine, cytokines), and nasal mucosa histopathology. Transcriptomics and widely targeted metabolomics were integrated with network pharmacology to identify differentially expressed genes and bioactive components, which were further validated in RAW264.7 and RBL-2H3 cells. CMF and CMD exhibited distinct anti-AR mechanisms: CMF predominantly suppressed Th2 responses (reducing IgE, IL-6, and histamine while elevating IL-10), whereas CMD enhanced Th1 activity (increasing IFN-γ). Metabolomic analysis revealed CMF was rich in amino acids while CMD contained higher flavonoids, with neocnidilide and 6-gingerol identified as key bioactive compounds that modulated TNF-α, IL-6, and IL-10 via NF-κB and JAK-STAT pathways. These findings demonstrate that CMF and CMD exert complementary anti-inflammatory effects through Th2 inhibition and Th1 activation, respectively, providing a molecular basis for the traditional use of Centipeda minima and highlighting its bioactive compounds as potential therapeutics for inflammatory diseases.

Background Over recent decades, cutaneous melanoma (CM) incidence in the United States (US) has increased but now appears to be plateauing, whereas mortality has declined. However, much less is known about how CM incidence and mortality vary between states, or about how differences between states in incidence relate to lifestyle factors, ambient UV exposure, and prevention or health care resources. At the population level, most studies have compared overall CM incidence and mortality across broad racial and ethnic groups without describing age-specific or state-specific patterns. This lack of granularity limits our understanding of when and where different population groups bear the greatest CM burden and where targeted prevention and early detection efforts are most needed. Methods We conducted a cross-sectional study of the incidence and mortality trends of CM in the US from 2001 to 2019, including state-level analyses of temporal trends and ecological correlates, as well as national population-level analyses by age, sex, and race/ethnicity. Annual percent change (APC) and average annual percent change (AAPC) were calculated, and ecological correlations and multivariable linear regression models were fitted with state-level AAPC in CM incidence as the outcome. Results Between 2001 and 2016, CM incidence increased significantly (APC, 1.95% [95% CI, 1.67% to 2.24%], p  < 0.001) and then stabilized from 2016 to 2019 (APC, -0.59% [95% CI, -3.36% to 2.27%], p  = 0.662). During the study period, mortality declined significantly (AAPC, -1.52% [95% CI, -2.38% to -0.66%], p  < 0.001). A persistent upward incidence trend was observed in 24 states, predominantly in the Midwest and Southern regions. The incidence rate ratio (IRR) between the highest- and lowest-rate states increased from 2.63 in 2001 to 4.85 in 2019, whereas the mortality rate ratio (MRR) narrowed to 1.19 by 2019. In ecological analyses, higher state-level obesity prevalence and lower physical activity level were each associated with higher AAPC in CM incidence, whereas higher personal doctor rates were inversely associated. Average daily solar insolation and tanning/sunscreen policies were not significantly related to the AAPC. Across all racial/ethnic groups, CM incidence was higher in young women than in young men, with sex-specific crossover ages varying by race/ethnicity, while young men consistently experienced higher mortality. Age-stratified analyses showed that disparities in CM incidence and mortality rates between non-Hispanic Whites and other races/ethnicities were more pronounced in young individuals than in the elderly. There was also substantial interstate variation in incidence rate ratios comparing non-Hispanic Whites with other racial/ethnic groups. Conclusions Despite overall plateauing incidence and declining mortality, CM remains highly unequal across states and population groups. Interstate incidence disparities widened over time, with marked variation by age, sex, and race/ethnicity. These findings provide a more detailed picture of geographic and population-level disparities in CM and may help inform future surveillance, risk stratification, and hypothesis-driven prevention research.

Esophageal adenocarcinoma (EAC) has increased substantially to become the most common type of esophageal cancer in the United States, surpassing esophageal squamous cell carcinoma (ESCC). Whether the increasing incidence of EAC is linked to trends in esophageal cancer mortality is unclear. We analyzed esophageal cancer data from the Surveillance, Epidemiology, and End Results-12 cancer registry program, using Joinpoint regression for trend analysis and a decomposition method to attribute changes to population growth, population aging, and epidemiological changes. Age–period–cohort models were employed to estimate incidence and mortality through 2044. Between 1992 and 2019, 39,700 individuals were diagnosed with esophageal cancer, and 35,259 deaths were recorded from 1993 to 2019. The overall incidence of esophageal cancer declined [average annual percent change (AAPC), − 0.7%], but the incidence of EAC increased by an AAPC of 1.6% per year ( P  < 0.001) from 1992 to 2019. The overall mortality rates decreased by an annual percentage change of 1.0% [95% confidence intervals − 1.2% to − 0.7%] from 1998 to 2019, primarily related to decline in ESCC in 1996–2019. The mortality of EAC increased by an AAPC of 2.2% per year ( P  < 0.001) over the study period and increased for all demographic characteristic groups. Population aging and growth largely explain the increase in esophageal cancer over the last 3 decades. Future projections (2019–2044) suggest a 31% increase in incidence and deaths, with EAC rates continuing to rise (AAPC, 0.25; 95% CI 0.23–0.28). The number of esophageal cancer cases and deaths have significantly increased and projections indicate that this trend will continue. Effective measures must be taken to address the burden of esophageal cancer.

Duchenne muscular dystrophy (DMD) affecting 1 in 3500–5000 live male newborns is the frequently fatal genetic disease resulted from various mutations in DMD gene encoding dystrophin protein. About 70% of DMD-causing mutations are exon deletion leading to frameshift of open reading frame and dystrophin deficiency. To facilitate translating human DMD-targeting CRISPR therapeutics into patients, we herein establish a genetically humanized mouse model of DMD by replacing exon 50 and 51 of mouse Dmd gene with human exon 50 sequence. This humanized mouse model recapitulats patient’s DMD phenotypes of dystrophin deficiency and muscle dysfunction. Furthermore, we target splicing sites in human exon 50 with adenine base editor to induce exon skipping and robustly restored dystrophin expression in heart, tibialis anterior and diaphragm muscles. Importantly, systemic delivery of base editor via adeno-associated virus in the humanized male mouse model improves the muscle function of DMD mice to the similar level of wildtype ones, indicating the therapeutic efficacy of base editing strategy in treating most of DMD types with exon deletion or point mutations via exon-skipping induction. Duchenne muscular dystrophy (DMD) results from mutations in the DMD. Here the authors report using adenine base editing of exon splice sites to restore dystrophin expression and muscle function in a humanized mouse model of DMD with exon 50 and 51 of mouse DMD gene replaced with human exon 50.

Approximately 10% of monogenic diseases are caused by nonsense point mutations that generate premature termination codons (PTCs), resulting in a truncated protein and nonsense-mediated decay of the mutant mRNAs. Here, we demonstrate a mini-dCas13X-mediated RNA adenine base editing (mxABE) strategy to treat nonsense mutation-related monogenic diseases via A-to-G editing in a genetically humanized mouse model of Duchenne muscular dystrophy (DMD). Initially, we identified a nonsense point mutation (c.4174C>T, p.Gln1392*) in the DMD gene of a patient and validated its pathogenicity in humanized mice. In this model, mxABE packaged in a single adeno-associated virus (AAV) reached A-to-G editing rates up to 84% in vivo, at least 20-fold greater than rates reported in previous studies using other RNA editing modalities. Furthermore, mxABE restored robust expression of dystrophin protein to over 50% of WT levels by enabling PTC read-through in multiple muscle tissues. Importantly, systemic delivery of mxABE by AAV also rescued dystrophin expression to averages of 37%, 6%, and 54% of WT levels in the diaphragm, tibialis anterior, and heart muscle, respectively, as well as rescued muscle function. Our data strongly suggest that mxABE-based strategies may be a viable new treatment modality for DMD and other monogenic diseases.

Watershed nutrient balance studies traditionally focus on annual fluxes. In areas with strongly seasonal, Mediterranean-type climate regimes, riverine nutrient export may be greater during wet seasons when hydrologic forcing overwhelms or bypasses retention mechanisms. By combining data on riverine export with spatially detailed nutrient inputs, we examine how nitrogen (N) supply, retention, and streamflow shape annual and seasonal riverine N export in Oregon’s Willamette River Basin (WRB). The WRB has pronounced dry summers and wet winters, and the distribution of farmland, cities and forests create significant spatial variations in N inputs. Local data on N inputs were coupled with streamflow and chemistry to calculate fractional N export for 22 WRB sub-watersheds in the mid-2000s. For the entire WRB, 78% of the N inputs came from agricultural activities, mainly as synthetic fertilizer (69%); the next largest inputs were deposition (10%), alder fixation (5%) and point sources (5%). Crop-specific estimates of fertilizer agreed with county fertilizer sales rates at the high end of extension recommendations. Fractional riverine N export (annual riverine N export / net watershed N inputs) averaged 38% of net inputs in WRB tributaries, greater than other regions of North America. Fall and winter together accounted for 60–90% of the riverine N export across all watersheds. Summer export was small but was greatest in the watersheds that receive seasonal snowmelt. Large wet season losses, when biotic sinks are less active, result in a relatively high proportion of N inputs exported in this region with a Mediterranean climate and high runoff.

Magnesium (Mg) and its alloys have shown attractive biocompatibility and mechanical strength for medical applications, but low corrosion resistance of Mg in physiological environment limits its broad clinical translation. Hydroxyapatite (HA) nanoparticles (nHA) are promising coating materials for decreasing degradation rates and prolonging mechanical strength of Mg-based implants while enhancing bone healing due to their osteoconductivity and osteoinductivity. Conformal HA coatings with nano-to-submicron structures, namely nHA and mHA coatings, were deposited successfully on Mg plates and rods using a transonic particle acceleration (TPA) process under two different conditions, characterized, and investigated for their effects on Mg degradation in vitro . The nHA and mHA coatings enhanced corrosion resistance of Mg and retained 86–90% of ultimate compressive strength after in vitro immersion in rSBF for 6 weeks, much greater than non-coated Mg that only retained 66% of strength. Mg-based rods with or without coatings showed slower degradation than the respective Mg-based plates in rSBF after 6 weeks, likely because of the greater surface-to-volume ratio of Mg plates than Mg rods. This indicates that Mg-based plate and screw devices may undergo different degradation even when they have the same coatings and are implanted at the same or similar anatomical locations. Therefore, in addition to locations of implantation, the geometry, dimension, surface area, volume, and mass of Mg-based implants and devices should be carefully considered in their design and processing to ensure that they not only provide adequate structural and mechanical stability for bone fixation, but also support the functions of bone cells, as clinically required for craniomaxillofacial (CMF) and orthopedic implants. When the nHA and mHA coated Mg and non-coated Mg plates were cultured with bone marrow derived mesenchymal stem cells (BMSCs) using the in vitro direct culture method, greater cell adhesion densities were observed under indirect contact conditions than that under direct contact conditions for the nHA and mHA coated Mg. In comparison with non-coated Mg, the nHA and mHA coated Mg reduced BMSC adhesion densities directly on the surface, but increased the average BMSC adhesion densities under indirect contact. Further long-term studies in vitro and in vivo are necessary to elucidate the effects of nHA and mHA coatings on cell functions and tissue healing.

Endoplasmic reticulum stress (ER stress) contributes to the development of pulmonary fibrosis, especially in type II alveolar epithelial cells (AECs) apoptosis. ER stress also promotes NLRP3 inflammasome activation which is inhibited by upregulation of cAMP/PKA pathway. However, it is confused whether ER stress-induced NLRP3 inflammasome activation and pyroptosis in type II alveolar epithelial cells which exacerbates pulmonary fibrosis via a mechanism that is suppressed by cAMP/PKA pathway. In our research, we explored that potential links among NLRP3 inflammasome, ER stress, and cAMP/PKA pathway in type II AECs to explain the new mechanisms of pulmonary fibrosis. We found that in vivo, ER stress, NLRP3 inflammasome, and PKA upregulated in the alveolar epithelial area in animal models of pulmonary fibrosis. In addition, immunofluorescence staining further confirmed that ER stress, NLRP3 inflammasome, and cAMP/PKA had potential links on type II AECs in BLM group. In vitro, ER stress stimulated NLRP3 inflammasome activation, promoted pyroptosis, and also upregulated cAMP/PKA pathway. Upregulation of cAMP/PKA pathway inhibited ER stress-induced pyroptosis of A549 cells and vice versa. These results initially supported conclusion that ER stress may stimulate NLRP3 inflammasome activation and pyroptosis in type II AECs, which exacerbated pulmonary fibrosis, and cAMP/PKA pathway may act as a feedback regulator.

, a traditional Chinese medicinal herb, possesses cinnamaldehyde (CIN) with well-documented antifungal and immunomodulatory properties. Although CIN inhibits ( ) growth, its role in macrophage-mediated clearance remains poorly understood. Here, we evaluated CIN's antifungal activity using MIC determination, spot assays, and time-growth curves. Cell wall disruption (β-glucan and chitin exposure) was assessed by transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), and flow cytometry. Transcriptomic and functional enrichment analyses revealed that CIN compromises cell wall integrity by altering 123 differentially expressed genes (DEGs), particularly those governing hyphal development, cell wall biosynthesis, and biofilm formation. Specifically, CIN downregulated genes associated with β-glucan exposure, mannosylation, and chitin synthesis, and upregulated components of the Cek1/MAPK pathway. CIN-enhanced macrophage phagocytosis significantly increased fungal clearance and reduced fungal escape, as shown by flow cytometry, propidium iodide staining, and lactate dehydrogenase release assays. CIN-pretreated fungi activated the Dectin-1/Syk/CARD9/NF-κB cascade, leading to elevated pro-inflammatory cytokine secretion. Mechanistically, CIN induces β-1,3-glucan exposure on , thereby promoting Dectin-1-mediated phagocytosis and clearance. These findings provide an experimental basis for developing CIN as a novel antifungal therapeutic.