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Summary: The chaperonin CCT mediates folding of many cytosolic proteins, including G protein β subunits (Gβs). Here, we present a protocol for isolating Gβ5 bound to CCT and its co-chaperone PhLP1 and determining the CCT-mediated folding trajectory of Gβ5 using single-particle cryoelectron microscopy (cryo-EM) techniques. We describe steps for purifying CCT-Gβ5-PhLP1 from human cells, stabilizing the closed CCT conformation, preparing and imaging cryo-EM specimens, and processing data to recover multiple Gβ5 folding intermediates. This protocol permits visualization of protein folding by CCT.For complete details on the use and execution of this protocol, please refer to Sass et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Background: Although there have been several studies describing risk factors for complications after outpatient total hip arthroplasty (THA), data describing the timing of such complications is lacking. Methods: Patients who underwent outpatient or inpatient primary THA were identified in the 2012-2019 National Surgical Quality Improvement Program database. For 9 different 30-day complications, the median postoperative day of diagnosis was determined. Multivariable regressions were used to compare the risk of each complication between outpatient vs inpatient groups. Multivariable Cox proportional hazards modeling was used to evaluate the differences in the timing of each adverse event between the groups. Results: After outpatient THA, the median day of diagnosis for readmission was 12.5 (interquartile range 5-22), surgical site infection 15 (2-21), urinary tract infection 13.5 (6-19.5), deep vein thrombosis 13 (8-21), myocardial infarction 4.5 (1-7), pulmonary embolism 15 (8-25), sepsis 16 (9-26), stroke 2 (0-7), and pneumonia 6.5 (3-10). On multivariable regressions, outpatients had a lower relative risk (RR) of readmission (RR = 0.73), surgical site infection (RR = 0.72), and pneumonia (RR = 0.1), all P < .05. On multivariable cox proportional hazards modeling, there were no statistically significant differences in the timing of each complication between outpatient vs inpatient procedures (P > .05). Conclusions: The timing of complications after outpatient THA was similar to inpatient procedures. Consideration should be given to lowering thresholds for diagnostic testing after outpatient THA for each complication during the at-risk time periods identified here. Although extremely rare, this is especially important for catastrophic adverse events, which tend to occur early after discharge.

Introduction: Photodynamic therapy (PDT) is a safe, non-mutagenic, and non-scarring treatment for actinic keratoses (AK). Background: ‘Painless’ photodynamic therapy (p-PDT) is a regimen for AK that employs simultaneous aminolevulinate incubation and blue light illumination. The efficacy of p-PDT resembles that of traditional PDT, but detailed mechanisms of action for p-PDT are not well understood. Methods: To characterize the inflammatory effects of the p-PDT procedure 48 h following treatment and determine the association of inflammation with precancer burden, we performed a retrospective cohort study of 104 patients with AK of face or scalp treated with p-PDT between 2017 and 2019. Patients self-reported their side effects 48 h following p-PDT and took photographs of their face and scalp. Photographs were edited to define seven anatomic regions, and erythema was scored by four investigators. Results: Ninety-eight patients provided photographs suitable for erythema evaluation. Most patients experienced 2 or more side effects and some pain 48 h post-procedure. Females experienced more pain (p = 0.01) and side effects (p = 0.002) compared to males. AK burden was positively associated with post p-PDT erythema response (p < 0.0001) at all sites, but particularly in the temples (p = 0.002) and supralabial area (p = 0.009). Discussion: This study confirms a strong clinical inflammatory response after p-PDT. Severity of inflammation is positively associated with AK tumor burden, suggesting that post-treatment inflammation may be a pre-requisite for p-PDT efficacy. Interestingly, the results also identify certain gender-related differences in the severity of side effects experienced by patients post-PDT.

Summary: Uropathogenic E. coli (UPEC) is a primary organism responsible for urinary tract infections and a common cause of sepsis. Microbially experienced laboratory mice, generated by cohousing with pet store mice, exhibit increased morbidity and mortality to polymicrobial sepsis or lipopolysaccharide challenge. By contrast, cohoused mice display significant resistance, compared with specific pathogen-free mice, to a monomicrobial sepsis model using UPEC. CD115+ monocytes mediate protection in the cohoused mice, as depletion of these cells leads to increased mortality and UPEC pathogen burden. Further study of the cohoused mice reveals increased TNF-α production by monocytes, a skewing toward Ly6ChiCD115+ “classical” monocytes, and enhanced egress of Ly6ChiCD115+ monocytes from the bone marrow. Analysis of cohoused bone marrow also finds increased frequency and number of myeloid multipotent progenitor cells. These results show that a history of microbial exposure impacts innate immunity in mice, which can have important implications for the preclinical study of sepsis.

Alcohol use disorder (AUD) is characterized by loss of intake control, increased anxiety, and susceptibility to relapse inducing stressors. Both astrocytes and neurons contribute to behavioral and hormonal consequences of chronic intermittent ethanol (CIE) exposure in animal models. Details on how CIE disrupts hypothalamic neuro-glial communication, which mediates stress responses are lacking. We conducted a behavioral battery (grooming, open field, reactivity to a single, uncued foot-shock, intermittent-access two-bottle choice ethanol drinking) followed by Ca2+ imaging in ex-vivo slices of paraventricular nucleus of the hypothalamus (PVN) from male rats exposed to CIE vapor or air-exposed controls. Ca2+ signals were evaluated in response to norepinephrine (NE) with or without selective α-adrenergic receptor (αAR) or GluN2B-containing N-methyl-D-aspartate receptor (NMDAR) antagonists, followed by dexamethasone (DEX) to mock a pharmacological stress response. Expectedly, CIE rats had altered anxiety-like, rearing, grooming, and drinking behaviors. Importantly, NE-mediated reductions in Ca2+ event frequency were blunted in both CIE neurons and astrocytes. Administration of the selective α1AR antagonist, prazosin, reversed this CIE-induced dysfunction in both cell types. Additionally, the pharmacological stress protocol reversed the altered basal Ca2+ signaling profile of CIE astrocytes. Signaling changes in astrocytes in response to NE were correlated with anxiety-like behaviors, such as the grooming:rearing ratio, suggesting tripartite synaptic function plays a role in switching between exploratory and stress-coping behavior. These data show how CIE exposure causes persistent changes to PVN neuro-glial function and provides the groundwork for how these physiological changes manifest in behavioral selection.

Background: Hip resurfacing arthroplasty (HRA) and total hip arthroplasty (THA) are two treatment options for end-stage degenerative hip conditions. The objective of this single-center retrospective cohort study was to compare implant survival and patient-reported outcomes (PROs) in young patients (≤35 years) who underwent HRA or THA. Methods: All patients aged 35 years or younger who underwent HRA or THA with a single high-volume arthroplasty surgeon between 2004 and 2015 were reviewed. The sample included 33 THAs (26 patients) and 76 HRAs (65 patients). Five-year implant survival and minimum 2-year PROs were compared between patient cohorts. Results: Three patients in the THA group (9%) were revised within 5 years for instability (n = 1), squeaking (n = 1), or squeaking with a ceramic liner fracture (n = 1). No patients who underwent HRA were revised. The University of California, Los Angeles, activity score, modified Harris Hip score, and Hip Dysfunction and Osteoarthritis Outcome Scores for Joint Replacement increased by 74%, 64%, and 49%, respectively, among all patients. Compared to the HRA cohort, patients who underwent THA had lower preoperative and postoperative University of California, Los Angeles, activity, modified Harris Hip score, and Hip Dysfunction and Osteoarthritis Outcome Scores for Joint Replacement scores, yet there were no differences in the absolute improvements in any of the three measures between the two groups. Conclusions: Excellent functional outcomes were seen in young patients undergoing either HRA or THA. Although young patients undergoing THA started at lower preoperative baseline and postoperative PROs than patients undergoing HRA, both groups improved by an equal amount after surgery, suggesting that both HRA and THA afford a similar degree of potential improvement in a young population.

Background: Recent studies have examined the impact of the COVID-19 pandemic on the practice of total joint arthroplasty. A scoping review of the literature with compiled recommendations is a useful tool for arthroplasty surgeons as they resume their orthopedic practices during the pandemic. Methods: In June 2020, PubMed, Embase (Ovid), Cochrane Library (Wiley), Scopus, LitCovid, CINAHL, medRxiv, and bioRxiv were queried for articles using controlled vocabulary and keywords pertaining to COVID-19 and total joint arthroplasty. Studies were characterized by their region of origin, design, and Center of Evidence Based Medicine level of evidence. The identified relevant studies were grouped into 6 categories: changes to future clinical workflow, education, impact on patients, impact on surgeons, technology, and surgical volume. Results: The COVID-19 pandemic has had a significant impact on arthroplasty practice, including the disruption of the clinical teaching environment, personal and financial consequences for patients and physicians, and the drastic reduction in surgical volume. New pathways for clinical workflow have emerged, along with novel technologies with applications for both patients and trainees. Conclusions: The COVID-19 pandemic emphasizes the recent trend in arthroplasty toward risk stratification and outpatient surgery, which may result in improved clinical outcomes and significant cost-savings. Furthermore, virtual technologies are a promising area of future focus that may ultimately improve upon previous existing inefficiencies in the education and clinical environments.

SUMMARY: Effective implementation of noxious odor mitigation strategies requires a thorough understanding of production causes, a clear definition of harmful odor limits, and practical and effective methods to measure emissions into the environment. With the increasing demand for poultry products and consequently, intensification of production systems, there is a need for versatile odor remediation approaches. Hydrogen sulfide (H2S) is a pungent noxious gas associated with poultry farms. The presence and intensity of H2S from animal excreta is the result of the dynamic balance of its formation and reduction, under primarily anaerobic microbial fermentation conditions. Unlike other odors such as ammonia, H2S production and emission is a less researched subject. In the present study, a practical monitoring method was devised to evaluate the efficacy of a Bacillus-based microbial product as a strategy to remediate H2S emissions from poultry excreta. The evaluated excreta were preconditioned with distilled water and then incubated for 24 h under controlled temperature before odor measurement. Plastic containers were used as incubation units, and H2S diffusion tubes were used as the monitoring tool. Hydrogen sulfide emitted from the excreta of broilers fed the microbial product was reduced on average by 42.9% compared to a nonmicrobial fed group. Under the current trial conditions, H2S emitted from growing broiler chicken excreta was easily detectable and variation was low. Moreover, the method allowed dynamic and quick H2S monitoring that will further offer tools to the commercial poultry industry for evaluating the effectiveness of feeding strategies in odor remediation.

PI3K activity determines positive and negative selection of B cells, a key process for immune tolerance and B cell maturation. Activation of PI3K is balanced by phosphatase and tensin homolog (Pten), the PI3K’s main antagonistic phosphatase. Yet, the extent of feedback regulation between PI3K activity and Pten expression during B cell development is unclear. Here, we show that PI3K control of this process is achieved post-transcriptionally by an axis composed of a transcription factor (c-Myc), a microRNA (miR17-92), and Pten. Enhancing activation of this axis through overexpression of miR17-92 reconstitutes the impaired PI3K activity for positive selection in CD19-deficient B cells and restores most of the B cell developmental impairments that are evident in CD19-deficient mice. Using a genetic approach of deletion and complementation, we show that the c-Myc/miR17-92/Pten axis critically controls PI3K activity and the sensitivity of immature B cells to negative selection imposed by activation-induced cell death.

Summary: Inflammatory bowel disease (IBD) is prevalent, but the mechanisms underlying disease development remain elusive. We identify a role for the E3 ubiquitin ligase RNF5 in IBD. Intestinal epithelial cells (IECs) express a high level of RNF5, while the colon of Rnf5−/− mice exhibits activated dendritic cells and intrinsic inflammation. Rnf5−/− mice exhibit severe acute colitis following dextran sodium sulfate (DSS) treatment. S100A8 is identified as an RNF5 substrate, resulting in S100A8 ubiquitination and proteasomal-dependent degradation that is attenuated upon inflammatory stimuli. Loss of RNF5 from IECs leads to enhanced S100A8 secretion, which induces mucosal CD4+ T cells, resulting in Th1 pro-inflammatory responses. Administration of S100A8-neutralizing antibodies to DSS-treated Rnf5−/− mice attenuates acute colitis development and increases survival. An inverse correlation between RNF5 and S100A8 protein expression in IECs of IBD patients coincides with disease severity. Collectively, RNF5-mediated regulation of S100A8 stability in IECs is required for the maintenance of intestinal homeostasis. : Fujita et al. show that RNF5 regulation of S100A8 stability in intestinal epithelial cells defines the degree of pro-inflammatory response, culminating in severe intestinal inflammation following DSS treatment to Rnf5−/− mice. Neutralizing S100A8 antibodies attenuates acute colitis phenotypes, and inverse RNF5/S100A8 expression coincides with clinical severity in IBD patients. Keywords: ubiquitin ligase, RNF5, S100A8, intestinal inflammation, IBD, acute colitis, intestinal epithelial cells

Induced pluripotent stem cells (iPSCs) provide an inexhaustible source of cells for modeling disease and testing drugs. Here we develop a bioinformatic approach to detect differences between the genomic programs of iPSCs derived from diseased versus normal human cohorts as they emerge during in vitro directed differentiation. Using iPSCs generated from a cohort carrying mutations (PiZZ) in the gene responsible for alpha-1 antitrypsin (AAT) deficiency, we find that the global transcriptomes of PiZZ iPSCs diverge from normal controls upon differentiation to hepatic cells. Expression of 135 genes distinguishes PiZZ iPSC-hepatic cells, providing potential clues to liver disease pathogenesis. The disease-specific cells display intracellular accumulation of mutant AAT protein, resulting in increased autophagic flux. Furthermore, we detect beneficial responses to the drug carbamazepine, which further augments autophagic flux, but adverse responses to known hepatotoxic drugs. Our findings support the utility of iPSCs as tools for drug development or prediction of toxicity.

This study investigated the impact of glyphosate on bacterial communities and their degradation mechanisms in large-leaf tea soil, through exposure microcosm and enrichment culture experiments. Soils from three tea gardens in Yunnan, China, were used: two glyphosate-free (JM and KL) for microcosm study and one long-term exposed (G2) for enrichment culture experiment. The results revealed a two-phase degradation process with half-lives of 12.7 to 268 days, while the metabolite AMPA was notably persistent. The acidic conditions and high organic content of tea soils may retard glyphosate microbial availability and degradation. Glyphosate initially stimulated bacterial growth but led to abundance declines with prolonged exposure. It tended to enhance bacterial diversity at lower doses. Network complexity increased in JM soil where strong adsorption moderated glyphosate exposure, yet decreased in KL soil where weak adsorption enabled greater microbial-glyphosate interactions. Community structure analysis revealed soil-specific responses, with decreased Proteobacteria in JM soil and Actinobacteria in KL soil, while several phyla including Proteobacteria, Acidobacteriota, Chloroflexi, Myxococcota, and Verrucomicrobiota showed increased abundance. PICRUSt2 analysis indicated enhanced biosynthesis and cell growth pathways, while carbohydrate metabolism, nitrogen metabolism, and xenobiotics biodegradation pathways were reduced. LEfSe analysis identified potential degrading biomarkers primarily from Proteobacteria, Acidobacteriota, Myxococcota, Chloroflexi, and Actinobacteriota, suggesting their putative role in degradation. The enriched consortium G2 efficiently degraded 400 mg/L glyphosate within 7 days, with notable increases in Afipia, Dokdonella, and Cohnella abundance. This study provides insights into bacterial interactions with glyphosate in tea soils, suggesting strategies for contamination mitigation and environmental restoration., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

An imbalance in the microbiota-gut-brain axis exerts an essential effect on the pathophysiology of depressive and anxiety disorders. Our previous research revealed that the timing of inulin administration altered its effects on chronic unpredictable mild stress (CUMS)-induced anxiety and depression. However, it is still unclear if the gut-brain axis is primarily responsible for these effects. In this study, fecal microbiota transplantation (FMT) confirmed that inulin administration at different times alleviated CUMS-induced anxiety- and depression-like behaviors via the gut-brain axis. The time of administration seemed to modify the anxiolytic and antidepressant effects of inulin, and inulin intervention in the evening was more pronounced in inhibiting the inflammatory responses than that of morning inulin intervention. Serum metabolomics analysis showed that the main differential metabolites, including fenofibric acid, 4'-Hydroxyfenoprofen glucuronide and 5-(4-Hydroxybenzyl)thiazolidine-2,4-dione may be vital for the anxiolytic and antidepressant effects of different inulin treatment times. Our results suggested that inulin administration in the evening was more effective in alleviating the inflammatory responses and improving amino acids metabolism. This study provides a new potential link between the microbiota-gut-brain axis and chrono-nutrition, demonstrating that a more appropriate administration time results in a better intervention effect., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024. Published by Elsevier B.V.)

Chronic wound healing is often hindered by long-term inflammation and redox imbalance. Herbal medicine, with its rich medicinal components such as polysaccharides, flavonoids, phenolic acids, and small-molecule nutrients, has gained attention for its anti-inflammatory and antioxidant properties. Xanthium strumarium (XS) is a potent anti-inflammatory herb that has shown promise in treating conditions like rhinitis and may have specific benefits for chronic skin wounds. However, traditional XS preparations taken orally can have harmful effects on the liver and kidneys, limiting its clinical use. To date, there has been no documented method for the localized and sustained delivery of XS. This study introduces a timely development of a multifunctional hybrid hydrogel, incorporating bioactive XS extract and Gelatin Methacryloyl (GelMA), for the purpose of diabetic wound healing. In vitro experiments showed that XS/GelMA had strong anti-inflammatory and antioxidant effects through the Akt/mTOR pathway. The XS extract itself also significantly enhanced cell migration and angiogenesis. In vivo studies confirmed the superior wound healing properties of XS/GelMA. These findings suggest that XS-derived hydrogels have great potential for skin regeneration and tissue engineering applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

The C-3 and C-5 substituted 20-deoxyingenol monoesters are important active components in Euphorbiaceae plants. Nonetheless, their similar physical properties make them difficult to distinguish. The present study developed fast and efficient rules for identifying the esterification sites of 20-deoxyingenol based on a series of chemical syntheses of monoesters and literature research, utilizing NMR spectroscopy, optical rotation analysis, and chromatographic retention behavior. In addition, a series of 20-deoxyingenol ether derivatives, including 1,3,4-oxadiazole derivatives, were synthesized. The cytotoxic activities of 20-deoxyingenol derivatives were evaluated on A549 and HepG2 cell lines. Notably, 20-deoxyingenol 1,3,4-oxadiazole derivative 22 (IC 50  = 8.8 μM) exhibited significant anticancer activity against HepG2 cells with low toxicity to normal cells (IC 50  > 50 μM), making it a promising compound. We investigated the potential anticancer mechanism of compound 22 by examining protein expression changes in HepG2 cells using quantitative proteomics. Our findings indicated that 22 induced G1/S phase cell cycle arrest and, In a dose-dependent manner, inhibited CDK4 and CyclinD1 expression while upregulating P21. Moreover, 22 promoted the accumulation of autophagosomes and the proteins LC3 and PINK1, enhancing autophagy and mitophagy in HepG2 cells. Collectively, compound 22 might serve as a novel autophagy agonist with anticancer properties., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025. Published by Elsevier Inc.)

Objective: Electronic health record (EHR) systems contain a wealth of clinical data stored as both codified data and free-text narrative notes (NLP). The complexity of EHR presents challenges in feature representation, information extraction, and uncertainty quantification. To address these challenges, we proposed an efficient Aggregated naRrative Codified Health (ARCH) records analysis to generate a large-scale knowledge graph (KG) for a comprehensive set of EHR codified and narrative features., Methods: Using data from 12.5 million Veterans Affairs patients, ARCH first derives embedding vectors and generates similarities along with associated p-values to measure the strength of relatedness between clinical features with statistical certainty quantification. Next, ARCH performs a sparse embedding regression to remove indirect linkage between features to build a sparse KG. Finally, ARCH was validated on various clinical tasks, including detecting known relationships between entity pairs, predicting drug side effects, disease phenotyping, as well as sub-typing Alzheimer's disease patients., Results: ARCH produces high-quality clinical embeddings and KG for over 60,000 codified and narrative EHR concepts. The KG and embeddings are visualized in the R-shiny powered web-API 3 . ARCH achieved high accuracy in detecting EHR concept relationships, with AUCs of 0.926 (codified) and 0.861 (NLP) for similar EHR concepts, and 0.810 (codified) and 0.843 (NLP) for related pairs. It detected drug side effects with a 0.723 AUC, which improved to 0.826 after fine-tuning. Using both codified and NLP features, the detection power increased significantly. Compared to other methods, ARCH has superior accuracy and enhances weakly supervised phenotyping algorithms' performance. Notably, it successfully categorized Alzheimer's patients into two subgroups with varying mortality rates., Conclusion: The proposed ARCH algorithm generates large-scale high-quality semantic representations and knowledge graph for both codified and NLP EHR features, useful for a wide range of predictive modeling tasks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

The development of cellulose fabrics with good flame retardancy and durability has been a primary concern for in firefighting clothing. A recyclable ternary deep eutectic solvent (TDES) was used to prepare surface ammonium phosphate-modified cellulose fabrics (SACF). The incorporation of ammonium phosphate groups notably enhanced the durable flame retardancy of cellulose fabrics. The limiting oxygen index (LOI) of SACF could reach 48 %, and remain at 33.5 % even after 50 laundering cycles (LCs). And the TDES can be recovered and reused for 5 cycles and the SACF still remains flame retardancy. It also exhibited excellent antibacterial properties (with an antibacterial rate of 99.79 % against E. coli) and dyeability with cationic dyes. Moreover, the flame-retardant of the SACF did not decrease after dyed and showed both condensed-phase and gas-phase flame-retardant mechanisms during combustion. This method for producing flame-retardant cellulose fabrics with recyclable TDES effectively reduces the use of chemical reagents and holds promise for application in eco-friendly manufacturing and firefighting garments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025. Published by Elsevier B.V.)

The shortage of corneal donors and the limitations in tissue engineering grafts, such as biocompatibility and mechanical properties, pose significant challenges in corneal transplantation. Here, for the first time, we investigate the effect of Rho kinase inhibitor Y-27632 and a dual media culture approach, including proliferative media (M1) and stabilizing media (M2), on rabbit limbal epithelial stem cells (LESCs), aiming to explore the feasibility of constructing corneal cell sheets in vitro through auto-bioprinting and assessing their corneal wound healing capacity in vivo. Y-27632 has primarily demonstrated significantly enhanced LESCs growth, proliferation, and reduced apoptosis. The dual media culture approach combined with Y-27632 improved LESCs proliferation while maintaining stemness. In spheroid culture, Y-27632 decreased cell death and promoted proliferation. Immunofluorescent staining and RNA sequencing revealed upregulation of genes related to tight junctions and cell adhesion and downregulation of genes associated with aging and cell cycle. Using a bioprinter, LESC spheroids were auto-bioprinted onto a custom-made curved collagen membrane, creating a bioactive, transplantable, tissue-engineered anterior corneal sheet. Anterior superficial corneal transplantation with these LESC sheets significantly accelerated epithelial wound healing in rabbit limbal stem cell deficiency (LSCD) models. Overall, the integration of Y-27632, dual-media culture, and spheroid cell culture led to the development of a highly bioactive and therapeutically promising bio-ink derived from LESCs. Auto-bioprinting these LESC spheroids produced a bioactive, transplantable corneal cell sheet, presenting a promising therapeutic option for LSCD. STATEMENT OF SIGNIFICANCE: The renewal and wound healing of the corneal epithelium are essential for maintaining normal vision and refractive function. Limbal stem cell deficiency (LSCD) is a major cause of blinding keratopathy, and current treatment options are limited. In this study, for the first time, we developed a highly bioactive and therapeutically potent bio-ink for ocular surface regeneration by integrating Y-27632, a dual-media culture approach, and spheroid cell culture. Additionally, using auto-bioprinting technology, the limbal epithelial stem cell (LESC) spheroid bio-ink was precisely auto-bioprinted onto the curved surface of the corneal membrane, significantly accelerating corneal epithelial healing in an LSCD rabbit model., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025. Published by Elsevier Inc.)

Cancer is one of the most fatal diseases threatening public health globally, and tumor metastasis causes greater than 90 % of cancer-associated deaths, presenting huge challenges for detection and efficient treatment of various human cancers. Cancer stem cells (CSCs) are a rare population of cancer cells and increasing evidences indicated CSCs are the driving force of tumor metastasis. In this study, a p-AuNSs-assisted single-cell Raman spectra has been established, to extract and amplify of CSCs fingerprints with single cell sensitivity. The gathered rich molecular information was further assigned according to the characteristic Raman peaks, and the results revealed a huge difference in the expression of molecules between CSCs and cancer cells, such as nucleic acids, proteins, saccharides and lipids. Furthermore, multiple data analysis algorithms including PCA, SVM, RF and KNN, were employed to reveal the fundamental characteristics and classification of CSCs and cancer cells based on the whole p-AuNSs-assisted single-cell Raman spectra. This work is beneficial for not only providing deep insights for molecular behaviors of tumor metastasis based on CSCs, but also could obtain significant information to aid medical diagnosis and to design effective CSCs-based therapeutic intervention clinically., Competing Interests: Declaration of competing interest There are no conflicts to declare., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Ferroptosis, which depends on iron ions to generate reactive oxygen species (ROS), has been proved to be an effective strategy for cancer therapy. However, cells will initiate different programs, including reducing iron uptake and storing excess iron in ferritin, to lower the intracellular iron concentration. In this work, we reported a simple, one-pot method to synthesize bovine serum albumin stabilized MnFe 2 O 4 nanoparticles (MnFe 2 O 4 @BSA NPs) for ferroptosis therapy of cancer. Artemisinin (ART) and salinomycin (Sali), which could induce the degradation of ferritin and enhance the uptake by increasing binding protein IRP2 and transferrin receptor, were loaded onto the MnFe 2 O 4 @BSA NPs to strengthen the killing effect. The prepared MnFe 2 O 4 @BSA-ART/Sali (MnFe 2 O 4 /ART/Sali) NPs could significantly increase the cellular iron concertation, enhancing the ROS generation in cells. After intravenous injection, the MnFe 2 O 4 /ART/Sali NPs showed superior anti-tumor effects, with a tumor inhibition rate of 67.65 %. Hence, the hybrid nanocomposite indicated the combined effect of MnFe 2 O 4 , ART, and Sali, providing a platform to enhance ferroptosis therapy of cancer., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Flurochloridone (FLC) is a pyrrolidone herbicide used to control broad-leaved weeds in various crop fields. However, there is still a lack of comprehensive research on the environmental fate of the Qinghai-Tibet Plateau (QTP) and the toxicity of its potential transformation products (TPs). In this study, laboratory experiments were conducted to investigate the degradation kinetics, pathways, and toxicity of FLC's TPs. Nine TPs were identified in soil using Ultra-Performance Liquid Chromatography Quadrupole-Orbitrap Mass Spectrometry (UPLC-Q-Exactive Orbitrap MS) and Compound Discoverer software, employing suspect and nontarget screening strategies. The initial report of two TPs, TP204, and TP191 was confirmed through the acquisition or synthesis of their standards. High Performance Liquid Chromatography coupled with Tandem Mass Spectrometry (HPLC-MS/MS) was subsequently used for further quantification of these TPs in all samples under examination. The primary transformation reactions of FLC in the environment include oxidative dechlorination, reductive dechlorination, reductive defluorination, acetylation, and hydrolysis. Predictive assessments via ECOSAR, alongside empirical laboratory experiments, revealed that most novel TPs exhibit significantly lower acute toxicity towards Danio rerio, Daphnia magna Straus, and Algae compared to FLC. However, TP204 demonstrated neutral chronic toxicity towards Daphnia magna Straus and Green algae, potentially posing a latent threat to aquatic ecosystems. These results are crucial for elucidating the environmental fate of FLC, assessing environmental risks, and guiding scientific and reasonable use. This research holds significant importance for the ecological environment protection in the Tibetan Plateau region., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Several interventions have been investigated addressing perinatal depression; however, interventions have not been compared using both direct and indirect evidence. This study compared the relative effectiveness of all interventions that prevent depression (objective 1) or treat depression symptoms (objective 2) in perinatal women., Methods: Eight databases searched from their inception to March 2024. Inclusion criteria were randomised controlled trials in perinatal women participants evaluating any interventions that addressed prevention or treatment of depression. Two random-effects Bayesian network meta-analyses were conducted using studies with perinatal women participants reporting frequency of depression or reporting depression severity. For objective 1 odds ratios (OR) and objective 2 standardised mean differences (SMD) were used., Results: A total of 177 articles were included (n = 76 objective 1, n = 101 objective 2). For objective 1; mindfulness was the highest-ranking intervention (SCURA=88), with significant OR compared to treatment as usual (TAU) (OR=0.21; 0.06, 0.71 95 %CrI). Other interventions with significant OR, compared to TAU included: pharmacological treatments, CBT, physical activities, education, and collaborative care. For objective 2; interpersonal therapy plus massage was the highest-ranking intervention (SCURA=89) with significant SMD compared to TAU (SMD=-1.38; -2.54, -0.21). Other interventions with significant SMD compared to TAU: IPT, alternative therapies, physical activities, mindfulness, CBT, collaborative care, education, and enhanced TAU. Pharmacological treatments were effective compared to sham/placebo., Conclusion: CBT, mindfulness, physical activity, collaborative care, education, and pharmacological interventions were effective in addressing prevention and treatment of perinatal depression symptoms compared to control arms. Healthcare providers may consider offering these interventions to perinatal women., Competing Interests: Declaration of Competing Interest None., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Salmonella infection is an acute and systemic disease of poultry, primarily affecting young birds. The mortality rate of chicken within one week of age can reach up to 40 %. Surviving individuals may become carriers of the bacteria, leading to latent infections that can result in bacterial residues in meat and egg products, posing serious threats to human food safety and health. Antibiotic therapy is one of the most conventional treatments for Salmonella infections in birds. However, the current abuse of antibiotics has accelerated the mutation of pathogenic bacteria to generate antibiotic-resistant strains. Thus, the effectiveness of treatment with antibiotics alone is gradually diminishing. To address this threat, researchers have explored the use of natural products to enhance the immune system of poultry for preventing Salmonella infections. This study aims to provide a comprehensive review, systematically summarizing recent research progress of the application of natural medicines on poultry Salmonella infection., Competing Interests: Declaration of competing interest The authors declare that they do not have any competing interests. There were no financial and personal relationships with other people or organizations that could have inappropriately influenced the study. There were no professional or other personal interests of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled., (Copyright © 2024. Published by Elsevier Inc.)

Hemp seed cake (HSC) (Cannabis sativa L.) is a high-quality plant-derived protein source rich in polyunsaturated fatty acid (PUFA). To assess the effects of HSC addition in diets on the growth and meat quality in broiler chicken, a total of 240 female three-yellow chickens (50 days of age) were randomly assigned to four groups and fed with varying levels of HSC (0% (HSC0), 5% (HSC5), 10% (HSC10), and 20% (HSC20)) for 9 weeks. As a result, the daily feed intake, weight gain and feed conversion efficiency were significantly increased in the HSC20 group. Moreover, the meat quality traits, including the meat colour, water-holding capacity, intramuscular fat content, and proportion of n-3 PUFA significantly improved, and the expression of lipid synthesis genes, were increased in the HSC20 group. Meanwhile, the development of immune organs and the anti-inflammatory capabilities were enhanced in the HSC20 group. In addition, the blood lipid of chicken was reduced by improving the lipid metabolism in the HSC20 group. Therefore, adding 20% HSC in the feed had a notable effect on the growth, antioxidant and immune capabilities, blood lipid metabolism, and meat performance of the female three-yellow chickens. These findings provide significant information for improving the production performance of broiler chickens through the effective utilization of HSC., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

L-asparaginase (L-ASNase) can hydrolyze L-asparagine, a precursor to acrylamide, thereby reducing toxic acrylamide formation in fried foods. Currently, commercial L-ASNases are primarily produced by wild-type (WT) filamentous fungi; however, these enzymes often exhibit rapid activity loss during high-temperature processing due to limited thermal stability. In this study, we screened a thermostable L-ASNase gene from thermophile bacteria and expressed it in Aspergillus niger to reduce acrylamide content in French fries. Initially, four genes encoding thermostable L-ASNases were selected and integrated into the A. niger genome via non-homologous end joining. Among these, the L-ASNase gene tzi from Thermococcus zilligii was successfully expressed in A. niger, yielding an extracellular activity of 114 U·mg -1 . The recombinant enzyme (An-Tzi) displayed the same optimal temperature and pH as its WT counterpart but exhibited superior catalytic efficiency, likely due to the efficient post-translational modifications in A. niger. To further enhance expression, the tzi gene was integrated into the amylase (amyA) locus of the A. niger genome using the CRISPR-Cas9 system, resulting in increased activity of 128 U·mg -1 . Additionally, various lengths of the highly expressed glucoamylase (glaA) protein from A. niger AG11 were fused to the N-terminus of the Tzi. Notably, fusing the 500-amino-acid catalytic domain of glaA led to a substantial 3.3-fold increase in enzyme activity. Despite the metabolic stress induced by high-level expression of glaA, supplementing the culture medium with metal ions and sophorose resulted in an extracellular activity of 486.74 U·mg -1 , the highest reported yield of L-ASNase in shake flasks. Finally, applying the An-Tzi to French fries achieved a 32 % greater reduction in acrylamide compared to the commercial enzyme. Overall, the recombinant A. niger strain expressing thermostable An-Tzi demonstrates significant potential for industrial applications targeting acrylamide reduction in fried and baked foods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

We evaluated the prognostic and therapeutic significance of measurable residual disease (MRD) during remission induction in pediatric acute lymphoblastic leukemia (ALL) patients. In the CCCG-ALL-2015 protocol, 7640 patients were categorized into low-, intermediate-, or high-risk groups based on clinical and genetic features. Final risk classification was determined by MRD assessed via flow cytometry on Days 19 and 46 of remission induction, with additional intensified chemotherapy for Day 19 MRD ≥1%. B-ALL patients with negative MRD (<0.01%) on Day 19 or Day 46 had significantly better 5-year event-free survival (EFS) than those with MRD 0.01-0.99%, who in turn had better EFS than patients with MRD ≥1%. Provisional low-risk patients with Day 19 MRD ≥1% but negative Day 46 MRD, reclassified as intermediate-risk, had comparable 5-year EFS to low-risk patients with Day 19 MRD 0.3-0.99% and negative Day 46 MRD (82.5% vs. 83.0%) and better EFS than provisional low-risk patients with MRD on both days (83.0% vs. 72.6%, P<0.001). Similarly, provisional intermediate-risk B-ALL patients with Day 19 MRD ≥1% but negative Day 46 MRD, who received additional therapy, had better 5-year EFS compared to those with Day 19 MRD between 0.3-0.99% (70.7% vs. 53.0%, P<0.001). Among low-risk patients with negative Day 46 MRD, those with negative Day 19 MRD had superior EFS compared to those with positive Day 19 MRD (91.7% vs. 86.1%, P<0.001). Optimal use of Day 19 MRD could improve individualized treatment and outcomes. Chinese Clinical Trial Registry (ChiCTR-IPR-14005706)., (Copyright © 2024 American Society of Hematology.)

Cervical cancer is a significant public health issue for women, with human papillomavirus (HPV) infection rates exhibiting regional variations throughout China. This study examined data from a cohort of 24,588 rural women who engaged in cervical cancer screening in Luonan County from 2021 to 2023, utilizing high-risk HPV (hrHPV) testing methodologies. The findings revealed an overall HPV positivity rate of 14.18 %, with hrHPV positivity rates of 13.99 % in 2021, 12.97 % in 2022, and 15.32 % in 2023. Infection rates increased with age, showing significant differences among age groups. Types 52, 16, and 58 were the most prevalent hrHPV types. Single-type infections were more common than multiple infections, with prevalent combinations such as HPV16 + HPV52, HPV52 + HPV58, and HPV51 + HPV16. The results highlight the imperative for improved screening, especially for women aged 60 to 64 residing in rural areas, and emphasize the importance of HPV vaccination and targeted screening for the prevention and eventual eradication of cervical cancer., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Author(s).)

Avian pathogenic Escherichia coli (APEC) is a bacterial pathogen that threatens poultry reproduction by inciting systemic inflammation and leading to chicken colibacillosis. The endocannabinoid system (ECS) is an immunomodulator system that regulates inflammatory responses. In this study, we aimed to investigate the anti-inflammatory effect of paeoniflorin on APEC-infected HD11 cells and its underlying mechanism. The results showed that paeoniflorin significantly reduced the expression levels of pro-inflammatory factors (IL-1β, IL-6, TNF-α), M1-type macrophage-associated markers (IL-12, iNOS, CD86), and chemokines (CXCL8, CXCL12, CCL1, CCL5, CCL17). Additionally, paeoniflorin significantly reduced the expression of MAGL and restored that of DAGL and CB 2 R, thereby activating the ECS. Furthermore, we found that paeoniflorin and CB 2 R exhibited stable conformations through molecular docking and molecular dynamics simulations. The addition of the CB 2 R inhibitor AM630 notably diminished paeoniflorin's inhibitory effects on the phosphorylation levels of proteins in the NF-κB signaling pathway and on inflammatory responses. These results indicate that the anti-inflammatory effects of paeoniflorin on APEC-induced HD11 cells may be mediated by the inhibition of the NF-κB signaling pathway through the activation of CB 2 R. This work has the potential to provide a new agent for the control and prevention of chicken colibacillosis, as well as contribute to modern research in traditional Chinese medicine., (Copyright © 2024. Published by Elsevier Inc.)

Dust storms have a profound impact on air quality, atmospheric chemistry, and human well-being by carrying vast amounts of particles over distances of thousands of kilometers. However, the overall characteristics of these dust events and their influence on secondary pollution in the northern China region are not yet well understood, due to a lack of long-term, comprehensive observations and objective identification techniques. Based on principal component analysis combined with high-time-resolution observations of particulate matter components, here we developed a robust method to identify dust storm events and identified 14 dust events in Beijing in 2019. We further classified these 14 events into two distinct types using Lagrangian particle dispersion models and backward trajectory analysis. The first type (Type I, 9 cases) is characterized by synoptic patterns in Mongolia, originating from the north and directly impacting the Beijing area. The second type (Type II, 5 cases) involves air masses from the north or northwest that temporarily pass through polluted regions south of Beijing before being carried back into the city. Consistently, during Type I dust events, we observed a sharp decrease in secondary inorganic aerosols (SIA) from 65 % to 7 %, as well as in the sulfur oxidation ratio (SOR) and nitrogen oxidation ratio (NOR) (from 0.52 to 0.19, and 0.27 to 0.018 respectively). In contrast, during Type II dust events, SIA concentrations increased by 91 %, along with an increase in SOR (1.7 %), NOR (69 %), and f 44 /f 43 (3.0 %), suggesting an enhancement of secondary aerosol formation resulting from the interaction between dust aerosols and gaseous anthropogenic pollutants. Our results demonstrate that dust events and the sub-type of dust events can be identified in an objective manner using the protocol developed in this study and the dynamics should be considered when discussing impacts of dust events on atmospheric chemistry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Uveal melanoma (UM) poses a significant lethality, with approximately 50% of those developing metastases surviving less than one year. In the progression of UM, vasculogenic mimicry (VM) induced by hypoxia plays a pivotal role, which also partially explains the resistance of UM to anti-angiogenic therapies. Nevertheless, the crucial molecular mechanisms underlying VM in the progression of UM remain unclear. We identified ubiquitin conjugating enzyme E2 G2 (UBE2G2) as a critical suppressor through transcriptomic sequencing and metastasis correlation screening. In UM, hypoxia-induced VM and metastasis are markedly exacerbated by UBE2G2 knockdown and significantly alleviated by its overexpression. Mechanistically, UBE2G2 directly binds to galectin 3 binding protein (LGALS3BP) and forms a complex with the E3 ubiquitin ligase tripartite motif containing 38 (TRIM38), facilitating ubiquitination-mediated degradation of LGALS3BP at the K104 residue. Furthermore, UBE2G2 inhibits oncogenic phenotypes by inactivating intracellular PI3K/AKT signaling and reprogramming the tumor microenvironment. Therefore, targeting intercellular and intracellular molecular mechanisms of the hypoxia-UBE2G2-LGALS3BP axis may contribute to developing various therapeutic strategies for UM., Competing Interests: The authors declare no potential conflicts of interest., (© 2024 The Authors.)

Background: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are severe adverse drug reactions with extensive keratinocyte death. Carbamazepine (CBZ), the most commonly implicated drug in SJS/TEN, is metabolized by the cytochrome P450 enzyme 3A4 (CYP3A4) into carbamazepine-10,11-epoxide (CBZE) in the liver. While CD8 + cytotoxic T cells play an important role in SJS/TEN, the underlying mechanism of exuberant immune response by CD8 + T cells in these conditions remains incompletely understood., Objectives: To examine the expression of NLRP3 inflammasome and their skin migration in CBZE-induced SJS/TEN., Methods: The expression of the NLRP3 inflammasome complex in skin lesions, sera, and blister fluids of SJS/TEN patients were analyzed by immunohistochemistry and enzyme-linked immunosorbent assay. NLRP3 formation and CD8 + T cell activation status and their functions were examined by immunoblotting, immunofluorescence, and chemotaxis assays., Results: The expression of the NLRP3 inflammasome complex was greatly increased in skin lesions of SJS/TEN patients. Moreover, IL-1β and IL-18 levels in sera and blister fluids of SJS/TEN patients were approximately 3-fold higher than those in healthy individuals, with a linear correlation between IL-1β levels and disease activity. CBZE induced NLRP3 inflammasome formation, upregulated CXCL9/CXCL10 levels, and activated CD8 + cytotoxic T cell functions via IL-1β/IL-1R or IL-18/IL-18R signaling in SJS/TEN keratinocytes, which promoted CD8 + cytotoxic T cell migration in SJS/TEN patients., Conclusion: This study showed that CBZE promoted NLRP3 inflammasome formation and strengthened the activation and function of CD8 + cytotoxic T cells in the skin, which contributed to the initiation and progression of SJS/TEN., (Copyright © 2024 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.)

The fungal pathogen Setosphaeria turcica (S. turcica) causes northern corn leaf blight (NCLB), resulting in significant yield and economic losses in maize. To elucidate the metabolic pathways essential for its pathogenicity, we investigated the metabolome of S. turcica during appressorium development, a critical stage for host infection. Our analysis indicated a substantial enrichment of sphingosine and related compounds during this phase. The application of chemical inhibitors to disrupt sphingolipid metabolism confirmed their pivotal role in appressorium formation and pathogenicity. Additionally, silencing of the serine palmitoyl transferase (Spt) core subunit gene StLCB2 led to significant alterations in fungal morphology and growth, accompanied by changes in cell membrane integrity, surface hydrophobicity, melanin, and sphingosine synthesis. These findings underscore the importance of sphingolipids in the pathogenicity of S. turcica and suggest that targeting specific components of the sphingolipid pathway could aid in developing novel fungicides or genetically engineered maize varieties with increased resistance to NCLB., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

HClO/ClO - and H 2 S are two kinds of momentous biological small molecules in the organism, and the redox balance between them is considerable for the physiological and pathological properties of organisms. Hence, it is very crucial to monitor the redox course between HClO and H 2 S. Herein, a reversible fluorescent sensor (IPSe) for ClO - and H 2 S was firstly constructed with the selenium atom as the response site and the dicyanoisophorone as the fluorophore. The sensor IPSe could detect ClO - with good selectivity and sensitivity due to the oxidation reaction of the selenium atom triggered by ClO - . The recognition of IPSe to hypochlorite induced a hypsochromic shift of the absorption maximum from 420 nm to 380 nm. IPSe exhibited the prominent low detection limit of 55.3 nM for detecting ClO - , accompanied by distinct fluorescent attenuation. Moreover, H 2 S could efficiently return the fluorescence of the IPSe solution to the original level by H 2 S reducing selenoxide. The experimental results show that the suggested method has high precision and accuracy for the detection of ClO - and H 2 S. The applications in real water samples, beverages and cell imaging verified that the IPSe was capable of monitoring the changes in the concentration of ClO - /H 2 S, which indicates that it is of great meaning to survey the biological functions of ClO - and H 2 S via IPSe., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Elevated ammonium (NH 4 -N) contents in groundwater are a global concern, yet the mobilization and enrichment mechanisms controlling NH 4 -N within riverside aquifers (RAS) remain poorly understood. RAS are important zones for nitrogen cycling and play a vital role in regulating groundwater NH 4 -N contents. This study conducted an integrated assessment of a hydrochemistry dataset using a combination of hydrochemical analyses and multivariate geostatistical methods to identify hydrochemical compositions and NH 4 -N distribution in the riverside aquifer within Central Yangtze River Basin, ultimately elucidating potential NH 4 -N sources and factors controlling NH 4 -N enrichment in groundwater ammonium hotspots. Compared to rivers, these hotspots exhibited extremely high levels of NH 4 -N (5.26 mg/L on average), which were mainly geogenic in origin. The results indicated that N-containing organic matter (OM) mineralization, strong reducing condition in groundwater and release of exchangeable NH 4 -N in sediment are main factors controlling these high concentrations of NH 4 -N. The Eh representing redox state was the dominant variable affecting NH 4 -N contents (50.17 % feature importance), with Fe 2+ and dissolved organic carbon (DOC) representing OM mineralization as secondary but important variables (26 % and 5.11 % feature importance, respectively). This study proposes a possible causative mechanism for the formation of these groundwater ammonium hotspots in RAS. Larger NH 4 -N sources through OM mineralization and greater NH 4 -N storage under strong reducing condition collectively drive NH 4 -N enrichment in the riverside aquifer. The evolution of depositional environment driven by palaeoclimate and the unique local environment within the RAS likely play vital roles in this process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: The utility of repeat ablation for premature ventricular contractions (PVCs) after prior ablation failure is not clear., Objective: The purpose of this study was to assess the outcomes of repeat ablation and the use of different techniques in patients who failed prior PVC ablation., Methods: We reviewed 239 consecutive patients who underwent PVC ablation. When standard endocardial ablation with normal or half-normal saline failed, we considered an advanced ablation technique. Acute success was defined as abolition of the target PVC. Clinical and procedural findings, PVC origins, and acute and follow-up outcomes were compared in those with and without a prior failed ablation procedure., Results: Of 239 patients, 75 (31%) had failed a prior ablation procedure, and they more often had left ventricular outflow tract PVCs. Despite failing prior ablation, repeat standard ablation was acutely successful in 59% of patients, and 75% of these patients had long-term success. Acute standard ablation success rate was lower and long-term recurrence rate was higher than in patients without prior ablation (59% vs 95%, P <.001; and 29% vs 17%, P <.05, respectively). Of the 31 repeat standard procedures that again failed, advanced techniques were performed in 23 (16 needle, 5 epicardial, 2 simultaneous ablation) and were acutely successful in 16 (70%) with long-term success in 14 (45%). Overall long-term success for patients with prior failed standard ablation was 71%., Conclusion: Although success is lower for patients with prior failed ablation, repeat ablation seems reasonable for many, and the use of advanced techniques increased success to 71% in this group., Competing Interests: Disclosures Dr Tokutake was supported by a scholarship from the Japanese Heart Rhythm society. Dr Uetake has received support from Mitsukoshi Health and Welfare Foundation and Fukuda Foundation for Medical Technology. Dr Hasegawa has received support from the International Rotary Fellowship of Healthcare Professionals. Dr Richardson is a consultant for Medtronic, Johnson & Johnson, and Philips; and has received research funding from Medtronic and Abbott. Dr Kanagasundram has received speaking honoraria from Johnson & Johnson. Dr Michaud has received honoraria from Boston Scientific, Abbott, Johnson & Johnson, and Medtronic; and is a consultant for Boston Scientific, Abbott, Johnson & Johnson, and GE. Dr Stevenson is a co-holder of a patent for irrigated needle ablation with rights assigned to MGH Brigham Hospital; and has received speaking honoraria from Abbott, Boston Scientific, Biotronik, Johnson & Johnson. All other authors have no conflicts to disclose., (Copyright © 2024 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.)

As nature-based solutions, urban park plant communities play a pivotal role in regulating urban carbon cycles, alleviating global climate change, and fostering sustainable urban development. However, the factors influencing the carbon sink efficiency of plant communities in urban parks within temperate monsoon climate zones have not been fully investigated. This study used multivariate heterogeneous data to evaluate plant communities' carbon storage (CS) and annual carbon sequestration (ACS) in 25 urban parks across different biotope types in Jinan, a city located in China's temperate monsoon climate zone. The driving mechanisms affecting carbon sink efficacy were revealed using Spearman correlation, regression, principal component analyses, and structural equation modeling. Results demonstrated that: 1) Closed broadleaf multi-layer green space has significant carbon sink potential compared to other vegetation structures. 2) The carbon sink efficiency of the plant communities negatively correlated with the sky view factor and planting layout density. Three-dimensional green quantity (3DGQ), the ratio of trees and shrubs, species richness, and vertical structures positively correlated with plant communities' carbon storage and sequestration. 3) Whether increasing 3DGQ, the ratio of trees and shrubs, or the total number of individuals of all species, there is a certain threshold bottleneck in enhancing the carbon sink benefits of plant communities. 4) Plant community structure, species composition, and species diversity influenced carbon sink efficiency, collectively forming the first principal component. The 3DGQ affected carbon sink efficiency as the second principal component. Synergistic effects existed among these driving factors, jointly explained 64.3 % and 90.1 % of the CS and ACS of plant communities, respectively. Optimization design strategies for different plant communities in urban parks were proposed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Competing Interests: Disclosures The authors have no conflicts of interest to disclose.

Wide used guided bone regeneration (GBR) membrane materials, such as collagen, Teflon, and other synthesized polymers, present a great challenge in term of integrating the mechanical property and degradation rate when addressing critical bone defects. Therefore, inspired by the distinctive architecture of fish scales, this study utilized epigallocatechin gallate to modify decellularized fish scales following biomimetic mineralization to fabricate a GBR membrane that mimics the structure of lamellar bone. The structure, physical and chemical properties, and biological functions of the novel GBR membrane were evaluated. Results indicate that the decellularized fish scale with 5 remineralization cycles (5R-E-DCFS) exhibited a composite and structure similar to natural bone and had a special functionally gradient mineral contents character, demonstrating excellent mechanical properties, hydrophilicity, and degradation properties. In vitro, the 5R-E-DCFS membrane exhibited excellent cytocompatibility promoting Sprague-Dawley (SD) rat bone marrow mesenchymal stem cell proliferation and differentiation up-regulating the expression of osteogenic-related genes and proteins. Furthermore, in vivo, the 5R-E-DCFS membrane promoted the critical skull bone defects of SD rats repairment and regeneration. Therefore, this innovative biomimetic membrane holds substantial clinical potential as an ideal GBR membrane with mechanical properties for space-making and suitable degradation rate for bone regeneration to manage bone defects., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Rhizoctonia solani is a soil-borne pathogen with 14 anastomosis groups (AGs), and different subgroups are genetically diverse. However, the genetic factors contributing to the pathogenicity of the fungus have not been well characterized. In this study, the genome of R. solani AG1-ZJ was sequenced. As the result, a 41.57 Mb draft genome containing 12,197 putative coding genes was obtained. Comparative genomic analysis of 11 different AGs revealed conservation and unique characteristics between the AGs. Furthermore, a novel effector family containing a 68 amino acid conserved domain unique in basidiomycetous fungi was characterized. Two effectors containing the conserved domain in AG4-JY were identified, and named as RsUEB1 and RsUEB2. Furthermore, the spray-induced gene silencing strategy was used to generate a dsRNA capable of silencing the conserved domain sequence of RsUEB1 and RsUEB2. This dsRNA can significantly reduce the expression of RsUEB1 and RsUEB2 and the pathogenicity of AG4-JY on foxtail millet, maize, rice and wheat. In conclusion, this study provides significant insights into the pathogenicity mechanisms of R. solani. The identification of the conserved domain and the successful use of dsRNA silencing of the gene containing the conserved domain will offer a new strategy for controlling sheath blight in cereal crops., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)