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Lina Gao,1 Lei Zhang,1 Fengjun He,1 Jing Chen,1 Meng Zhao,1 Simin Li,1 Hao Wu,1 Yumeng Liu,1 Yinan Zhang,1,2 Qineng Ping,3 Lihong Hu,1,2 Hongzhi Qiao1,2,4 1Jiangsu Key Laboratory for Functional Substance of Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, People’s Republic of China; 2State Key Laboratory Cultivation Base for TCM Quality and Efficacy, School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023, People’s Republic of China; 3Department of Pharmaceutics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, 210009, People’s Republic of China; 4Jiangsu Engineering Research Center for Efficient Delivery System of TCM, Nanjing University of Chinese Medicine, Nanjing, 210023, People’s Republic of ChinaCorrespondence: Hongzhi Qiao; Lihong HuState Key Laboratory Cultivation Base for TCM Quality and Efficacy, School of Pharmacy, Nanjing University of Chinese Medicine, 138 Xianlin Avenue, Nanjing, 210023, People’s Republic of ChinaEmail qiaohz@njucm.edu.cn; lhhu@njucm.edu.cnBackground: BF211, a derivative of bufalin (BF), shows significantly improved solubility and potent antitumor efficiency compared to BF. Unfortunately, the unwanted toxicity such as cardiotoxicity caused by unspecific distribution has hindered its clinical use.Methods: PEGylated BF211 liposomes (BF211@Lipo) were designed and optimizely prepared based on the pre-prescription research. In vitro and in vivo cardiotoxicity was evaluated. In vivo pharmacokinetics and biodistribution of BF211@Lipo were investigated. In vivo antitumor activity and toxicity were evaluated in HepG2 cell xenograft models. The rapid-release triggered by Poloxamer 188 (P188) was assessed in vitro and in vivo.Results: The optimized BF211@Lipo displayed a spherical morphology with a size of (164.6 ± 10.3) nm and a high encapsulation efficiency of (93.24 ± 2.15) %. The in vivo concentration–time curves of BF211 loaded in liposomes showed a prolonged half-life in plasma and increased tumor accumulation. No obvious abnormality in electrocardiograms was observed in guinea pigs even at 9 mg/kg. Moreover, to improve the efficient release of BF211@Lipo, a surfactant-assisted rapid-release strategy was developed, and the release-promoting mechanism was revealed by the fluorescence resonance energy transfer (FRET) and fluorescence nanoparticle tracking analysis (fl-NTA) technology. Sequential injection of BF211@Lipo and P188 could ignite the “cold” liposomes locally in tumor regions, facilitating the burst release of BF211 and enhancing the therapeutic index.Conclusion: Our progressive efforts that begin with preparation technology and dosage regimen enable BF211 to like a drug, providing a promising nano platform to deliver the cardiac glycosides and alleviate the side effects by decreasing unspecific biodistribution.Keywords: bufalin derivative, liposome, rapid-release, antitumor, cardiotoxicity

Ying Xu,1 Shengzhe Lu,1 Qi Liu,2 Yun Hong,3 Bohui Xu,4 Qineng Ping,5 Xuefeng Jin,5 Yan Shen,5 Thomas J Webster,6 Yuefeng Rao3,7 1Department of Pharmaceutics, School of Pharmacy, Jiangsu University, Zhenjiang, 212013, China; 2Department of Dermatology, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA; 3Department of Pharmacy, School of Medicine, The First Affiliated Hospital, Zhejiang University, Hangzhou 310003, China; 4Department of Pharmacy, School of Pharmacy, Nantong University, Nantong 226001, China; 5Department of Pharmaceutics, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China; 6Department of Chemical Engineering, Northeastern University, Boston, MA 02115, USA; 7Department of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China Background: In order to develop a promising carrier for the oral delivery of proteins and peptide drugs, a novel bioadhesive nanocarrier of chitosan (CTS) derivatives coated with poly (n-butyl) cyanoacrylate nanoparticles (PBCA-NPs) was prepared in this study.Methods: Three different thymopentin (TP5)-loaded nanoparticles were prepared in the present study. TP5-PBCA-NPs were developed by modifying an emulsion polymerization method, and CTS and chitosan–glutathione (CG) derivative-coated PBCA nanoparticles were obtained from the electrostatic interactions between CTS or CG with negatively charged PBCA nanoparticles.Results: The particle sizes of TP5-PBCA-NPs, TP5-CTS-PBCA-NPs, and TP5-CG-PBCA-NPs were 212.3±6.9, 274.6±8.2, and 310.4±7.5 nm, respectively, while the respective zeta potentials were –22.6±0.76, 23.3±1.2, and 34.6±1.6 mV with encapsulation efficiencies of 79.37%±2.15%, 74.21%±2.13%, and 72.65%±1.48%, respectively. An everted intestinal ring method indicated that drug stability was remarkably improved after incorporation into the nanoparticles, especially the CG-coated nanoparticles. The mucus layer retention rates for CTS- and CG-coated nanoparticles were 1.43 and 1.83 times that of the uncoated nanoparticles, respectively, using ex vivo mucosa. The in vivo mucoadhesion study illustrated that the transfer of uncoated PBCA-NPs from the stomach to the intestine was faster than that of CTS-PBCA-NPs and CG-PBCA-NPs, while the CG-PBCA-NPs presented the best intestinal retentive characteristic.Conclusion: In summary, this study demonstrated the feasibility and benefit of orally delivering peptide drugs using novel CTS derivative-coated nanoparticles with optimal stability and bioadhesive properties. Keywords: chitosan derivatives, PBCA nanoparticles, thymopentin, stability, bioadhesive

Juan Li,1 Shuangyan Tian,1 Qi Tao,2 Yawen Zhao,1 Ruyi Gui,1 Fan Yang,1 Lingquan Zang,3 Yanzhong Chen,4 Qineng Ping,5 Dongzhi Hou1 1Department of Pharmaceutics, Guangdong Provincial Engineering Center of Topical Precise Drug Delivery System, Guangdong Pharmaceutical University, Guangzhou, People’s Republic of China; 2CAS Key Laboratory of Mineralogy and Metallogeny & Guangdong Provincial Key Laboratory of Mineral Physics and Materials, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou, People’s Republic of China; 3Department of Pharmacology, College of Pharmacy, Guangdong Pharmaceutical University, Guangzhou, People’s Republic of China; 4Guangdong Provincial Key Laboratory of Advanced Drug Delivery Systems, Guangdong Pharmaceutical University, Guangzhou, People’s Republic of China; 5College of Pharmacy, China Pharmaceutical University, Nanjing, People’s Republic of China Background: To date, the rapid clearance from ocular surface has been a huge obstacle for using eye drops to treat glaucoma, since it has led to the short preocular residence time and low bioavailability. Methods: The novel nanoparticles (NPs) were designed for topical ophthalmic controlled drug delivery system through intercalating the BH into the interlayer gallery of Na-montmorillonite (Na+Mt) and then further enchasing chitosan nanoparticles. The resulting nanoparticles had a positive charge (+29±0.18 mV) with an average diameter of 460±0.6 nm. Results: In vitro study of drug release profiles suggested controlled release pattern. The irritation experiment analysis on both human immortalized cornea epithelial cell (iHCEC) and chorioallantoic membrane-trypan blue staining (CAM-TBS) showed good tolerance for ocular tissues. It was interestingly found that the nanoparticles could enter into iHCEC from the result of cellular uptake experiment measured by confocal layer scan microscopy (CLSM). Meanwhile, multilayered iHCEC was used to simulate the barrier of corneal epithelial cells for in vivo preocular retention capacity study, which suggested that BH-Mt/CS NPs could prolong the retention time in comparison with BH solution. The ocular pharmacokinetics studied by microdialysis sampling technique showed that AUC0-t and MRT0-t of BH-Mt/CS NPs were 1.99-fold and 1.75-fold higher than those of BH solution, indicating higher bioavailability. Moreover, the study of blood drug concentration, few researchers have reported, showed that low level drug could enter into blood, suggesting lower systematic side effect. Importantly, pharmacodynamics studies suggested that BH-Mt/CS NPs could make a significant decreased intraocular pressure on glaucomatous rabbits. Conclusion: Inspired by these advance of montmorillonite/chitosan nanoparticles, we envision that the BH-Mt/CS NPs will be a potential carrier for BH, opening up the possible applications in glaucoma therapy. Keywords: montmorillonite, glaucoma, chitosan, betaxolol hydrochloride, preocular retention

Yurui Xu,1 Sajid Asghar,2 Shiya Gao,1 Zhipeng Chen,3 Lin Huang,1 Lining Yin,1 Qineng Ping,1 Yanyu Xiao11Department of Pharmaceutics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, People’s Republic of China; 2Faculty of Pharmaceutical Sciences, Government College University Faisalabad, Faisalabad, Pakistan; 3Department of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, People’s Republic of China Abstract: The aim of this study was to evaluate the potential of polyelectrolyte complex nanoparticles (PENPs) based on hyaluronic acid/chitosan hydrochloride (HA/HCS) for co-loading mitoxantrone (MTO) and verapamil (VRP) to overcome multidrug resistance in breast tumors. PENPs co-loaded with MTO and VRP (MTO-VRP-PENPs) were affected by the method of preparation, molecular weight of HA, mass ratios and initial concentrations of HA/HCS, pH, and drug quantities. Optimized MTO-VRP-PENPs were ~209 nm in size with a zeta potential of approximately −24 mV. Encapsulation efficiencies (%) of MTO and VRP were 98.33%±0.27% and 44.21%±8.62%, respectively. MTO and VRP were successfully encapsulated in PENPs in a molecular or amorphous state. MTO-VRP-PENPs showed significant cytotoxicity in MCF-7/ADR cells in contrast to MTO-loaded PENPs (MTO-PENPs). The reversal index of MTO-VRP-PENPs was 13.25 and 10.33 times greater than that of the free MTO and MTO-PENPs, respectively. In conclusion, MTO-VRP-PENPs may serve as a promising carrier to overcome tumor drug resistance.Keywords: hyaluronic acid, chitosan hydrochloride, mitoxantrone hydrochloride, verapamil hydrochloride, co-delivery, multidrug resistance

Cihui Tian,1 Sajid Asghar,2 Yifan Wu,1 Zhipeng Chen,3 Xin Jin,1 Lining Yin,1 Lin Huang,1 Qineng Ping,1 Yanyu Xiao1 1Department of Pharmaceutics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, China; 2Faculty of Pharmaceutical Sciences, Government College University Faisalabad, Faisalabad, Pakistan; 3Department of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, China Abstract: The expression of multiple receptors on intestinal epithelial cells enables an actively targeted carrier to significantly enhance the oral delivery of payloads. Conjugating the receptors’ ligands on the surfaces of a particulate-delivery system allows site-specific targeting. Here, we used taurocholic acid (TCA) as a ligand for uptake of nanostructured lipid carriers (NLCs) mediated by a bile-acid transporter to improve oral bioavailability of curcumin (Cur). First, synthesis of TCA–polyethylene glycol 100–monostearate (S100-TCA) was carried out. Then, the physical and chemical properties of S100-TCA-modified Cur-loaded NLCs (Cur-TCA NLCs) with varying levels of S100-TCA modifications were investigated. Small particle size (90%), drug loading (about 3%), negative ζ-potential (–7 to –3 mV), and sustained release were obtained. In situ intestinal perfusion studies demonstrated improved absorption rate and permeability coefficient of Cur-TCA NLCs. Depending on the degree of modification, Cur-TCA NLCs displayed about a five- to 15-fold higher area under the curve in rats after oral administration than unmodified Cur NLCs, which established that the addition of S100-TCA to the NLCs boosted absorption of Cur. Further investigations of TCA NLCs might reveal a bright future for effective oral delivery of poorly bioavailable drugs. Keywords: curcumin, nanostructured lipid carriers, taurocholic acid, oral administration, pharmacokinetics, bioavailability

The different scientific studies have identified the beneficial physiological role and effects of functional foods (FFs). No doubt the knowledge of nutrition is primarily important in addressing human physical and physiological health along with human wellness. The study is planned to explore the consumers' knowledge about the different attributes of FFs and their consumption pattern. A purposive multistage random sampling design was used for the selection of 400 consumers of FFs from four major cities of Pakistan. The shopping centers and consumers were randomly selected. The analysis demonstrates that about more than 30 percent of the consumers had knowledge to a great extent of perceived quality, appearance package, organoleptic, functionality, price and brand attributes and about 40 percent of the consumers had knowledge to some extent of these attributes. Analysis reflects that from 33 to 86 percent of consumers were using often modified cereals, oats, bread, canola oil, orange juice, grape juice, modified yogurt, milk fortified with vitamins, green tea, energy drinks, ginger, and turmeric. It is suggested that creating awareness and building confidence about FFs among consumers will be useful for promotion of FFs in Pakistan. As the elder family members are more concerned about their health, they should motivate their children to adopt good food habits which are imperative to improve their quality of life. The FFs providers and manufactures should equip their R&D department with the latest development in functional foods sector through coordinating food scientists, university food experts and industrialists for manufacturing the specific food products which are beneficial for human health. [ABSTRACT FROM AUTHOR]

The application of urea fertilizer has always been associated with heavy N losses therefore, improving its efficiency is critical to minimize economic and environmental losses associated with its application. One possible approach to improve the nitrogen losses from the surface applied urea is to coat it with sulfur, urease inhibitor (agrotain) and other biodegradable materials. A pot experiment was carried out using two different textured: silt loam and clay loam soil to-assess the effect of sulfur and urease inhibitor (agrotain) on dry matter yield, N uptake and grain quality at the green house of the Zhejiang University, Hangzhou, China during 2011-12. The experiment consisted of treatments arranged in a completely randomized block design (CRD) with 3 replications. Urea coated with sulfur and urease inhibitor (agrotain) was applied at 60 kg N ha-1 in 2 splits. The results showed that sulfur and urease inhibitor (agrotain) coated urea significantly increased the dry matter yield, N uptake and grain quality of rice cultivars over granular urea (GU) and control treatment applied split, particularly in silty loam soil than clay loam soil. The N release was more pronounced in silty loam (SL) soil as compared to clay loam (CL) soil where, the effect of the inhibitors was less evident. Japonica cultivar (xinshui 123) showed its superiority over indica cultivar (9311) in all the studied parameters. It is concluded from these results that controlled released urea fertilizer improved dry matter yield by 55-68% ha-1, enhanced N uptake upto 39.4% and grain protein percentage from 5.8%-14.9% as compared to granular urea and control treatment. The improvement in rice dry matter yield and grain quality caused by sulfur and urease inhibitor (agrotain) coating may have been due to increased N use efficiency with split application in these two different textured soils. [ABSTRACT FROM AUTHOR]

To develop a long-acting injectable huperzine A-PLGA microsphere for the chronic therapy of Alzheimer's disease, the microsphere was prepared by using an o/w emulsion solvent extraction evaporation method based on a series of formulation design of the emulsion. The dialysis method was used for release analysis. The encapsulation efficiency and release amount of the microspheres were determined by a UV/VIS spectrophotometer. The morphology of the microspheres was observed by scanning electron microscopy. The distribution of the drug within microspheres was observed by a confocal laser scanning microscope. The results indicated that the PLGA 15?000 microspheres possessed a smooth and round appearance with average particle size of 50?µm or so. The encapsulation percentages of microspheres prepared from PLGA 15?000, 20?000 and 30?000 were 62.75%, 27.52% and 16.63%, respectively. The drug release percentage during the first day decreased from 22.52% of PLGA 30?000 microspheres to 3.97% of PLGA 15?000 microspheres, the complete release could be prolonged to 3 weeks. The initial burst release of microspheres with higher molecular weight PLGA could be explained by the inhomogeneous distribution of drug within microspheres. The encapsulation efficiency of the microspheres improved as the polymer concentration increased in the oil phase and PVA concentration decreased in the aqueous phase. The burst release could be controlled by reducing the polymer concentration. Evaporation temperature had a large effect on the drug release profiles. It had better be controlled under 30°C. Within a certain range of particle size, encapsulation efficiency decreased and drug release rate increased with the reducing of the particle size. [ABSTRACT FROM AUTHOR]

The purpose of this study was to investigate the solubilization capacity of sodium cholate/lecithin-mixed micelles and to evaluate the potential of mixed micelles as a carrier of cyclosporine A for intravenous infusion. The mixed micelles were prepared by coprecipitation technique. The formulation components and preparation procedures, which may affect the solubilization of cyclosporine A, were studied. The dilution stability of cyclosporine A-containing mixed micelles was investigated. Pharmacokinetic behaviors of mixed micelles in rabbits after intravenous infusion were compared with Sandimmun®. Results showed the strategies to increase the solubility of cyclosporine A include lowering the molar ratio of sodium cholate to lecithin, increasing the concentration of lecithin, and reducing the ionic strength of the dispersion medium and temperature. The largest solubility was found to be 5.42±0.16 mg/ml. The leakage of mixed micelles in 5%glucose (5.84%) was much less than that in saline solution (36.7%). The relative bioavailability of mixed micelles versus Sandimmun®was 112±20%, and statistical analysis demonstrated both preparations were bioequivalent. Sodium cholate/lecithin-mixed micelles are promising carriers in the intravenous delivery of cyclosporine A, considering their capability of large-scale production and low-toxic property. [ABSTRACT FROM AUTHOR]

Background: White granulated sugar, a significant commodity in bulk trade and widely used raw material, plays a crucial role in the food, energy, and medicine industries. A certified reference material (CRM) of white granulated sugar provides a valuable tool for monitoring and maintaining the safety and quality of white granulated sugar and other related products. The colorimetric value, conductivity ash, sucrose content, and reducing sugar content serve as essential factors that determine the quality of white granulated sugar., Objective: Develop a CRM of grade one white granulated sugar, conduct a comprehensive study to assess its homogeneity and stability, and determine the designated property values., Methods: Collaborative certification across 11 laboratories was applied to validate its property values and evaluate corresponding uncertainties., Results: The designated property values and expanded uncertainties (k = 2) determined to be 75.6 ± 2.8 IU, 0.0100 ± 0.0004 g/100g, 99.71 ± 0.08 g/100g, and 0.0226 ± 0.0024 g/100g for colorimetric value, conductivity ash, sucrose content, and reducing sugar content, respectively., Conclusion: The reference material (RM) was sufficiently homogeneous between and within bottles and remained stable for up to 20 months., Highlights: The developed RM of the grade one white granulated sugar could be an effective support for method validation, capability validation, quality control, and metrological traceability in the sugar and food industries., (© The Author(s) 2024. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Although biochar (BC)-assisted microbial electrolysis cells (MEC) has been shown to improve anaerobic digestion (AD) performance of waste activated sludge (WAS), the underlying mechanisms remain unclear. This study conducted an in-depth investigation into the mechanism based on analyses of extracellular polymeric substances (EPS) characteristics, enzyme activities and multi-omics. The results showed that compared with the control group, methane production improved by 16.73 %, 21.32 %, and 29.37 % in the BC, MEC, and BC-assisted MEC (BC-MEC) groups, respectively. The reconfiguration of the protein secondary structure increased the hydrophobicity of the EPS, thereby promoting microbial aggregation. In addition, partial least-squares path modeling (PLS-PM) and mantel test based on the enzyme activity and multi-omics analyses revealed that the promotional effect of MEC on the hydrolysis of WAS was superior to that of BC, while BC was more advantageous in promoting electron transfer and biofilm formation regulated by quorum sensing. The synergistic effects of BC and MEC were exemplified in the BC-MEC group. g_norank_Aminicenantales responsible for the hydrolysis of WAS was enriched (29.6 %), and the activities of hydrolytic enzymes including α-glucosidases and proteases were increased by 29.1 % and 43.6 %, respectively. Further, the expressions of genes related to acyl homoserine lactones (AHLs) and diffusible signal factor (DSF) in quorum sensing systems, as well as the genes related to hydrogenase involved in electron transfer (mbhJKL, hyfB-JR, hypA-F, and hoxFHUY), were up-regulated in the BC-MEC group. This facilitated electron transfer and microbial communication, consequently enhancing methane production. This research significantly advances the understanding of the mechanism by which BC-assisted MEC enhances AD performance and provides valuable insights into strategies for improving energy recovery from WAS., 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 Ltd. All rights reserved.)

Background: N6-methylandenosine-related long non-coding RNAs (m 6 A-related lncRNAs) play a crucial role in the cancer progression and immunotherapeutic efficacy. The potential function of m 6 A-related lncRNAs signature in cervical cancer has not been systematically clarified., Methods: RNA-seq and the clinical data of cervical cancer were extracted from The Cancer Genome Atlas. All of the patients were randomly classified into training and testing cohorts. The m 6 A-related lncRNAs prognostic model was constructed by LASSO regression using data in the training cohort.The predictive value of the signature was validated in the whole cohort and testing cohort. Cervical cancer patients were divided into low- and high-risk subgroups by the median value of risk scores. Kaplan-Meier analysis, principal-component analysis (PCA), functional enrichment annotation, and nomogram were used for further evaluation. We also examined the immune response and potential drug sensitivity targeting this model., Results: Seventy-nine prognostic m 6 A-related lncRNAs were screened. The risk model comprising four m 6 A-related lncRNAs (AL139035.1, AC015922.2, AC073529.1, AC008124.1) was identified and verified as an independent prognostic predictor of cervical cancer. A nomogram based on age, tumor grade, clinical stage, TNM stage, and four m 6 A-related lncRNAs risk signatures was generated. It displayed good accuracy and reliability in predicting the overall survival of patients with CC. Based on our risk model, cervical cancer patients with potential immunotherapy benefits from the candidate drugs could be effectively screened., Conclusion: The four m 6 A-related lncRNAs signature may provide new targets and allow the prediction of immunotherapy response, which can assist developing individualized treatment for cervical cancer., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

In this study, a laboratory-scale improved constructed rapid infiltration (imCRI) system with non-saturated and saturated layers was constructed, and corn cobs as solid carbon source were added to the saturated layer to enhance the removal of nitrogen. Combined analyses of metagenomics and metabolomics were conducted to elucidate the nitrogen removal mechanism in the imCRI system. The results showed that the hydraulic load significantly influenced the treatment performance of the imCRI system, and a hydraulic load of 1.25 m 3 /(m 2 ⋅d) was recommended. Under optimal conditions, the imCRI system using simulated wastewater achieved average removal efficiencies of 97.8 % for chemical oxygen demand, 85.7 % for total nitrogen (TN), and 97.6 % for ammonia nitrogen. Metagenomic and metabolomic analyses revealed that besides nitrification and denitrification, dissimilatory nitrate reduction to ammonium (DNRA), anammox, etc., are also involved in nitrogen metabolism in the imCRI system. Although nitrification was the predominant pathway in the non-saturated layer, aerobic denitrification also occurred, accounting for 22.59 % of the TN removal. In the saturated layer, nitrogen removal was attributed to synergistic effects of denitrification, DNRA and anammox. Moreover, correlation analysis among nitrogen removal, functional genes and metabolites suggested that metabolites related to the tricarboxylic acid cycle generated from the glycolysis of corn cobs provided sufficient energy for denitrification. Our results can offer a promising technology for decentralized wastewater treatment with stringent nitrogen removal requirements, and provide a foundation for understanding the underlying nitrogen transformation and removal mechanism., 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 Ltd. All rights reserved.)

In this study, the transformation mechanisms of extracellular polymeric substances (EPS) during ultraviolet/peracetic acid (UV/PAA) disinfection were elucidated based on multiple molecular-level analyses. After UV/PAA disinfection, the contents of soluble EPS (S-EPS), loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS) were reduced by 70.47 %, 57.05 % and 47.46 %, respectively. Fluorescence excitation-emission matrix-parallel factor and Fourier transform ion cyclotron resonance mass spectrometry analyses showed that during UV/PAA disinfection, EPS was transformed from the state characterized by high aromaticity, low saturation and low oxidation to the one with reduced aromaticity, increased saturation and higher oxidation. Specifically, sulfur-containing molecules (CHOS, CHONS, etc.) in EPS were converted into highly saturated and oxidized species (such as CHO), with the aromaticity index (AI mod ) decreasing by up to 53.84 %. Molecular characteristics analyses further indicated that saturation degree, oxidation state of carbon and molecular weight exhibited the most significant changes in S-EPS, LB-EPS and TB-EPS, respectively. Additionally, mechanistic analysis revealed that oxygen addition reaction was the predominant reaction for S-EPS (+O) and TB-EPS (+3O) (accounting for 31.78 % and 36.47 %, respectively), while the dealkylation was the main reaction for LB-EPS (29.73 %). The results were consistent with functional groups sequential responses analyzed by Fourier transform infrared and two-dimensional correlation spectroscopy, and were further verified by density functional theory calculations. Most reactions were thermodynamically feasible, with reaction sites predominantly located at functional groups such as CO, CO, CN and aromatic rings. Moreover, metabolomics analysis suggested that changes in metabolites in raw secondary effluent during UV/PAA disinfection were strongly correlated with EPS transformation. Our study not only provides a strong basis for understanding EPS transformation during UV/PAA disinfection at molecular-level but also offers valuable insights for the application this promising disinfection 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 Ltd. All rights reserved.)

Low enzyme activity is one of the disadvantages of immobilized laccase. In this study, waste Cu 2+ -loaded activated carbon (Cu-AC) was successfully used in preparing a novel composite support，cellulose / Cu 2+ -loaded activated carbon beads (C / Cu-AC)， and effectively boosted immobilized laccase activity. To achieve optimum conditions for immobilization of laccase, the immobilization time, pH and laccase concentration were examined. The highest immobilized laccase activity (34.21 U/g) was achieved under optimum conditions (T = 4 h, pH = 4, C = 5 g/L), which was increased by 35.86 % compared to control. In addition, the immobilized laccase showed an outstanding performance in thermostability and reusability compared to free laccase. Moreover, the degradation of BPA by immobilized laccase was carried out, and the optimum degradation conditions were explored. Under such conditions: concentration of BPA was 75 mg / L and pH = 4, t = 1 h, T = 50 °C，the removal yield of BPA reached a maximum of 79.88 %. Therefore, the utilization of waste Cu-CA is a powerful method to boost immobilized laccase activity and creating a new way to high value treatment of waste Cu-CA., 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.)