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Avian reovirus (ARV) is the causative agent of avian viral arthritis and causes significant economic losses to the global poultry industry. For clinical diagnosis, detecting ARV-specific antibodies is crucial. We successfully expressed the ARV-σC protein in insect cells using the baculovirus expression vector system, achieving an expression level of approximately 200 mg/L. We developed an indirect enzyme-linked immunosorbent assay (iELISA) using the ARV-σC protein as a coating antigen to detect antibodies against it. The inter-batch and intrabatch coefficients of iELISA variation were less than 10%. Its sensitivity (1:12,800 diluted in serum) was 4 times higher than that of the indirect immunofluorescence assay (IFA; 1:3200 diluted in serum), and it showed no cross-reactivity with antibodies against other common avian viruses (such as Infectious bursal disease virus, Newcastle disease virus). The practicality of the iELISA was further evaluated using clinical samples. 300 clinical sera from chickens vaccinated with the ARV attenuated vaccine and 20 SPF sera were tested using both the iELISA and the IFA, demonstrating a 100% conformity rate. In conclusion, these results suggest that the iELISA developed in this study is a rapid, sensitive, and specific method that could serve as an effective diagnostic tool for monitoring and controlling avian viral arthritis., Competing Interests: DISCLOSURES All authors disclosed no relevant relationships., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Flame-retardant epoxy resins with tough, transparent, ultraviolet shielding, and low dielectric properties have fascinating prospects in electronic and electrical applications, but it is still challenging at present. In this work, a bio-based macromolecule was synthesized from vanillin (a lignin derivative), phenyl dichlorophosphate, 9,10-dihydro-9-oxa-10-phosphaphenanthrene 10-oxide (DOPO), and poly(propylene glycol) bis(2-aminopropyl ether). The bio-based macromolecule, namely, MFR, was designed and added to the epoxy resin (EP). The cured EP containing 15 wt% MFR (i.e., EP/MFR15) exhibits excellent flame retardancy with an Underwriter Laboratory 94 (UL-94) V-0 rating and a limiting oxygen index (LOI) of 29.2 %. Furthermore, the peak heat release rate (PHRR) and total heat release rate (THR) are drastically reduced by 59.5 % and 40.7 %, respectively. Meanwhile, EP/MFR15 shows 20.3 % and 43.8 % improvements in tensile strength and toughness, respectively. Moreover, MFR simultaneously endows EP with accessional ultraviolet shielding performance and low dielectric constant without sacrificing transparency. This work provides a promising strategy for fabricating a bio-based macromolecular flame retardant and preparing a high-performance EP composite with versatile 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 © 2024. Published by Elsevier B.V.)

This study investigated the mechanisms of microbial growth and metabolism during biofilm cultivation in the biofilm sequencing batch reactor (BSBR) process for phosphate (P) enrichment. The results showed that the sludge discharge was key to biofilm growth, as it terminated the competition for carbon (C) source between the nascent biofilm and the activated sludge. For the tested reactor, after the sludge discharge on 18 d, P metabolism and C source utilization improved significantly, and the biofilm grew rapidly. The P concentration of the recovery liquid reached up to 157.08 mg/L, which was sufficient for further P recovery via mineralization. Meta-omics methods were used to analyze metabolic pathways and functional genes in microbial growth during biofilm cultivation. It appeared that the sludge discharge activated the key genes of P metabolism and inhibited the key genes of C metabolism, which strengthened the polyphosphate-accumulating metabolism (PAM) as a result. The sludge discharge not only changed the types of polyphosphate-accumulating organisms (PAOs) but also promoted the growth of dominant PAOs. Before the sludge discharge, the necessary metabolic abilities that were spread among different microorganisms gradually concentrated into a small number of PAOs, and after the sludge discharge, they further concentrated into Candidatus_Contendobacter (P3) and Candidatus_Accumulibacter (P17). The messenger molecule C-di-GMP, produced mostly by P3 and P17, facilitated P enrichment by regulating cellular P and C metabolism. The glycogen-accumulating organism (GAO) Candidatus_Competibacter secreted N-Acyl homoserine lactones (AHLs), which stimulated the secretion of protein in extracellular polymeric substances (EPS), thus promoting the adhesion of microorganisms to biofilm and improving P metabolism via EPS-based P adsorption. Under the combined action of the dominant GAOs and PAOs, AHLs and C-di-GMP mediated QS to promote biofilm development and P enrichment. The research provides theoretical support for the cultivation of biofilm and its wider application., 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.)

Objective: Laparoscopic surgical skill assessment and machine learning are often inaccessible to low-and-middle-income countries (LMIC). Our team developed a low-cost laparoscopic training system to teach and assess psychomotor skills required in laparoscopic salpingostomy in LMICs. We performed video review using AI to assess global surgical techniques. The objective of this study was to assess the validity of artificial intelligence (AI) generated scoring measures of laparoscopic simulation videos by comparing the accuracy of AI results to human-generated scores., Design: Seventy-four surgical simulation videos were collected and graded by human participants using a modified OSATS (Objective Structured Assessment of Technical Skills). The videos were then analyzed via AI using 3 different time and distance-based calculations of the laparoscopic instruments including path length, dimensionless jerk, and standard deviation of tool position. Predicted scores were generated using 5-fold cross validation and K-Nearest-Neighbors to train classifiers., Setting: Surgical novices and experts from a variety of hospitals in Ethiopia, Cameroon, Kenya, and the United States contributed 74 laparoscopic salpingostomy simulation videos., Results: Complete accuracy of AI compared to human assessment ranged from 65-77%. There were no statistical differences in rank mean scores for 3 domains, Flow of Operation, Respect for Tissue, and Economy of Motion, while there were significant differences in ratings for Instrument Handling, Overall Performance, and the total summed score of all 5 domains (Summed). Estimated effect sizes were all less than 0.11, indicating very small practical effect. Estimated intraclass correlation coefficient (ICC) of Summed was 0.72 indicating moderate correlation between AI and Human scores., Conclusions: Video review using AI technology of global characteristics was similar to that of human review in our laparoscopic training system. Machine learning may help fill an educational gap in LMICs where direct apprenticeship may not be feasible., (Copyright © 2023 Association of Program Directors in Surgery. Published by Elsevier Inc. All rights reserved.)

Microbial transglutaminase (mTG) has become a powerful tool for manufacturing antibody-drug conjugates (ADCs). It enables site-specific conjugation by catalyzing formation of stable isopeptide bond between glutamine (Q) side chain and primary amine. However, the downstream impact of mTG-mediated conjugation on ADC product quality, especially on high molecular weight (HMW) size variant formation has not been studied in a systematic manner. This study investigates the mechanisms underlying the formation of HMW size variants in mTG-mediated ADCs using size exclusion chromatography (SEC) and liquid chromatography-mass spectrometry (LC-MS). Our findings revealed that the mTG-mediated glutamine and lysine (K) crosslinking is the primary source of the increased level of HMW size variants in the ADCs. In the study, two monoclonal antibodies (mAbs) with glutamine engineered for site-specific conjugation were used as model systems. Based on the LC-MS analysis, a single lysine (K56) in the heavy chain (HC) was identified as the major Q-K crosslinking site in one of the two mAbs. The HC C-terminal K was observed to crosslink to the target Q in both mAbs. Quantitative correlation was established between the percentage of HMW size variants determined by SEC and the percentage of crosslinked peptides quantified by MS peptide mapping. Importantly, it was demonstrated that the level of HMW size variants in the second ADC was substantially reduced by the complete removal of HC C-terminal K before conjugation. The current work demonstrates that crosslinking and other side reactions during mTG-mediated conjugation needs to be carefully monitored and controlled to ensure process consistency and high product quality of the final ADC drug product., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Yimeng Zhao, Sunnie Kim, Xiang Zheng, Se Hyun Kim, Amy Han, Tse-Hong Chen, Serena Wang, Jieqiang Zhong, Haibo Qiu, Ning Li reports financial support was provided by Regeneron Pharmaceuticals Inc. Yimeng Zhao, Sunnie Kim, Xiang Zheng, Haibo Qiu has patent pending to Yimeng Zhao, Sunnie Kim, Xiang Zheng, Haibo Qiu., (Copyright © 2023 American Pharmacists Association. Published by Elsevier Inc. All rights reserved.)

The disposal of hazardous municipal solid waste incineration (MSWI) fly ash is a challenge nowadays. Recently, the re-utilization of MSWI fly ash by converting it to useful zeolite-containing materials has attracted attention. However, the zeolitic products fabricated from MSWI fly ash are usually of low quality and rarely reported to be applied for photocatalysis. In this study, valuable zeolites (e.g., NaP1) are synthesized from MSWI fly ash via a modified microwave-assisted hydrothermal method. The key parameters for the hydrothermal method including temperature, duration, the amount of additive, and water volume, are investigated and optimized. Specifically, increasing the hydrothermal temperature can promote the synthesis of zeolitic materials; a relatively long hydrothermal duration is essential to accomplish the assembly of zeolites; the addition of Na 2 SiO 3 can increase the precursor for the fabrication of zeolites; the water volume makes little influence on the crystal style of products. Eventually, the hydrothermal condition of 180 °C, 1 h, 0.5 g Na 2 SiO 3 , and 10 mL water is suggested based on the energy consumption and the quality of zeolites. The product containing zeolite NaP1 from such a condition is further applied to degrade methylene blue by photocatalysis. The removal rate has reached 96% within 12 h, which dramatically surpasses that of the raw fly ash (38%). Such excellent photocatalytic performance is attributed to the 10-fold increased surface area (24.864 m 2  g -1 ) and active metal elements embedding in the zeolite structures., 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 © 2022 Elsevier Inc. All rights reserved.)

The Si and Al in municipal solid waste incineration fly ash (MSWI FA) can be utilized for zeolite fabrication, which can improve the application value of the products. This study focuses on the fabrication of zeolite from MSWI FA by microwave-assisted hydrothermal (MH) treatment. The effects of magnetic stirring time, Na 2 SiO 3 dosage, MH time, and NaOH solution concentration on the crystallization of zeolite NaP1 from MSWI FA are systematically analyzed. The synthetic products are analyzed through spectroscopic and mineralogical methods. The results show that zeolite NaP1 with high crystallinity (51.68 %) can be fabricated by magnetic stirring and MH treatment, and the cation exchange capacity (CEC) of the product can reach a value of 2.58 meq/g, which is approximately 133 times that of the CEC of MSWI FA. The Si/Al ratio plays a decisive role in the zeolite NaP1 synthesis, and a Na 2 SiO 3 dosage of 30 wt% is adopted for zeolite NaP1 fabrication. A NaOH concentration of 1 M is sufficient for zeolite NaP1 synthesis. Additionally, the zeolite NaP1 content is found to obviously increase with increasing MH time from 0.5 h to 2 h. To demonstrate the feasibility of the method provided in this study, the optimal experimental condition is employed for various MSWI FAs, and zeolite NaP1 and analcime are fabricated successfully. The leachability of heavy metals for the synthetic products was evaluated, which met the requirements for pollution control. The BET surface area and total pore volume of zeolite NaP1 fabricated at optimal condition are 61.42 m 2 /g and 0.44 cm 3 /g, respectively. The adsorption capacity of zeolite NaP1 for Cu 2+ ion and methylene blue are determined to be 84.65 mg/g and 84.55 mg/g, respectively, indicating zeolite NaP1 is a potential adsorbent for cation ion and dyes. This study provides an environmentally friendly scheme for the utilization of MSWI FA., 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 © 2022 Elsevier B.V. All rights reserved.)

Municipal solid waste incineration fly ash (MSWI FA) is categorized as a hazardous waste, which demands environmentally acceptable treatment due to its easy leachability toxic of heavy metals. This study investigated an innovative and improved method for stabilization/solidification (S/S) of MSWI FA with coal gangue based geopolymer by the addition of active calcium content. The specimen with addition of calcium oxide up to 10 % reached the compressive strength of 2.14 MPa at 28 d. The addition of 30 % calcium oxide resulted in the highest immobilization efficiencies of Cd (98.96 %) and Pb (99.19 %). X-ray Diffraction (XRD), Fourier Transform Infrared Spectrometry (FTIR), Scanning Electron Microscope (SEM), and thermogravimetric (TG) analysis indicated the generation of calcium-containing hydration products was promoted after the improvement of calcium content in binder. Heavy metals were stabilized through the chemical adsorption and ions exchange of amorphous hydration products. On the whole, this study illustrated that the incorporation of active calcium content can improve efficiently S/S of hazardous ash waste such as MSWI FA., 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 © 2022 Elsevier B.V. All rights reserved.)

Fly ash is an incineration byproduct of thermal power plants. Due to the complex composition of fly ash, improper disposal will seriously harm the ecological environment. Therefore, how to effectively use fly ash to safely and environmentally replace landfills is a worldwide concern. Considering the high silicon and aluminum contents in fly ash, it has the potential to synthesize zeolite, which has a wide range of applications in sewage treatment, gas adsorption, etc. Therefore, the synthesis of zeolites from fly ash is consistent with the theme of sustainable development. The synthesis mechanism of zeolite, various synthetic methods of zeolite from fly ash and their advantages and disadvantages was introduced in detail. In addition, combined with the current research hotspots, the application of synthetic zeolite from fly ash in the fields of sewage treatment and gas adsorption was introduced. Finally, the future development prospects and research directions of synthetic zeolite from fly ash to improve the utilization rate of fly ash were considered., 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 © 2022 Elsevier B.V. All rights reserved.)

Bipolar disorder (BD) is associated with increased suicidal behavior. Understanding the neural features of suicide attempts (SA) in patients with BD is critical to preventing suicidal behavior. The prefrontal cortex (PFC) is a key region related to SA. In this study, forty BD patients with a history of SA (BD+SA), 70 BD patients without a history of SA (BD-SA), and 110 individuals in a healthy control (HC) group underwent structural magnetic resonance imaging (MRI) and resting-state functional MRI. We used voxel-based morphometry (VBM) and amplitude of low frequency fluctuations (ALFF) techniques to examine the gray matter volumes (GMVs) and ALFF values in the PFC. Compared with the HC group, both the BD+SA and BD-SA groups had lower GMVs and higher ALFF values in the medial PFC (MPFC), ventral PFC (VPFC), and dorsolateral PFC (DLPFC). The ALFF values in the MPFC, VPFC, and DLPFC in the BD+SA group were significantly higher than those in the BD-SA group. These findings suggest that BD patients with SA have intrinsic activity abnormalities in PFC regions. This provides potentially identifiable neuroimaging markers in BD patients with SA that could be used to increase our understanding of suicidal behavior., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Capillary zone electrophoresis-electrospray ionization-mass spectrometry (CZE-ESI-MS) was used for analysis of reduced antibodies. We first developed a simple protocol to condition commercial linear-polyacrylamide coated capillaries for use in top-down proteomics. We then suspended reduced antibodies in a solution of 35% acetic acid, 50% acetonitrile in water. Heavy and light chains were baseline resolved within 10 min and with 3-30 µg/mL detection limits using a 0.1% aqueous formic acid background electrolyte. Quintuplicate runs of a two-antibody mixture produced relative standard deviations of ∼1% in migration time and 10% in peak amplitudes. Resolution was further improved for the two-antibody mixture by using 5% acetic acid as the background electrolyte, highlighting the potential of capillary electrophoresis-mass spectrometry for analysis of antibody mixtures., (Copyright © 2015 Elsevier B.V. All rights reserved.)

We coupled capillary zone electrophoresis (CZE) with an ultrasensitive electrokinetically pumped nanospray ionization source for tandem mass spectrometry (MS/MS) analysis of complex proteomes. We first used the system for the parallel reaction monitoring (PRM) analysis of angiotensin II spiked in 0.45mg/mL of bovine serum albumin (BSA) digest. A calibration curve was generated between the loading amount of angiotensin II and intensity of angiotensin II fragment ions. CZE-PRM generated a linear calibration curve across over 4.5 orders of magnitude dynamic range corresponding to angiotensin II loading amount from 2amole to 150fmole. The relative standard deviations (RSDs) of migration time were <4% and the RSDs of fragment ion intensity were ∼20% or less except 150fmole angiotensin II loading amount data (∼36% RSD). We further applied the system for the first bottom up proteomic analysis of a human cell line using CZE-MS/MS. We generated 283 protein identifications from a 1h long, single-shot CZE MS/MS analysis of the MCF7 breast cancer cell line digest, corresponding to ∼80ng loading amount. The MCF7 digest was fractionated using a C18 solid phase extraction column; single-shot analysis of a single fraction resulted in 468 protein identifications, which is by far the largest number of protein identifications reported for a mammalian proteomic sample using CZE., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Myotonic dystrophy type 1 (DM1) is a multi-systemic disease with no established treatment to date. Small, cell-permeable molecules hold the potential to treat DM1. In this study, we investigated the association between protein kinase C (PKC) signaling and splicing of sarcoplasmic reticulum Ca(2+)-ATPase1 (SERCA1). Our aim was to clarify the mechanisms underlying the regulation of alternative splicing, in order to explore new therapeutic strategies for DM1. By assessing the splicing pattern of the endogenous SERCA1 gene in HEK293 cells, we found that treatment with phorbol 12-myristate 13-acetate (PMA) regulated SERCA1 splicing. Interestingly, treatment with PMA for 48 h normalized SERCA1 splicing, while treatment for 1.5h promoted aberrant splicing. These two responses showed dose dependency and were completely abolished by the PKC inhibitor Ro 31-8220. Furthermore, repression of PKCβII and PKCθ by RNAi mimicked prolonged PMA treatment. These results indicate that PKC signaling is involved in the splicing of SERCA1 and provide new evidence for a link between alternative splicing and PKC signaling., (Copyright © 2012 Elsevier Inc. All rights reserved.)