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This study introduces a novel method for detecting free glycidol and total free monochloropropanediol (MCPD) in fish and krill oil. Before analysis on high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS), p -(dimethylamino)phenol was used for derivatization of these compounds, enabling the sensitive determination of these contaminants. The sample preparation procedure includes a simple, efficient pretreatment using NaCl aqueous solution extraction and C18 sorbent cleanup (for demulsification), distinguishing glycidol from MCPD under varied reaction conditions for derivatization (weak acidic and strong alkaline aqueous environments). This approach shows broad linearity from 1 to at least 256 ng·mL -1 , improved sensitivity compared to standard GC-MS methods, with the limit of detection (LOD) and limit of quantification (LOQ) for MCPD and glycidol in both oil samples verified at 0.5 ng·mL -1 and 1 ng·mL -1 , respectively. Different from previous HPLC-MS methods for direct detection of glycidol esters or MCPD esters, this is the first HPLC-MS method used for the detection of free glycidol and total free MCPD in edible oil. Furthermore, this method can be potentially developed for glycidol or monochloropropane diol esters, which is similar to the current official methods adopted for indirect detection of these contaminants in different food matrices. Application of this detection method to real dietary supplements (fish oil and krill oil) revealed MCPD residues in fish oil (maximum detected: 32.78 ng·mL -1 ) and both MCPD (maximum detected: 2767.3 ng·mL -1 ) and glycidol (maximum detected: 22.2 ng·mL -1 ) in krill oil, emphasizing its effectiveness and accuracy for assessing contamination in these supplements.

SOCS (Suppressor of Cytokine Signalling) proteins are intracellular negative regulators that primarily modulate and inhibit cytokine-mediated signal transduction, playing a crucial role in immune homeostasis and related inflammatory diseases. SOCS act as inhibitors by regulating the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway, thereby intervening in the pathogenesis of inflammation and autoimmune diseases. Recent studies have also demonstrated their involvement in central immunity and neuroinflammation, showing a dual functionality. However, the specific mechanisms of SOCS in the central nervous system remain unclear. This review thoroughly elucidates the specific mechanisms linking the SOCS-JAK-STAT pathway with the inflammatory manifestations of neurodegenerative diseases. Based on this, it proposes the theory that SOCS proteins can regulate the JAK-STAT pathway and inhibit the occurrence of neuroinflammation. Additionally, this review explores in detail the current therapeutic landscape and potential of targeting SOCS in the brain via the JAK-STAT pathway for neuroinflammation, offering insights into potential targets for the treatment of neurodegenerative diseases., 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 The Authors. Published by Elsevier Inc. All rights reserved.)

Endothelial cells play an important role in the metabolism of adipose tissue (AT). This study aimed to analyze the changes that adipose tissue in AT endothelial cells undergo during the development of obesity, using single-nucleus RNA sequence (snRNA-seq). Mouse paraepididymal AT cells were subjected to snRNA-seq with the 10X Genomics platform. The cell types were then clustered using t-distributed stochastic neighbor embedding and unbiased computational informatics analyses. Protein-protein interactions network was established using the STRING database and visualized using Cytoscape. The dataset was subjected to differential gene enrichment analysis. In total, 21,333 cells acquired from 24 mouse paraepididymal AT samples were analyzed using snRNA-seq. This study identified 18 distinct clusters and annotated macrophages, fibroblasts, epithelial cells, T cells, endothelial cells, stem cells, neutrophil cells, and neutrophil cell types based on representative markers. Cluster 12 was defined as endothelial cells. The proportion of endothelial cells decreased with the development of obesity. Inflammatory factors, such as Vegfa and Prdm16 were upregulated in the medium obesity group but downregulated in the obesity group. Genes, such as Prox1, Erg, Flt4, Kdr, Flt1, and Pecam1 promoted the proliferation of AT endothelial cells and maintained the internal environment of AT. This study established a reference model and general framework for studying the mechanisms, biomarkers, and therapeutic targets of endothelial cell dysfunction-related diseases at the single-cell level., 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 Inc. All rights reserved.)

A 15-channel pressure filtration purification method was presented for high throughput sample preparation of aquatic products. A cost-effective device was constructed and melamine sponge was selected as the cleanup sorbent. Upon interfacing with HPLC-MS/MS, the analytical procedure demonstrated its suitability for quantifying 160 pesticides and veterinary drug residues in aquatic products such as fish, shrimp, and crab. The method achieved sample recoveries ranging from 61.3 to 124.9 %. The detection limits were established between 0.5 and 1.0 μg/kg, while the quantitation limits were confirmed to be within the range of 1.0-2.0 μg/kg. The method was applied to quantify the pesticide and veterinary drug residues in mostly consumed aquatic products from five coastal provinces in China. The results showed significant differences between different aquatic products in the concentrations of pesticide and veterinary drug residues, implying the necessity of supervision for the accurate determination of pesticides and veterinary drugs., 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.)

Polymer-based pure organic room-temperature phosphorescent materials have tremendous advantages in applications owing to their low cost, vast resources, and easy processability. However, designing polymer-based room-temperature phosphorescent materials with large Stokes shifts as key requirements in biocompatibility and environmental-friendly performance is still challenging. By generating charge transfer states as the gangplank from singlet excited states to triplet states in doped organic molecules, we find a host molecule (pyrrolidone) that affords charge transfer with doped guest molecules, and excellent polymer-based organic room-temperature phosphorescent materials can be easily fabricated when polymerizing the host molecule. By adding polyaromatic hydrocarbon molecules as electron-donor in polyvinylpyrrolidone, efficient intersystem crossing and tunable phosphorescent from green to near-infrared can be achieved, with maximum phosphorescence wavelength and lifetime up to 757 nm and 3850 ms, respectively. These doped polyvinylpyrrolidone materials have good photoactivation properties, recyclability, advanced data encryption, and anti-counterfeiting. This reported design strategy paves the way for the design of polyvinylpyrrolidone-based room-temperature phosphorescent materials., (© 2024. The Author(s).)

The presence of veterinary drug residues in aquatic products represents a significant challenge to food safety. The current detection methods, limited in both scope and sensitivity, underscore the urgent need for more advanced techniques. This research introduces a swift and potent screening technique using high-performance liquid chromatography-high-resolution mass spectrometry (HPLC-HRMS) and a refined QuEChERS protocol, allowing simultaneous qualitative and semi-quantitative analysis of 192 residues. A comprehensive database, employing full scan mode and data-dependent secondary mass spectroscopy, enhances screening accuracy. The method involves efficient extraction using 90% acetonitrile, dehydration with Na 2 SO 4 , and acetic acid, followed by cleanup using dispersive solid-phase extract sorbent primary secondary amine. It is suitable for samples with varying fat content, offering detection limits ranging from 0.5 to 10 μg/kg, high recovery rates (60-120%), and low relative standard deviations (<20%). Practical application has validated its effectiveness for multi-residue screening, marking a significant advancement in food safety evaluation., 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 Authors. Published by Elsevier Ltd.)

The carcinogenic and teratogenic risks of nitrofurazone (NFZ) led to its restriction in aquatic products. Semicarbazide (SEM), one of its metabolites, is a primary focus of modern monitoring techniques. However, the SEM residue in aquatic products is believed to be formed through endogenous mechanisms, especially for aquatic crustaceans. In this article, we will discuss the source of SEM, including its usage as an antibiotic in aquatic products (nitrofurazone), its production during food processing (azodicarbonamide and hypochlorite treatment), its occurrence naturally in the body, and its intake from the environment. SEM detection techniques were divided into three groups: derivatization, extraction/purification, and analytical methods. Applications based on liquid chromatography and its tandem mass spectrometry, immunoassay, and electrochemical methods were outlined, as were the use of various derivatives and their assisted derivatization, as well as extraction and purification techniques based on liquid-liquid extraction and solid-phase extraction. The difficulties of implementing SEM for nitrofurazone monitoring in aquatic products from crustaceans are also discussed. Possible new markers and methods for detecting them are discussed. Finally, the present research on monitoring illicit nitrofurazone usage through its metabolites is summarised, and potential problems that need to be overcome by continuing research are proposed with an eye toward giving references for future studies., 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 Authors.)

The follicle is an important unit for the synthesis of steroid hormones and the oocyte development and maturation in mammals. However, the effect of methionine supply on follicle development and its regulatory mechanism are still unclear. In the present study, we found that dietary methionine supplementation during the estrous cycle significantly increased the number of embryo implantation sites, as well as serum contents of a variety of amino acids and methionine metabolic enzymes in rats. Additionally, methionine supplementation markedly enhanced the expression of rat ovarian neutral amino acid transporters, DNA methyltransferases (DNMTs), and cystathionine gamma-lyase (CSE); meanwhile, it significantly increased the ovarian concentrations of the metabolite S -adenosylmethionine (SAM) and glutathione (GSH). In vitro data showed that methionine supply promotes rat follicle development through enhancing the expression of critical gene growth differentiation factor 9 and bone morphogenetic protein 15 . Furthermore, methionine enhanced the relative protein and mRNA expression of critical genes related to estrogen synthesis, ultimately increasing estrogen synthesis in primary ovarian granulosa cells. Taken together, our results suggested that methionine promoted follicular growth and estrogen synthesis in rats during the estrus cycle, which improved embryo implantation during early pregnancy. These findings provided a potential nutritional strategy to improve the reproductive performance of animals.

Moenomycin A, an antimicrobial growth promoter widely used as an additive in aquaculture feedstuffs, has been restricted for use in the European Union and China due to its potential risk of promoting resistant strains of pathogenic bacteria and causing residues in aquatic animal products. Although methods for analyzing moenomycin A in feedstuffs have been developed, no established method exists for aquatic matrices. In this study, we present, for the first time, a sensitive and validated high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of moenomycin A in aquatic animal products. Samples were extracted using methanol and purified with the QuEChERS method employing C18 sorbent. The aliquot was dried under a nitrogen stream, reconstituted with methanol-water solvent, and analyzed by HPLC-MS/MS. The developed method exhibited good linearity (r 2  > 0.995) over a wide concentration range (1-100 μg/L) and a low limit of detection (1 µg/kg). Average recoveries ranged between 70 and 110% at spiked concentrations of 1, 50, and 100 μg/kg, with associated intra- and inter-day relative standard deviations of 1.25 to 7.32% (n = 6) and 2.91 to 10.08% (n = 3), for different representative aquatic animal production, respectively. To the best of our knowledge, this is the first reported HPLC-MS/MS method for the quantification of moenomycin A in aquatic animal products. The new approach was effectively employed in the analysis of moenomycin A across various aquatic samples., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)

A novel, simplified derivatization method and a rapid sample preparation process using carbon yarn as a sorbent for the determination of 3-chloropropane-1,2-diol (3-MCPD) in soy sauce via high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed. 3-MCPD was first enriched and purified with carbon yarn and then eluted with a methanol-water solution. Subsequently, the analyte underwent derivatization with p -(dimethylamino)-phenol for sensitive detection via HPLC-MS/MS. The limit of detection and the limit of quantitation for 3-MCPD were validated to be 0.5 and 1.0 μg/kg, respectively. Spiking experiments showed recoveries between 83 and 94%, with a relative standard deviation of ≤10%. The method was further validated with a certified reference material. Furthermore, 11 real soy sauce samples from local markets were tested by using this method. These results reveal the widespread 3-MCPD contamination. Consequently, this study offers a preferable alternative for the sensitive, accurate, and precise determination of 3-MCPD in soy sauce.

A modified QuEChERS method was developed to determine multi-class pesticide and veterinary residues in aquatic products. Chitosan microspheres were conveniently synthesized and utilized as the cleanup adsorbent in the QuEChERS procedure, showcasing rapid filtration one-step pretreatment ability for the determination of drug multi-residues in aquatic products. Compared to conventional synthetic sorbents, chitosan microspheres not only have good purification performance, but also have renewable and degradable properties. This novel sorbent worked well in the simultaneous determination of 95 pesticides and veterinary drug residues in aquatic products after being combined with an improved one-step vortex oscillating cleanup method. We achieved recoveries ranging from 64.0% to 115.9% for target drugs in shrimp and fish matrix. The limits of detection and quantification were 0.5-1.0 and 1.0-2.0 μg kg -1 , respectively. Notably, hydrocortisone was detected with considerable frequency and concentration in the tested samples, underscoring the necessity for stringent monitoring of this compound in aquatic products., 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 Ltd.)

Microcin C7 (McC) as a viable form of antimicrobial has gained substantial attention due to its distinctive antimicrobial activity, by targeting aspartyl tRNA synthetase. McC can be a potential solution against pathogenic microbial infections in the postantibiotic era. However, considering that degradation by digestive enzymes can disrupt the function of this peptide in the gastrointestinal tract, in this study, we attempt to design McC variants to overcome several barriers that may affect its stability and biological activity. The mccA gene encoding the McC peptide precursor was mutated and 12 new McC variants with trypsin resistance were found. The Yej + rimL - strain was used as an indicator to determine the minimum inhibitory concentrations (MICs). The results showed that three variants, including R2A, R2T and R2Q, among 12 variants formed by the replacement of the second arginine of the McC peptide with different amino acids, were resistant to trypsin and had an outstanding antimicrobial ability, with MIC values of 12.5, 25, and 25 μg/mL, respectively. Taken together, our findings show that the engineering of the site-directed mutagenesis of McC significantly enhances McC trypsin resistance and maintains a great antimicrobial activity.

In this study, a novel filter-press cleanup column was developed as a single-step cleanup approach for the rapid screening and quantification of 112 veterinary drugs in fish samples. Fish muscle samples were extracted with acetonitrile and ethyl acetate, sequentially. After concentration and reconstitution, N -propylethylenediamine (PSA) sorbent, packed in filter-press column, allows rapid single-step cleanup operation, while UHPLC-Q-Orbitrap-HRMS provides high-precision mass information in multi-residue screening. Under optimum settings, the detection and quantification limits were validated at 0.5 and 2.0 μg·kg -1 , for all analytes, respectively. The ranges of recoveries were from 35.3 to 138.4%. Most of these target analytes (82%) could be measured with recoveries between 60 and 130%, and intra-day RSDs ranging from 1.9 to 26.1%. This method was further applied to evaluate the residual of veterinary drugs in fish samples from four cities in China, and results demonstrated its practicability for multi-residue monitoring veterinary residues for food safety administration., 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., (© 2023 The Authors.)

Notably, the intermolecular charge transfer between pyrene (Py) and benzophonenes (BPs) can significantly enhance the quantum yield of the triplet state of Py, which will convert Py from a fluorescence molecule to a phosphorescence molecule. The intermolecular charge transfer is confirmed by steady-state and time-resolved spectroscopy and theoretical study. Based on these foundations, Py is doped into BPs systems and a large Stokes-shift organic room temperature phosphorescence (ORTP) is observed. By using different benzophenone derivatives, a series of host-guest ORTP materials with different luminescent properties adjusted by intermolecular charge transfer features are developed. Fortunately, these host-guest ORTP systems from benzophenone derivatives and pyrene are readily fabricated, and the red gradient color lasting as long as 3 s is observed after removing UV excitation. This host-guest charge transfer strategy plays an important role in the mechanism of the luminous type shift. Our strategy paves the way to design ORTP materials conveniently and apply these materials in encryption and temperature alarm device.

A single-layer graphene metamaterial consisting of a horizontal graphene strip, four vertical graphene strips, and two graphene rings is proposed to realize tunable multi-plasma-induced transparency (MPIT) by the coupled mode theory and the finite-difference time-domain method. A switch with three modulation modes is realized by dynamically adjusting the Fermi level of graphene. Moreover, the effect of symmetry breaking on MPIT is investigated by controlling the geometric parameters of graphene metamaterials. Triple-PIT, dual-PIT, single-PIT can be transformed into each other. The proposed structure and results provide guidance for applications such as designing photoelectric switches and modulators.

CD34 + cells improve the perfusion and function of ischemic limbs in humans and mice. However, there is no direct evidence of the differentiation potential and functional role of these cells in the ischemic muscle microenvironment. Here, we combined the single-cell RNA sequencing and genetic lineage tracing technology, then provided exact single-cell atlases of normal and ischemic limb tissues in human and mouse, and consequently found that bone marrow (BM)-derived macrophages with antigen-presenting function migrated to the ischemic site, while resident macrophages underwent apoptosis. The macrophage oncostatin M (OSM) regulatory pathway was specifically turned on by ischemia. Simultaneously, BM CD34 + -derived proregenerative fibroblasts were recruited to the ischemia niche, where they received macrophage-released OSM and promoted angiopoietin-like protein-associated angiogenesis. These findings provided mechanisms on the cellular events and cell-cell communications during tissue ischemia and regeneration and provided evidence that CD34 + cells serve as fibroblast progenitors promoting tissue regeneration.

Ethylene biosynthesis in apple (Malus domestica) fruit can be suppressed by calcium ions (Ca2+) during storage; however, the underlying mechanisms are unclear. In this study, we identified the apple transcription factor MCM1-AGAMOUS-DEFICIENS-SRF5 (MdMADS5), which functions as a transcriptional activator of the ethylene biosynthesis-related gene 1-AMINOCYCLOPROPANE-1-CARBOXYLIC ACID SYNTHASE1 (MdACS1), a partner of the calcium sensor CALCIUM-DEPENDENT PROTEIN KINASES7 (MdCDPK7). Ca2+ promoted the MdCDPK7-mediated phosphorylation of MdMADS5, which resulted in the degradation of MdMADS5 via the 26S proteasome pathway. MdCDPK7 also phosphorylated 1-AMINOCYCLOPROPANE-1-CARBOXYLIC ACID OXIDASE1 (MdACO1), the key enzyme in ethylene biosynthesis, leading to MdACO1 degradation and inhibition of ethylene biosynthesis. Our results reveal that Ca2+/MdCDPK7-MdMADS5 and Ca2+/MdCDPK7-MdACO1 are involved in Ca2+-suppressed ethylene biosynthesis, which delays apple fruit ripening. These findings provide insights into fruit ripening, which may lead to the development of strategies for extending the shelf life of fruit., Competing Interests: Conflict of interest statement. The authors declare no conflict of interest., (© American Society of Plant Biologists 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Pesticide residues in aquatic products are of great concern due to the risk of environmental transmission and their extensive use in aquaculture. In our work, a quick screening approach was developed for the qualitative and semi-quantitative screening of 87 pesticide residues in aquatic products. The sample preparation was investigated, including extract solvent, extract methods, buffer salts, lipid removal, cleanup materials and filter membranes for aquatic products. Samples were extracted using a modified QuEChERS procedure, and two clean-up procedures were developed for UHPLC-Q/Orbitrap MS analysis based on the fat content of the aquatic products. The screening detection limits for all studied pesticides were distributed between 1 and 500 μg/kg in the three representative matrices. Seventy-one pesticides could be analyzed with a screening limit between 1 and 25 μg/kg in grass carp and crayfish, sixty-one pesticides could be screened for limits between 1 and 50 μg/kg in crab. The accuracy results showed that recoveries ranged from 50 to 120% for 60, 56 and 52 pesticides at medium-level for grass carp, crayfish and crab, respectively. At high spiking levels, 74, 65 and 59 pesticides were recovered within the range of 50-120% for the three matrices, respectively. The relative standard deviations of most compounds in different matrices were less than 20%. With this method, the local farmed aquatic products were tested for pesticide residues. In these samples, ethoxyquinoline, prometryn and phoxim were frequently detected. The majority of these confirmed compounds did not exceed 2.00 μg/kg. A grass carp with trichlorfon at 4.87 μg/kg and two carps with ethoxyquinoline at 200 µg/kg were detected, indicating the potential dietary risk.

Extensive and high residue variations in enrofloxacin (ENR) exist in different aquatic products. A novel quantitative method for measuring ENR using high-performance liquid chromatography-tandem mass spectrometry was developed employing enrofloxacin- d 5 (ENR- d 5 ) and enrofloxacin- d 3 (ENR- d 3 ) as isotope surrogates. This reduced the deviation of detected values, which results from the overpass of the linear range and/or the large difference in the residue between the isotope standard and ENR, from the actual content. Furthermore, high residue levels of ENR can be directly diluted and re-calibrated by the corresponding curve with the addition of high levels of another internal surrogate without repeated sample preparation, avoiding the overflow of the instrument response. The validation results demonstrated that the method can simultaneously determine ENR residues from MQL (2 µg/kg) to 5000 × MQL (method quantification limit) with recoveries between 97.1 and 106%, and intra-precision of no more than 2.14%. This method realized a wide linear calibration range with dual deuterated isomers, which has not been previously reported in the literature. The developed method was successfully applied to the analysis of ENR in different aquatic products, with ENR residue levels varying from 108 to 4340 μg/kg and an interval of precision in the range of 0.175~6.72%. These results demonstrate that batch samples with a high variation in ENR residues (over the linear range with a single isotope standard) can be detected by the dual isotope surrogates method in a single sample preparation process.

One-carbon metabolism is a key metabolic network that integrates nutritional signals with embryonic development. However, the response of one-carbon metabolism to methionine status and the regulatory mechanisms are poorly understood. Herein, we found that methionine supplementation during pregnancy significantly increased fetal number and average fetal weight. In addition, methionine modulated one-carbon metabolism primarily through 2 metabolic enzymes, cystathionine β-synthase (CBS) and methionine adenosyltransferase 2A (MAT2A), which were significantly increased in fetal liver tissues and porcine trophoblast (pTr) cells in response to proper methionine supplementation. CBS and MAT2A overexpression enhanced the DNA synthesis in pTr cells. More importantly, we identified a transcription factor, DNA damage-inducible transcript 3 (DDIT3), that was the primary regulator of CBS and MAT2A, which bound directly to promoters and negatively regulated the expression of CBS and MAT2A. Taken together, our findings identified that DDIT3 targeting CBS and MAT2A was a novel regulatory pathway that mediated cellular one-carbon metabolism in response to methionine signal and provided promising targets to improve pregnancy health., Competing Interests: Declaration of Competing Interests The authors declare that there are no conflicts of interest., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Prometryn, a widely used triazine herbicide in agriculture and aquaculture, has been commonly detected in marine environments, but its effects on the marine copepod are unknown. In this study, marine copepod Tigriopus japonicus was chronically exposed to environmentally relevant concentrations of prometryn to investigate its impacts and potential mechanism of action. The results showed that 0.5, 5, and 50 μg/L prometryn delayed the first spawning time and hatching time, reduced the fecundity, and inhibited the population growth rate. Moreover, exposure to 0.5, 5 and 50 μg/L prometryn decreased food ingestion, the content of C and N elements, nutrient accumulation and body size, but increased the content of 20-hydroxyecdysone (20E). Transcriptome analysis showed that 50 μg/L prometryn down-regulated 1431 genes, which were mainly enriched in lysosome pathway and chitin binding and cuticle construction process. The results of qRT-PCR showed that the expression of key genes involved in juvenile hormone synthesis and chitin metabolic pathways were also inhibited after prometryn exposure. Molecular docking revealed that prometryn could bind to ecdysone receptor (EcR) and UDP-N-acetylglucosamine pyrophosphorylase (UAP), components of the ecdysteroid nuclear receptor complex. Therefore, environmental relevant prometryn delayed the molting and development of T. japonicus by disrupting the ecdysone signal pathway and chitin metabolic pathway through binding to EcR and UAP. This study provides new insights into toxic effects and molecular mechanisms of prometryn on marine copepods., 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.)

Malachite green (MG) is a synthetic poisonous organic compound that has been banned in many countries as a veterinary drug for aquaculture. An efficient, fast and sensitive method is urgently needed for monitoring the illegal use of malachite green (MG) in aquaculture. In this study, a novel ratiometric fluorescence immunoassay was established. Nitrogen-doped carbon quantum dots were used as ratiometric fluorescent probes with a fluorescence peak at 450 nm. Horseradish peroxidase was employed to convert o-phenylenediamine to 2,3-diaminophenazine, with a new fluorescence peak at 580 nm and a strong absorption at 420 nm. The inner filter effect between N-CQD fluorescence and DAP absorption was identified. It allows for the ratiometric detection of MG using a fluorescent immunoassay. The results demonstrated a linear ratiometric fluorescence response for MG between 0.1 and 12.8 ng·mL -1 . The limit of detection of this method was verified to be 0.097 μg·kg -1 with recoveries ranging from 81.88 to 108%, and the relative standard deviations were below 3%. Furthermore, this method exhibited acceptable consistency with the LC-MS/MS results when applied for MG screening in real samples. These results demonstrated a promising application of this novel ratiometric fluorescence immunoassay for MG screening with the merits of rapid detection, simple sample preparation, and stable signal readout. It can be an alternative to other traditional methods if there are difficulties in the availability of expensive instruments, and achieve comparable results or even more sensitivity than other reported methods.

This paper proposes a novel and perfect absorber based on patterned graphene and vanadium dioxide hybrid metamaterial, which can not only achieve wide-band perfect absorption and dual-channel absorption in the terahertz band, but also realize their conversion by adjusting the temperature to control the metallic or insulating phase of VO 2 . Firstly, the absorption spectrum of the proposed structure is analyzed without graphene, where the absorption can reach as high as 100% at one frequency point (f = 5.956 THz) when VO 2 is in the metal phase. What merits attention is that the addition of graphene above the structure enhances the almost 100% absorption from one frequency point (f = 5.956 THz) to a wide frequency band, in which the broadband width records 1.683 THz. Secondly, when VO 2 is the insulating phase, the absorption of the metamaterial structure with graphene outperforms better, and two high absorption peaks are formed, logging 100% and 90.7% at f 3  = 5.545 THz and f 4  = 7.684 THz, respectively. Lastly, the adjustment of the Fermi level of graphene from 0.8 eV to 1.1 eV incurs an obvious blueshift of the absorption spectra, where an asynchronous optical switch can be achieved at fK 1  = 5.782 THz and fK 2  = 6.898 THz. Besides, the absorber exhibits polarization sensitivity at f 3  = 5.545 THz, and polarization insensitivity at f 4  = 7.684 THz with the shift in the polarization angle of incident light from 0° to 90°. Accordingly, this paper gives insights into the new method that increases the high absorption width, as well as the great potential in the multifunctional modulator.

Background: The approach to matching appropriate carbohydrates alongside free amino acids to achieve optimal muscle growth remains unclear., Objectives: We investigated whether the consumption of a diet containing rapidly digested carbohydrate and free amino acids can enhance intestinal absorption and muscular uptake of amino acids in pigs., Method: Twelve barrows (28 kg; 11 wk old) with catheters installed in the portal vein, mesenteric vein, femoral artery, and femoral vein were randomly assigned to consume 1 of 2 free amino acid-enriched diets (3.34%) containing rapidly [waxy corn starch (WCS)] or slowly [pea starch (PS)] digested carbohydrate for 27 d. Blood was collected to determine the fluxes of plasma glucose and amino acids across the portal vein and the hindlimb muscle. Dietary in vitro carbohydrate digestive rates were also determined. Data were analyzed using repeated-measures (time × group) ANOVA., Results: Carbohydrate in vitro cumulative digestibility at 30 and 240 min was 69.00% and 95.25% for WCS and 23.25% and 81.15% for PS, respectively. The animal experiment presented WCS increased individual amino acids (lysine, 0.67 compared with 0.53 mmol/min; threonine, 0.40 compared with 0.29 mmol/min; isoleucine, 0.33 compared with 0.22 mmol/min; glutamate, 0.51 compared with 0.35 mmol/min; and proline, 0.51 compared with 0.27 mmol/min), essential amino acid (EAA; 3.26 compared with 2.65 mmol/min), and branched-chain amino acid (BCAA; 0.86 compared with 0.65 mmol/min) fluxes across the portal vein during 8 h postprandial, as well as individual amino acids (isoleucine, 0.08 compared with 0.02 mmol/min; leucine, 0.06 compared with 0.02 mmol/min; and glutamine, 0.44 compared with 0.25 mmol/min), EAA (0.50 compared with 0.21 mmol/min), and BCAA (0.17 compared with 0.06 mmol/min) net fluxes across the hindlimb muscle during 8 h postprandial compared with PS (P < 0.05)., Conclusions: A diet containing rapidly digested carbohydrate and free amino acids can promote intestinal absorption and net fluxes across hindlimb muscle of amino acids in pigs., (Copyright © 2022 American Society for Nutrition.)

Double narrowband induced perfect absorption in the terahertz region is achieved in a graphene-dielectric-gold hybrid metamaterial, whose physical mechanism is analyzed using the coupled-mode theory (CMT), which agreed well with the finite-difference time-domain (FDTD) simulation. This study found that the Fermi level of graphene can be adjusted to improve the absorptivity when the refractive index (RI) n d of the chosen dielectric cannot achieve a good absorption effect. In addition, the blue shift of absorption spectrum can be used in the design of dual-frequency electro-optical switches, of which the modulation degree of amplitude (MDA) can reach as high as 94.05% and 93.41%, indicating that this is a very promising electro-optical switch. Most significantly, the RI sensing performance is investigated, which shows an ultra-high absorption sensitivity S A  = 4.4°/RIU, wavelength sensitivity S λ  = 9.8°/RIU, and phase shift sensitivity S φ  = 2691°/RIU. At last, an interesting finding is that the two peaks (R1 and R2) of plasmon-induced absorption (PIA) show different polarization characteristics (insensitive or sensitive) to the incident light angle; this polarization-sensitive is particularly important for the PIT/PIA-based optical polarizers. Undoubtedly, this paper is of great significance to the research and design of terahertz photonic devices and sensors., (© 2022. The Author(s).)

The occurrence of antibiotics and potential health risk of 300 cultured fish samples from 19 provinces in China were investigated. The levels of 28 antibiotics (15 fluoroquinolones, 4 tetracyclines, 8 macrolides and rifampin) in 8 fish species were measured through liquid chromatography electrospray tandem mass spectrometry. As a result, 10 antibiotics were detected with an overall detection frequency of 24.3%, and the individual detection frequency of antibiotics ranged from 0.33 to 16.7%. The extremely high concentrations (above 100 µg/kg) of doxycycline and erythromycin were found in the samples. Antibiotics with high detection frequency was noticed in largemouth bass (41.2%), followed by snakehead (34.4%) and bream (31.2%). Specifically, Heilongjiang, Xinjiang, Qinghai and Gansu presented high detection frequency values of more than 60%. Moreover, the highest mean concentration was observed in Shandong, and the concentration covered from 34.8 µg/kg to 410 µg/kg. Despite the high detection frequency and levels of antibiotics were found in samples, ingestion of cultured fish was not significantly related to human health risks in China, according to the calculated estimated daily intakes and hazard quotients. These results provided us the actual levels of antibiotics in cultured fish and human health risk assessment of consuming fishery products., Competing Interests: 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 © 2022 Tang, Lou, Yang, Tian, Wang and Huang.)

Background: Reduced nutrient digestibility due to low-protein (LP) diets occurring in the foregut or hindgut of pigs remains unclear. Methods: Growing barrows (21.7 ± 1.7 kg) were allotted into LP and high-protein (HP) diet treatments. Ileal digesta and feces were collected for in vitro cross-fermentation and microbial sequencing, and cross-feeding assessed nutrient digestibility. Results: No difference in foregut digesta flora and nutrient digestibility between treatments was observed. LP diet caused decreased total tract digestibility of dry matter (DM), organic matter (OM), gross energy (GE), neutral detergent fiber (NDF), and acid detergent fiber (ADF) compared with the HP diet (p < 0.05). The fermentation broth from LP diet-fed pigs induced less full fermentation digestion of DM, OM, crude protein, and GE than HP broth (p < 0.05). Additionally, LP broth fermentation presented lower fermentation gas and short-chain fatty acids (SCFAs) generation than HP group (p < 0.05). This situation above may be related to decreased abundances of Lachnospiraceae, Eubacterium&#95;eligens&#95;group, Roseburia, and Ruminococcaceae&#95;UCG-009, which can efficiently ferment nutrients to produce SCFA. Conclusions: Change in the flora caused compromise in hindgut microbial fermentation digestion leads to decreased total tract nutrient digestibility in pigs fed an LP diet.

Objective: The study was aimed to investigate the rules of postprandial changes in intestine digesta dry matter (DM) flow and amino acid digestibility of growing pigs fed low-protein (LP) diets made of different starch., Methods: Eight barrows (28.8±2.1 kg) with a T-cannula at the distal ileum were randomly allotted to an 8×3 Youden square design. Treatments included: waxy corn starch LP (WLP); corn starch LP (CLP) and pea starch LP (PLP). Diets were given at 08:00 and 20:00. Digesta samples were collected in six 2-h stages from 08:00 to 20:00., Results: The Cr concentrations of ileal digesta increased and then decreased in WLP and CLP, while increased continuously in PLP as time passed after postprandial (p<0.05). Higher average Cr concentrations (0.78% and 0.84% vs 0.70%; p<0.05) and lower average DM flow (181.1 g/kg and 166.3 g/kg vs 240.3 g/kg; p<0.001) were observed in WLP and CLP, compared with PLP. The apparent ileal digestibility coefficient of most amino acids in WLP and CLP increased compared with that in PLP. No difference in lysine or methionine digestibility was observed. When digesta were collected in 2-h periods, the apparent ileal digestibility coefficient of amino acids did not change over time. When digesta was collected in 4-h periods from 16:00 to 20:00 and 6-h periods from 14:00 to 20:00 (p<0.05), WLP and CLP showed markedly higher amino acid digestibility than PLP., Conclusion: High-amylose slowly digested starch can increase the DM flow at the distal ileum and reduce the apparent ileal digestibility coefficient of amino acids of pigs fed LP diets. Compared with waxy corn starch and corn starch, pea starch reduced the digestibility of amino acids in digesta collected after 4 h postprandial.

Background: Protein releases amino acids faster than starch releases glucose in digestive tract of pigs fed low-protein (LP) diets. Poor synchronization of dietary glucose and amino acids supply leads to compromised nitrogen efficiency. Dietary starch patterns modulation may improve this situation., Methods: Growing barrows (29.7 ± 2.0 kg) were randomly allotted into 5 dietary treatments with LP diets consisting of different purified starches. Treatments included: waxy corn starch (W LP), corn starch + waxy corn starch (C + W LP), corn starch (C LP), pea starch + waxy corn starch (P + W LP) and pea starch (P LP). In the experiment, growth performance, protein deposition, nutrient metabolism, and fecal microbial community of pigs were investigated. In vitro starch digestion was used for predicting the in vivo glucose response., Results: Dietary starch in vitro glucose release profile was determined by starch source and the ratio of amylopectin and amylose. C + W LP treatment showed decreased total nitrogen excretion and plasma citrulline concentration and improved plasma leptin concentration among treatments (P < 0.05). Besides, the highest nitrogen apparent biological value, whole-body protein deposition and growth performance and lowest urinary nitrogen excretion were also observed in C + W LP treatment. Compared with the other groups, C + W LP and C LP showed increased plasma pyruvate, IGF-1, and lipase concentrations (P < 0.05). The W LP group presented dramatically increased plasma alanine and urea nitrogen concentration and decreased aldolase and leptin concentrations (P < 0.05). Dietary starch patterns did not make an impact on bacterial richness and diversity, but changed the taxonomic and functional structures of the microbial communities. Microbial protein fermentation product (isobutyrate and isovalerate) presented increased in P LP treatments compared with the other treatments (P < 0.05)., Conclusions: Dietary starch patterns modulation can regulate dietary glucose release profile, nutrient metabolism, protein turnover, and fecal microbial fermentation in pigs. The optimal dietary glucose release profile effectively strengthened whole-body protein deposition and improve nitrogen efficiency and growth performance in growing pigs fed LP diets., (© 2022. The Author(s).)

A simple monolayer graphene metamaterial based on silicon/silica substrates is proposed, and typical triple-plasmon-induced transparency (PIT) is realized in the terahertz band. The physical mechanism is analyzed by coupled mode theory (CMT), and the results of CMT agree well with the finite-difference time-domain simulation. A multimode electro-optical switch can be designed by dynamic tuning, and the modulation degrees of its resonant frequencies are 84.0%, 87.3%, 83.0%, 88.1%, and 76.7%. In addition, triple-PIT gradually degenerates into dual-PIT with a decrease in the length of one bright mode. Interestingly, the group index can reach 770 at E f =0.8 e V , which shows that it can be designed as a slow light device with extraordinary ability. Therefore, the results of this paper are of great significance to the research and design of electro-optical switches and slow light devices in the terahertz band.

T lymphocyte and macrophage infiltration in the aortic wall is critical for abdominal aortic aneurysm (AAA). However, how T lymphocytes interact with macrophages in the pathogenesis of AAA remains largely uncharacterized. In an elastase-induced murine AAA model, we first found that the expression of pyruvate kinase muscle isozyme 2 (PKM2), the last rate-limiting enzyme in glycolysis, was increased in infiltrated T lymphocytes of vascular lesions. T lymphocyte-specific PKM2 deficiency in mice (LckCrePKM2 fl/fl ) or intraperitoneal administration of the sphingomyelinase inhibitor GW4869 caused a significant attenuation of the elastase-increased aortic diameter, AAA incidence, elastic fiber disruption, matrix metalloproteinases (MMPs) expression, and macrophage infiltration in the vascular adventitia compared with those in PKM2 fl/fl mice. Mechanistically, extracellular vesicles (EVs) derived from PKM2-activated T lymphocytes elevated macrophage iron accumulation, lipid peroxidation, and migration in vitro, while macrophages treated with EVs from PKM2-null T lymphocytes or pretreated with the lipid peroxidation inhibitors ferrostatin-1 (Fer-1), liproxstatin-1 (Lip-1), or the iron chelating agent deferoxamine mesylate (DFOM) reversed these effects. In vascular lesions of elastase-induced LckCrePKM2 fl/fl mice with AAA, the oxidant system weakened, with downregulated 4-hydroxynonenal (4-HNE) levels and strengthened antioxidant defense systems with upregulated glutathione peroxidase 4 (GPX4) and cystine/glutamate antiporter solute carrier family 7 member 11 (Slc7a11) expressions in macrophages. High-throughput metabolomics showed that EVs derived from PKM2-activated T lymphocytes contained increased levels of polyunsaturated fatty acid (PUFA)-containing phospholipids, which may provide abundant substrates for lipid peroxidation in target macrophages. More importantly, upregulated T lymphocyte PKM2 expression was also found in clinical AAA subjects, and EVs isolated from AAA patient plasma enhanced macrophage iron accumulation, lipid peroxidation, and migration ex vivo. Therefore, from cell-cell crosstalk and metabolic perspectives, the present study shows that PKM2-activated T lymphocyte-derived EVs may drive AAA progression by promoting macrophage redox imbalance and migration, and targeting the T lymphocyte-EV-macrophage axis may be a potential strategy for early warning and treating AAA., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Eugenol is the most commonly used plant anesthetic to relieve the stressors during various aquaculture procedures. This study aims to investigate the pharmacokinetics of eugenol in Pacific white shrimp by immersion baths in a simulated transportation., Results: The pharmacokinetics of eugenol were firstly investigated in Pacific white shrimp by immersion baths of 300 mg L - 1 eugenol over 5 min (Treatment 1), 10 mg L - 1 eugenol during 24 h (Treatment 2) and a sequential immersion administration (Treatment 3). Concentrations of eugenol in hemolymph, hepatopancreas, and muscle were determined using Gas chromatography-tandem mass spectrometry (GC-MS/MS). After immersion bath of Treatment 1, the elimination half-life (t 1/2z ) values are 1.3 h and 11 h for hepatopancreas and muscles, indicating the rapid absorption and elimination of eugenol in shrimp. Under the Treatment 2 administration, the eugenol peak concentration is 6527.9 μg/kg in muscle, followed by 402.8 μg/kg in hepatopancreas, with the lowest concentration of 37.9 μg/L in hemolymph. Area under the curve (AUC 0-∞ ) values lie in the order of muscle > hepatopancreas > hemolymph, suggesting that eugenol tends to accumulate in muscle by the immersion administration. Moreover, the average residence time (MRT 0-∞ ) values of 38.6, 23.0 and 115.3 h for hemolymph, hepatopancreas and muscle are achieved, which may indicate that hepatopancreas is the main organ for elimination of eugenol. After combining the conditions in a sequential bath immersion of eugenol (Treatment 3), the maximum concentration (C max ) values of eugenol are higher than those achieved in Treatment 2, indicating that accumulation of eugenol happened in haemolymph, hepatopancreas and muscle. In addition, the corresponding t 1/2z values are 4.7, 14.9 and 47.6 h, respectively, suggesting the faster elimination from the tissues following sequential administration. After the immersion bath, eugenol concentrations in muscle of Pacific white shrimp are lower than 2.5 mg/kg at 2 h, 48 h and 24.5 h in Treatment 1 ~ 3., Conclusions: A withdrawal period of 2 h, 48 h and 24.5 h following a 300 mg L - 1 of eugenol over a 5-min, 10 mg L - 1 eugenol concentration during a 24-h and combined conditions in a sequential immersion bath were suggested., (© 2022. The Author(s).)

A terahertz-band metamaterial composed of multilayer patterned graphene is proposed and triple plasmon-induced transparency is excited by coupling three bright modes with one dark mode. The Lorentz curve calculated by the coupled-mode theory agrees well with the finite-difference time-domain results. Dynamic tuning is investigated by changing the Fermi level. Multimode electro-optics switching can be designed and achieved, and the amplitude modulations of four resonance frequencies are 94.3%, 92.8%, 90.7%, and 93%, respectively, which can realize the design of synchronous and asynchronous electro-optics switches. It is hoped that these results can provide theoretical support and guidance for the future design and application of photonic and optoelectronic devices.

Multidrug resistant (MDR) bacterial infection has emerged, raising concerns about untreatable infections, and posing the highest health risks. Antimicrobial peptides (AMPs) are thought to be the best remedy for this problem. Here, we showed biosynthetic microcin J25 (MccJ25) exhibited excellent bactericidal activity against standard and clinically relevant veterinary MDR strains with high stability, no cytotoxicity, and no increase in drug resistance. Analysis of antimicrobial mechanism possessed by sensitive enterotoxigenic Escherichia coli (ETEC) based on electron microscopy and Sytox Green methods was carried out. Results showed excellent activity against ETEC was due to permeabilizing bacterial membranes and strong affinity. MccJ25 exhibited high endotoxin-neutralizing activity in both in vivo and in vitro environments, and mice exposed to lipopolysaccharide (LPS) showed decreased plasma LPS levels and improved survival after administration of MccJ25. In an LPS-treated mouse septicemia model, MccJ25 treatment significantly alleviated inflammatory responses by inhibiting proinflammatory factor secretion and expression. In a mouse E. coli infection model, administration of MccJ25 effectively improved host defense against clinically source cocktail of multidrug-resistant E. coli strains induced intestinal inflammation and bacteria dissemination. Results of studies on anti-inflammatory mechanisms showed that MccJ25 downregulated nuclear factor kappa B kinase and mitogen-activated protein kinase, thereby reducing the production of toll-like receptor 4, myeloid differentiation factor 88 and decreasing the key proinflammatory cytokines. These findings clarify MccJ25 may be an ideal antibacterial/antiendotoxic drug candidate that has the potential to further guide the development of anti-inflammatory and/or antimicrobial agents in the war against MDR bacterial infection., Competing Interests: 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. The reviewer XW declared a shared affiliation, with no collaboration, with the authors to the handling editor at the time of the review., (Copyright © 2022 Yu, Shang, Yang, Dai, Zeng and Qiao.)

Microplastics (MPs) are considered to exacerbate the impacts of hydrophobic organic pollutants on aquatic organisms through the carrier function, but whether MPs affect the transgenerational toxicity of pollutants is unclear. This study exposed adult female marine medaka to phenanthrene (Phe)-adsorbed MPs for 60 days to investigate the effects of MPs on the bioaccumulation, reproductive and transgenerational toxicity of Phe. Compared to Phe alone, co-exposure to Phe and 200 μg/L MPs significantly increased Phe bioaccumulation in the intestines and ovaries. Phe alone and Phe combined with MPs disrupted the regulation of the hypothalamus-pituitary-gonadal axis, and reduced vitellogenin levels and reproductive capacity of female fish. In particular, co-exposure to 200 μg/L MPs and Phe increased the rate of follicular atresia, inhibited ovarian maturity, and aggravated reproductive toxicity. Notably, maternal uptake of Phe could be transferred to the offspring, and embryonic accumulation increased with the concentrations of MPs. Moreover, MPs aggravated Phe-induced bradycardia in embryos, suggesting that MPs exacerbated the transgenerational toxicity of Phe. These findings reveal that the growing number of MPs in the ocean might amplify the adverse effects of organic pollutants on the health and population stability of marine fishes, and this problem merits more attention., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A green and facile hydrothermal synthesis approach is proposed for the preparation of nitrogen-doped carbon quantum dots (N-CQDs) with wolfberry. These N-CQDs were developed as a highly sensitive fluorescent 'on-off-on' switch sensor for the sensing of Fe 3+ and l-ascorbic acid (AA). The N-CQDs displayed superior fluorescence characteristics of CQDs with a quantum yield up to 22%. The N-CQDs were demonstrated to selectively react with Fe 3+ , leading to fluorescence quenching effect, which was successfully used for the detection of Fe 3+ with a limit of detection at 3 μmoL•L -1 . The addition of AA is supposed to repair the surface defects, and result in the fluorescence recovery. Based on this effect, the strategy of 'on-off-on' detection of AA was established with a limit of detection at 1.8 μmoL•L -1 . Furthermore, the practical application of the detection of Fe 3+ lake water and AA in medical tablet was demonstrated, promising an effective and efficient 'on-off-on' nanosensor with low-cost, green synthesis for Fe 3+ and AA detection., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

A method for the determination of 8 biogenic amines in aquatic products and their derived products was established by HPLC-MS/MS without derivatization. The samples were extracted by 5% perchloric acid solution. N-hexane was used to clean the extract. The analytes were separated by a column of ACQUITY UPLC HSS T3 (100 mm × 2.1 mm, 1.8 µm), and gradient eluted with a mixed solution of (0.5% formic acid) and acetonitrile. Good linearity was obtained with correlation coefficients (R 2 ) >0.99. This method achieved higher sensitivity (from 0.1 mg/kg for tyramine, 2-phenylethylamine and tryptamine to 1.0 mg/kg for spermidine, spermine, cadaverin, histamine and putrescine). The average recoveries were demonstrated in the range of 70.9%-113.1%, with relative standard deviations (RSDs) from 0.33% to 10.81%. This method was suitable for the detection of BAs in aquatic products and their products., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

With the increasing maternal age and the use of assisted reproductive technology in various countries worldwide, the influence of epigenetic modification on embryonic development is increasingly notable and prominent. Epigenetic modification disorders caused by various nutritional imbalance would cause embryonic development abnormalities and even have an indelible impact on health in adulthood. In this scoping review, we summarize the main epigenetic modifications in mammals and the synergies among different epigenetic modifications, especially DNA methylation, histone acetylation, and histone methylation. We performed an in-depth analysis of the regulation of various epigenetic modifications on mammals from zygote formation to cleavage stage and blastocyst stage, and reviewed the modifications of key sites and their potential molecular mechanisms. In addition, we discuss the effects of nutrition (protein, lipids, and one-carbon metabolism) on epigenetic modification in embryos and emphasize the importance of various nutrients in embryonic development and epigenetics during pregnancy. Failures in epigenetic regulation have been implicated in mammalian and human early embryo loss and disease. With the use of reproductive technologies, it is becoming even more important to establish developmentally competent embryos. Therefore, it is essential to evaluate the extent to which embryos are sensitive to these epigenetic modifications and nutrition status. Understanding the epigenetic regulation of early embryo development will help us make better use of reproductive technologies and nutrition regulation to improve reproductive health in mammals., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society for Nutrition.)

Chlorophyll (Chl) degradation is a natural phenomenon that occurs during ripening in many fleshy fruit species, and also during fruit storage. The plant hormone ethylene is a key factor in promoting Chl degradation during fruit storage, but the mechanisms involved in this induction are largely unknown. In this study, an apple (Malus domestica) BEL1-LIKE HOMEODOMAIN transcription factor 7 (MdBEL7), potentially functioning as a transcriptional repressor of the Chl catabolic genes (CCGs), including MdCLH, MdPPH2 and MdRCCR2, was identified as a partner of the ethylene-activated U-box type E3 ubiquitin ligase MdPUB24 in a yeast library screen. Yeast-two-hybrid, co-immunoprecipitation and luciferase complementation imaging assays were then used to verify the interaction between MdBEL7 and MdPUB24. In vitro and in vivo ubiquitination experiments revealed that MdPUB24 functions as an E3 ubiquitin ligase to ubiquitinate MdBEL7, thereby causing its degradation through the 26S proteasome pathway. Transient overexpression of MdPUB24 in apple fruit led to a decrease in MdBEL7 abundance and increased expression of CCG genes, including MdCLH, MdPPH2 and MdRCCR2, as well as greater Chl degradation. Taken together, the data indicated that an ethylene-activated U-box type E3 ubiquitin ligase MdPUB24 directly interacts with and ubiquitinates MdBEL7. Consequent degradation of MdBEL7 results in enhanced expression of MdCLH, MdPPH2 and MdRCCR2, and thus Chl degradation during apple fruit storage. Our results reveal that an ethylene-MdPUB24-MdBEL7 module regulates Chl degradation by post-translational modification during apple fruit storage., (© 2021 Society for Experimental Biology and John Wiley & Sons Ltd.)

A rapid and simple method for the determination of 6 biogenic amines (BAs) in food was established on HPLC-MS /MS without derivatization. Samples were extracted with 5% perchloric acid and cleaned with n-hexane for lipid removal. The analytes were separated on Waters XBridge® HILIC (150 mm × 2.1 mm, 3.5 µm) and analyzed with multiple-reaction monitoring (MRM) mode after positive electrospray ionization on HPLC-MS/MS. Good linearity with high correlation coefficient was obtained between 10-1000 µg/L for cadaverine (CAD), putrescine (PUT), tyramine (TYR) and 2-phenylethylamine (2-PHE) and between 1-100 µg/L for histamine (HIS) and tryptamine (TRY), with the detection limits of the method ranging from 0.1 mg/kg for HIS and TRY, and 1.0 mg/kg for CAD, PUT, TYR and 2-PHE, which are under the residue limit of Chinese regulation. Spiking experiments demonstrated good recoveries between 70.2-114.6%, with relative standard deviations (RSDs) between 0.44-13.01%. This method was validated for BAs determination in liquor, fermented meat products, vegetable products, soybean products, dairy products, seafood and its derived products. These results promise high feasibility for BAs monitoring in various food with easy-to-operate and fast sample preparation process, stable analysis on HPLC-MS/MS without derivatization., 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 © 2021. Published by Elsevier B.V.)

Methods and Results: Herein, a comprehensive proteomic analysis was conducted on proliferative endometria from sows with low and normal reproductive performance (LRP and NRP, respectively). Enrichment analysis of differentially expressed proteins revealed alterations in endometrial remodeling, substance metabolism (mainly lipid, nitrogen, and retinol metabolism), immunological modulation, and insulin signaling in LRP sows. Importantly, aberrant lipid metabolite accumulation and dysregulation of insulin signaling were coincidently confirmed in endometria of LPR sows, proving an impaired insulin sensitivity. Furthermore, established high-fat diet- (HFD-) induced insulin-resistant mouse models revealed that uterine insulin resistance beginning before pregnancy deteriorated uterine receptivity and decreased implantation sites and fetal numbers. Mitochondrial biogenesis and fusion were decreased, and reactive oxygen species was overproduced in uteri from the HFD group during the implantation period. Ishikawa and JAR cells directly demonstrated that oxidative stress compromised implantation in vitro ., Conclusions: This study demonstrated that uterine insulin sensitivity impairment beginning before pregnancy resulted in implantation and fetal loss associated with oxidative stress induced by mitochondrial dysfunction., Competing Interests: The authors declare no competing interests., (Copyright © 2021 Meixia Chen et al.)

Maintaining ovarian steroidogenesis is of critical importance, considering that steroid hormones are required for successful establishment and maintenance of pregnancy and proper development of embryos and fetuses. Investigating the mechanism that butyrate modulates the ovarian steroidogenesis is beneficial for understanding the impact of lipid nutrition on steroidogenesis. Herein, we identified that butyrate improved estradiol and progesterone synthesis in rat primary ovarian granulosa cells and human granulosa KGN cells and discovered the related mechanism. Our data indicated that butyrate was sensed by GPR41 and GPR43 in ovarian granulosa cells. Butyrate primarily upregulated the acetylation of histone H3K9 (H3K9ac). Chromatin immune-precipitation and sequencing (ChIP-seq) data of H3K9ac revealed the influenced pathways involving in the mitochondrial function (including cellular metabolism and steroidogenesis) and cellular antioxidant capacity. Additionally, increasing H3K9ac by butyrate further stimulated the PPARγ/CD36/StAR pathways to increase ovarian steroidogenesis and activated PGC1α to enhance mitochondrial dynamics and alleviate oxidative damage. The improvement in antioxidant capacity and mitochondrial dynamics by butyrate enhanced ovarian steroidogenesis. Collectively, butyrate triggers histone H3K9ac to activate steroidogenesis through PPARγ and PGC1α pathways in ovarian granulosa cells., (© 2021 Federation of American Societies for Experimental Biology.)

One-carbon metabolism is involved in varieties of physiological processes in mammals, including nucleic acid synthesis, amino acid homeostasis, epigenetic regulation, redox balance and neurodevelopment. The current evidence linking levels of one-carbon nutrients during pregnancy to the development of oocytes, embryos, and placentas, as well as maternal and offspring health, is reviewed. The sources of mammalian one-carbon units, the pathways active in mammalian one-carbon metabolism, the maternal and fetal needs for one-carbon units and their functions during pregnancy are described. The demand for one-carbon metabolism is highest during pregnancy compared to the entire lifetime of a mammal. The primary types of one-carbon metabolism in mammals are the folate cycle, methionine cycle and transsulfuration pathway, which varies at different pregnancy stages (e.g., methylation programming of embryo, neural development of fetus, fetal growth and placenta development). Therefore, an overall consideration of one-carbon metabolism requirements for different pregnancy stages, is called for, specifically, the balance of all nutrients involved, not just one single nutrient in one-carbon metabolism. Moreover, the establishment of an ideal one-carbon metabolism requirement model is suggested according to the requirements for different pregnancy stages to support optimal pregnancy outcomes and maternal and offspring health., (© 2020 Wiley-VCH GmbH.)

Objectives: Early pregnancy loss is a major clinical concern in animal and human reproduction, which is largely influenced by embryo implantation. The importance of methionine for embryo implantation is widely neglected., Materials and Methods: We performed a series of experiments with primiparous rats fed diets containing different levels of methionine during early pregnancy to investigate the role of methionine in embryonic implantation and pregnancy outcomes, and used them to perform in vivo metabolic assessments and in vitro uterine explant culture. In addition, through transcriptome analysis and silencing the expression of cystathionine β-synthase (CBS, the key enzyme in transsulfuration pathway) and cell adhesion assay, we measured signalling within Ishikawa, pTr and JAR cells., Results: We determined the relevance and underlying mechanism of methionine on embryo implantation. We showed that methionine deprivation sharply decreased embryo implantation sites, expression of CBS and transsulfuration pathway end products, which were reversed by maternal methionine supplementation during early pregnancy. Moreover, we found CBS improved methionine-mediated cell proliferation and DNA synthesis by CBS inhibition or interference. In addition, transcriptome analysis also revealed that CBS influenced the signalling pathway-associated cell proliferation and DNA synthesis, as well as a correlation between CBS and methionine adenosyltransferase 2A (MAT2A), implying that MAT2A was possibly involved in cell proliferation and DNA synthesis. Further analysis revealed that MAT2A influenced S-adenosylmethionine receptor SAMTOR expression, and SAMTOR activated mTORC1 and its downstream S6K1 and CAD, ultimately enhancing DNA synthesis in the embryo and uterus., Conclusions: Taken together, these studies demonstrate that CBS and MAT2A improve methionine-mediated DNA synthesis through SAMTOR/mTORC1/S6K1/CAD pathway during embryo implantation., (© 2020 The Authors. Cell Proliferation Published by John Wiley & Sons Ltd.)

Microplastics (MPs) have become global environmental concern. However, the effects of environmental concentrations of MPs, singly or in combination with organic pollutants, on the early development of marine fish remain unclear. In this study, fertilized eggs of marine medaka (Oryzias melastigma) were exposed to polystyrene MPs (0, 2, 20, 200 μg/L) and/or phenanthrene (Phe, 50 μg/L) for 28 days. The results revealed that MPs were accumulated on the chorion and ingested by larvae from 2 days post-hatching. High levels of MPs (20 and 200 μg/L) decreased the hatchability, delayed the hatching time, and suppressed the growth, whereas Phe inhibited hatching and caused malformations in larvae. The presence of MPs at 20 and 200 μg/L did not alter the toxicity of Phe. By contrast, combined exposure to 2 μg/L MPs and Phe increased the hatchability by 25.8%, decreased malformation and mortality rates, and restored Phe-induced abnormal expressions of cardiac development-related genes. The reduced early developmental toxicity could be attributed to the decreased bioavailability and bioaccumulation of Phe by the low level of MPs. These findings contradicted the view that MPs would aggravate the toxicity of organic pollutants, and future studies are warranted to elucidate the ecological risks of marine MPs., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The superoxide anion (O2˙-) plays a crucial role in several physiological processes and many human diseases. Developing new methods for O2˙- detection in biological systems is very important. A FRET-based two-photon (TP) fluorescent probe with a ratiometric signal, TFR-O, was developed. A naphthalene derivative based TP fluorescent group was selected as the energy donor group, and a rhodol fluorescent group was chosen as the energy acceptor; the trifluoromethanesulfonate group was chosen as the recognition moiety. After reacting with O2˙-, the recognition moiety was removed and the fluorophore was released, leading to a fluorescence intensity decrease at the wavelength of 425 nm and a significant enhancement of the fluorescence intensity at 550 nm. The fluorescence intensity ratio between 550 and 425 nm (I550/I425) varied from 0.15 to 6.72, with the O2˙- concentration increasing from 0 to 50 μM. The detection limit of the TFR-O was 83 nM. Moreover, TFR-O was applied for detecting and imaging O2˙- in cells and liver tissues.

In the present study, pure zinc stents were implanted into the abdominal aorta of rabbits for 12 months. Multiscale analysis including micro-CT, scanning electron microscopy (SEM), scanning transmission electron microscopy (STEM) and histological stainings was performed to reveal the fundamental degradation mechanism of the pure zinc stent and its biocompatibility. The pure zinc stent was able to maintain mechanical integrity for 6 months and degraded 41.75 ± 29.72% of stent volume after 12 months implantation. No severe inflammation, platelet aggregation, thrombosis formation or obvious intimal hyperplasia was observed at all time points after implantation. The degradation of the zinc stent played a beneficial role in the artery remodeling and healing process. The evolution of the degradation mechanism of pure zinc stents with time was revealed as follows: Before endothelialization, dynamic blood flow dominated the degradation of pure zinc stent, creating a uniform corrosion mode; After endothelialization, the degradation of pure zinc stent depended on the diffusion of water molecules, hydrophilic solutes and ions which led to localized corrosion. Zinc phosphate generated in blood flow transformed into zinc oxide and small amounts of calcium phosphate during the conversion of degradation microenvironment. The favorable physiological degradation behavior makes zinc a promising candidate for future stent applications., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

BACKGROUND To study the effect of estrogen-related receptor α (ERRα) and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) on mesenchymal stem cells (MSCs) apoptosis, and further investigated its detailed molecular mechanisms in the absence of serum, hypoxia, and high glucose conditions. MATERIAL AND METHODS In our study, we first evaluated the expression rates of CD14, CD34, CD45, CD44, CD29, and Sca-1 surface markers on MSCs by flow cytometry. Then, the ability of osteogenic and fatty differentiation of MSCs was determined by osteogenic differentiation and adipogenesis reagent kit. Next, Annexin V-APC/7-AAD apoptosis kit was used for detecting the apoptosis rate of MSCs. RT-PCR and Western blotting were used for detection of mRNA expression and proteins expression, respectively. RESULTS Our data showed that the MSCs used in our study were capable of self-renewal and differentiating into many cell lineages, such as osteogenic differentiation and adipogenesis. Our results further showed that over-expression of PGC-1α could protect MSCs from apoptosis induced by rotenone. We also found that PGC-1α over-expression could enhance the expression of anti-apoptotic gene Bcl-2, and inhibit the expression of pro-apoptotic gene Bax in MSCs. In addition, our data demonstrated that PGC-1α could induce upregulation of Bcl-2 and further promote the survival of MSCs by interacting with ERRα. CONCLUSIONS In the absence of serum, hypoxia and high glucose conditions, PGC-1α can regulate the expression of Bcl-2 and promote the survival of MSCs via PGC-1α/ERRα interaction.

Tris(2,3-dibromopropyl) isocyanurate (TBC) is a novel brominated flame retardant (BFR) that is widely used to substitute the prohibited BFRs throughout the world. With the development of research, the potential environmental and ecological harms of TBC have been revealed. For sensitive and selective detecting TBC, an indirect competitive biotin-streptavidin-amplified enzyme-linked immunosorbent assay (BA-ELISA) has been established in this study. The small molecular TBC-hapten was synthesized first; it mimicked the chemical structure of TBC and possessed a secondary amine group. The as-obtained hapten was then conjugated with carrier proteins to prepare artificial antigen. After immunization, the anti-TBC polyclonal antibody was obtained from separating rabbit serum. The procedures of this BA-ELISA were optimized. Under the optimal conditions, the limit of detection (IC10) was 0.0067 ng/ml and the median inhibitory concentration (IC50) was 0.66 ng/ml. Cross-reactivity values of the BA-ELISA with the tested TBC analogues were ⩽5%. This immunoassay was successfully applied to determine the TBC residue in river water samples that were collected near a BFR manufacturing plant. Satisfactory recoveries (92.1-109.2%) were obtained. The results indicated that this proposed BA-ELISA is suitable for the rapid and sensitive determining of TBC in environmental monitoring., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Tetrabromobisphenol A (TBBPA) is a widely used brominated flame retardant. A sensitive and selective indirect competitive biotin-streptavidin-amplified enzyme-linked immunosorbent assay (BA-ELISA) was developed for detecting TBBPA. The optimal hapten of TBBPA was 2-(2,6-dibromo-4-(2-(3,5-dibromo-4-hydroxyphenly)propan-2-yl)) acetic acid. Several physiochemical factors that influence assay performance, such as optimal coupling concentration of immunogen and antibody, organic solvent, ionic strength, and pH, were studied and optimized. The limit of detection (IC10) was 0.027 ng/mL and the median inhibitory concentration (IC50) was 0.58 ng/mL. The BA-ELISA was highly selective, with low cross-reactivity with TBBPA analogs. Finally, the assay was used to detect TBBPA in electronic waste samples. The results are consistent with those using liquid chromatography, which proves that the proposed immunoassay is accurate and receptive. This BA-ELISA method is suitable for the rapid and sensitive screening of TBBPA in environmental monitoring., (© 2013 Published by Elsevier B.V.)