ANGPTL2 knockdown induces autophagy to relieve alveolar macrophage pyroptosis by reducing LILRB2-mediated inhibition of TREM2.

Acute lung injury (ALI) is featured with a robust inflammatory response. Angiopoietin-like protein 2 (ANGPTL2), a pro-inflammatory protein, is complicated with various disorders. However, the role of ANGPTL2 in ALI remains to be further explored. The mice and MH-S cells were administrated with lipopolysaccharide (LPS) to evoke the lung injury in vivo and in vitro. The role and mechanism of ANGPTL was investigated by haematoxylin-eosin, measurement of wet/dry ratio, cell count, terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labeling, reverse transcription quantitative polymerase chain reaction, immunofluorescence, enzyme-linked immunosorbent assay, detection of autophagic flux and western blot assays. The level of ANGPTL2 was upregulated in lung injury. Knockout of ANGPTL2 alleviated LPS-induced pathological symptoms, reduced pulmonary wet/dry weight ratio, the numbers of total cells and neutrophils in BALF, apoptosis rate and the release of pro-inflammatory mediators, and modulated polarization of alveolar macrophages in mice. Knockdown of ANGPTL2 downregulated the level of pyroptosis indicators, and elevated the level of autophagy in LPS-induced MH-S cells. Besides, downregulation of ANGPTL2 reversed the LPS-induced the expression of leukocyte immunoglobulin (Ig)-like receptor B2 (LILRB2) and triggering receptor expressed on myeloid cells 2 (TREM2), which was reversed by the overexpression of LILRB2. Importantly, knockdown of TREM2 reversed the levels of autophagy- and pyroptosis-involved proteins, and the contents of pro-inflammatory factors in LPS-induced MH-S cells transfected with si ANGPTL2, which was further inverted with the treatment of rapamycin. Therefore, ANGPTL2 silencing enhanced autophagy to alleviate alveolar macrophage pyroptosis via reducing LILRB2-mediated inhibition of TREM2.

Combined analysis of transcriptomics with metabolomics provides insights into the resistance mechanism in winter jujube using L-Methionine.

Black rots lead to great economic losses in winter jujube industry. The objective of this research was to delve into the underlying mechanisms of enhanced resistance of winter jujube fruit to black rot by L-Methionine (Met) treatment. The findings revealed that the application of Met significantly curtailed lesion diameter and decay incidence in winter jujube fruit. The peroxidase (POD) activity in the Met-treated jujubes was 3.06-fold that in the control jujubes after 4 d of treatment. By day 8, the activities of phenylalanine ammonia-lyase (PAL), chitinase (CHI) and ß-1,3-glucanase (GLU) in the Met-treated jujubes had surged to their zenith, being 1.39, 1.22, and 1.52 times in the control group, respectively. At the end of storage, the flavonoid and total phenol content remained 1.58 and 1.06 times than that of the control group. Based on metabolomics and transcriptomics analysis, Met treatment upregulated 6 key differentially expressed metabolites (DEMs) (succinic acid, trans-ferulic acid, salicylic acid, delphinium pigments, (S)-abscisic acid, and hesperidin-7-neohesperidin), 12 key differentially expressed genes (DEGs) (PAL, CYP73A, COMT, 4CL, CAD, POD, UGT72E, ANS, CHS, IAA, TCH4 and PR1), which were involved in phenylpropanoid biosynthesis pathway, flavonoid biosynthesis pathway and plant hormone signal transduction pathway. Further analysis revealed that the most of the enzymes, DEMs and DEGs in this study were associated with both antioxidant and disease resistance. Consequently, Met treatment enhanced disease resistance of winter jujube fruit by elevating antioxidant capacity and triggering defense response. This study might provide theoretical support for utilizing Met in the management and prevention of post-harvest black rot in winter jujube.

Inhibitory Effect of L-Methionine on Alternaria alternata Based on Metabolomics Analysis.

Alternaria alternata is the main pathogenic fungus of postharvest black spots in fruits and vegetables. This study aimed to explore the antifungal activity of methionine on A. alternata in vitro and to reveal related antifungal mechanisms through a metabolomics analysis. The results showed that the inhibitory effects of L-methionine (Met) treatment on mycelium growth, spore germination, and the germ tube elongation of A. alternata were enhanced with an increase in the Met concentration, but the inhibitory effects decreased when the Met concentration was higher than 50 mmolL-1. The results of propidium iodide staining and scanning electron microscopy showed that the Met treatment damaged the plasma membrane integrity of the A. alternata spores and caused an irreversible deformation of mycelium. In addition, after the Met treatment, the leakage of electrolytes, nucleic acid, and proteins in the A. alternata cells was significantly higher than that in the control group, indicating that the Met treatment increased the permeability of the cell membranes. Eighty-one different metabolites, divided into seven categories, were identified through the metabolomics analysis, including forty-three downregulated metabolites and thirty-eight upregulated metabolites. Among them, these differential metabolites were mainly involved in amino acid synthesis and metabolism, the pentose phosphate pathway, and the TCA cycle. Therefore, the antifungal effect of the Met treatment on A. alternata was mainly to damage the integrity of the cell membranes, make nucleic acid and protein contents leak, and affect the TCA cycle, carbohydrate metabolism, amino acid synthesis metabolism, and the metabolic pathways associated with cell membrane biosynthesis. Thus, the growth and development of A. alternata were inhibited. The research enriched the investigation of the effect of the antifungal mechanism of Met treatment on A. alternata and provided a theoretical basis for the application of Met to prevent and treat postharvest black spots in fruits and vegetables.

Characterizations of Water-Soluble Chitosan/Curdlan Edible Coatings and the Inhibitory Effect on Postharvest Pathogenic Fungi.

This study focused on developing a composite coating comprising water-soluble chitosan (CTS) and curdlan (CUR). Cherry tomatoes served as the test material for assessing the preservative efficacy of these coatings. The incorporation of CUR markedly enhanced the coating's surface properties, refined its molecular structure, and improved its tensile strength and elongation at break. Additionally, the coating demonstrated enhanced permeability to water vapor, oxygen, and carbon dioxide and improved light transmission. The storage experiment, conducted at 25 ± 1 °C with a relative humidity of approximately 92% over 10 days, revealed that the CTS/CUR composite coating at a 1:1 ratio significantly outperformed the individual CTS or CUR coating and uncoated samples in maintaining the quality of postharvest cherry tomatoes. The 1:1 CTS/CUR composite coating demonstrated superior preservative effects. This study suggested that water-soluble chitosan/curdlan composite coatings have considerable potential for use in the preservation of postharvest fruits and vegetables.

Characterization and Antioxidant and Antibacterial Activities of Carboxymethylated Tamarind Seed Polysaccharide Composite Films Incorporated with ε-Polylysine and Their Application in Fresh-Cut Green Bell Pepper Preservation.

Traditional petroleum-based food-packaging materials have poor permeability, limited active packaging properties, and difficulty in biodegradation, limiting their application. We developed a carboxymethylated tamarind seed polysaccharide composite film incorporated with ε-polylysine (CTPε) for better application in fresh-cut agricultural products. The CTPε films exhibit excellent water vapor barrier properties, but the mechanical properties are slightly reduced. Fourier transform infrared spectroscopy and X-ray diffraction spectra indicate the formation of hydrogen bonds between ε-PL and CTP, leading to their internal reorganization and dense network structure. With the increase of ε-PL concentration, composite films showed notable inhibition of postharvest pathogenic fungi and bacteria, a significant enhancement of 2,2'- azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical-scavenging activity, and gradual improvement of wettability performance. Cytotoxicity experiments confirmed the favorable biocompatibility when ε-PL was added at 0.3% (CTPε2). In fresh-cut bell pepper preservation experiments, the CTPε2 coating effectively delayed weight loss and malondialdehyde increase preserved the hardness, color, and nutrients of fresh-cut peppers and prolonged the shelf life of the fresh-cut peppers, as compared with the control group. Therefore, CTPε composite films are expected to be a valuable packaging material for extending the shelf life of freshly cut agricultural products.

Inhibition Mechanism of Water-Soluble Chitosan-Curdlan Composite Coating on the Postharvest Pathogens of Serratia marcescens and Pseudomonas syringae in Cherry Tomatoes.

Cherry tomatoes, a very popular fruit, are highly susceptible to microbial infestation, which cause significant economic losses. In order to preserve cherry tomatoes better, we treat them with a Chitosan (CTS) and Curdlan (CUR) composite coating. The lowest inhibitory concentration of CTS/CUR composite coating on Serratia marcescens and Pseudomonas syringae, the growth curves, and the changes of the cell lysis rate were determined to explore the inhibitory mechanism of CTS/CUR composite coating on Serratia marcescens and Pseudomonas syringae and the microscopic morphology of Serratia marcescens and Pseudomonas syringae was observed using scanning electron microscopy at the same time. The results showed that the CTS/CUR composite coating could effectively inhibit the growth of Serratia marcescens and Pseudomonas, and the inhibitory effect reflected the concentration-dependent characteristics. The electron microscopy results indicated that the inhibition of Serratia marcescens and Pseudomonas syringae by the CTS/CUR composite coating might originate from its disruptive effect on the cell wall and cell membrane of the bacterium.

Development and Characterization of Modified Gelatin-Based Cling Films with Antimicrobial and Antioxidant Activities and Their Application in the Preservation of Cherry Tomatoes.

The utilization of functional cling films presents a promising approach to alleviate post-harvest spoilage caused by microbial activity, oxidative metabolism, and moisture loss in agricultural products. To overcome the environmental problems of conventional packaging materials, in this study, we developed functional fruit and vegetable cling films based on glycidyltrimethylammonium chloride and rosemarinic acid cross-linked gelatin (RQ-GEL). The results indicate that the prepared RQ-GEL film possesses excellent UV light barrier properties and mechanical performance. RQ-GEL inhibited S. aureus and E. coli by 93.79% and 92.04%, respectively. DPPH and ABTS free radical scavenging activities were as high as 87.69% and 84.6%. In the cherry tomato preservation experiment, when compared to uncovered samples, the RQ-GEL group had a 29.77% reduction in weight loss and a significant 26.92% reduction in hardness. Meanwhile, the RQ-GEL group delays the decline of fruit total soluble solids and titratable acidity content, and prolongs the preservation period of cherry tomatoes. Hence, RQ-GEL cling film is poised to emerge as a promising packaging material for the post-harvest preservation of agricultural products.

Antifungal Activities of L-Methionine and L-Arginine Treatment In Vitro and In Vivo against Botrytis cinerea.

Gray mold caused by Botrytis cinerea is a common postharvest fungal disease in fruit and vegetables. The prevention and treatment of postharvest gray mold has been one of the hot research issues addressed by researchers. This study aimed to investigate the effect of L-methionine and L-arginine on Botrytis cinerea in vitro and on cherry tomato fruit. The results of the in vitro experiment showed that L-methionine and L-arginine had significant inhibitory effects on the mycelial growth and spore germination of Botrytis cinerea, and the inhibitory effects were enhanced with increasing L-methionine or L-arginine concentration. In addition, L-methionine and L-arginine treatment increased the leakage of Botrytis cinerea electrolytes, proteins and nucleic acids. The experiment involving propidium iodide staining and malondialdehyde content assay also confirmed that L-methionine and L-arginine treatment could lead to cell membrane rupture and lipid peroxidation. The results of scanning electron microscopy further verified that the morphology of hyphae was damaged, deformed, dented and wrinkled after treatment with L-methionine or L-arginine. Fruit inoculation experiments displayed that L-methionine and L-arginine treatments significantly inhibited the occurrence and development of gray mold in postharvest cherry tomato. Therefore, treatment with L-methionine or L-arginine might be an effective means to control postharvest gray mold in fruit and vegetables.

Preparation of Fresh-Keeping Paper Using Clove Essential Oil through Pickering Emulsion and Maintaining the Quality of Postharvest Cherry Tomatoes.

This study focused on developing a Pickering emulsion fresh-keeping paper that contained clove essential oil (CEO). Cherry tomatoes served as the test material for assessing the preservative efficacy of fresh-keeping paper. The results showed that Pickering emulsion had strong stability. Additionally, the fresh-keeping paper had a good antioxidant activity and sustained-release effect on CEO. In terms of the preservation effect, 0.75 wt% CEO Pickering emulsion paper reduced the decay incidence and weight loss of cherry tomatoes during 12-day storage. Fresh-keeping paper could also play a positive role in protecting the sensory index and color difference of tomatoes. It slowed the decline rate of soluble solid concentration (SSC) and titrable acid (TA). The vitamin C (Vc) and hardness of preserved tomatoes using fresh-keeping paper were maintained at a high level. The paper also inhibited the growth of microorganisms significantly. Therefore, 0.75 wt% CEO Pickering emulsion fresh-keeping paper displayed considerable potential for application in the preservation of postharvest fruits and vegetables. It is a novel fruit and vegetable preservation material worthy of development.

Black necrosis of the glans penis associated with calciphylaxis: A case report.

RATIONALE: Calciphylaxis, known as calcific uremic arteriolopathy, is a rare cause of dry gangrene. Despite an increase in the clinical recognition of demographic characteristics and risk factors associated with calciphylaxis, it remains a poorly understood disease with high mortality. PATIENT CONCERNS AND DIAGNOSES: We present a 45-year-old man, who was diagnosed with calciphylaxis disease, with a history of diabetes mellitus, end-stage renal disease and cirrhosis with a half-month evolution of painful dry gangrene on his glans penis and scrotum. The patient also presented with gangrene of fingers. INTERVENTIONS AND OUTCOMES: The patient and his family opted for palliative care. However, he died eventually. LESSONS: This case contributed to the current understanding of calciphylaxis. Since no standard treatment is available and the prognosis remained poor, early, and accurate diagnosis of calciphylaxis is important. We here report the current case and provide data for the diagnosis and treatment of this kind of disease.

Heme oxygenase-1/carbon monoxide signaling participates in the accumulation of triterpenoids of Ganoderma lucidum.

Ganoderic triterpenoids (GTs) are the primary bioactive constituents of the Basidiomycotina fungus, Ganoderma lucidum. These compounds exhibit antitumor, anti-hyperlipidemic, and immune-modulatory pharmacological activities. This study focused on GT accumulation in mycelia of G. lucidum mediated by the heme oxygenase-1 (HO-1)/carbon monoxide (CO) signaling. Compared with the control, hemin (10 µmol/L) induced an increase of 60.1% in GT content and 57.1% in HO-1 activity. Moreover, carbon monoxide-releasing molecule-2 (CORM-2), CO donor, increased GT content by 56.0% and HO-1 activity by 18.1%. Zn protoporphyrin IX (ZnPPIX), a specific HO-1 inhibitor, significantly reduced GT content by 26.0% and HO-1 activity by 15.8%, while hemin supplementation reversed these effects. Transcriptome sequencing showed that HO-1/CO could function directly as a regulator involved in promoting GT accumulation by regulating gene expression in the mevalonate pathway, and modulating the reactive oxygen species (ROS) and Ca2+ pathways. The results of this study may help enhance large-scale GT production and support further exploration of GT metabolic networks and relevant signaling cross-talk.

Ultrasonic and microwave treatment improved jujube juice yield.

The methods to leaching juice from dried jujube using six treatments and some factors related with juice yield such as total soluble solids (TSS), pectinase activity, pectin contents, galacturonic acid, and the microstructure morphology of cell were investigated. Six treatments including natural leaching, ultrasonic, microwave, ultrasonic before microwave, ultrasonic after microwave, and pressing directly were applied to extract juice. The group which treating with ultrasonic before microwave displayed its total soluble solids (TSS) was 16 °Brix, which was 6.67% higher than that of natural leaching. And its total soluble solids (TSS) reached to equilibrium in 2 hr, which was faster than that of natural leaching. The mechanism of improving the production efficiency of juice yield using ultrasonic combined with microwave was explored accordingly. The content of pectin and galacturonic acid increased by 58.52% and 59.01%, respectively, which were the highest among all samples. The activity of pectinase was 9.71 µg/(h·g), which was significantly decreased 40.23% as compared to natural leaching. And the treated cells became shriveled and pitted, which led to the leakage of the contents of cell. Thus, the result showed that treating with ultrasonic before microwave displayed the best juice yield. Ultrasonic cooperate with microwave was an efficient method to leaching juice from dried jujube.

A quencher-free 2-aminopurine modified hairpin aptasensor for ultrasensitive detection of Ochratoxin A.

A sensitive, efficient and quencher-free fluorescence aptasensor to detect Ochratoxin A (OTA) based on aptamer, 2-aminopurine (2AP) labeled Oligonucleotide sequence, as well as exonuclease I (Exo I) activity was developed. In which the aptamer specific to OTA was modified into a hairpin structure, and 8 bases at the 3' ends are exposed (H); also, 2AP is embedded in the oligonucleotide complementary to the 8 bases (2AP-probe).The detection principle based on 2AP-probe could be bonded to its complementary sequence and quenches the fluorescence of 2AP; The aptamer has a stronger affinity for the target than its complementary sequence; Exo I can dissociate single-stranded DNA and has little effect on double-stranded DNA as well as folded DNA. In the absence of OTA, the fluorescence of 2AP is quenched due to the complementary pairing of H and 2AP-probe; in the presence of OTA, H selective binding target is detached from 2AP-probe, and the fluorescence of 2AP is slightly restored. Moreover, when the Exo I is added to the detection system, 2AP-probe is dissociated by the Exo I to release the free 2AP, and the fluorescence of the system is further enhanced thereby realizing the detection of OTA. The detection limit of the aptasensor was low as 0.03 nM with a linear range of 0.5-100 nM. Moreover, the aptasensor has good selectivity and practicability and also has good potential in realizing the detection of toxic and harmful substances in food complex matrices.

Determination of patulin in apple juice using magnetic solid-phase extraction coupled with high-performance liquid chromatography.

An efficient magnetic sorbent consisting of benzofuran-2-carboxylic acid-loaded magnetic nanocomposite was successfully synthesised for pre-concentration of patulin from apple juice. The prepared magnetic nanocomposite was characterised by scanning electron microscopy, transmission electron microscopy and Fourier-transform infrared spectroscopy. Determination of enriched patulin was performed by high-performance liquid chromatography. The best adsorption conditions were 40 mg of sorbent, 50 ml of apple juice sample, pH 5, ambient temperature and 25 min; the elution conditions were 500 µl methanol, pH 5, ambient temperature, and 4 min. Under optimised conditions, pre-concentration factor was 100, linearity range was 1-400 µg l-1 of patulin, limit of detection was 0.15 µg l-1 and limit of quantification was 0.5 µg l-1. When samples were determined 20 times, the recovery was 93.9-102.6% and the relative standard deviation was below 5.3%. In terms of proposed procedure, the developed method was successfully applied for patulin detection in apple juice samples.

Determination of tetracyclines in bovine milk using laccaic acid-loaded magnetite nanocomposite for magnetic solid-phase extraction.

An efficient magnetic sorbent of laccaic acid-loaded magnetite nanocomposite was successfully synthesized for extraction and preconcentration of tetracyclines (TCs) from bovine milk sample. The synthesized nanocomposite was characterized by scanning electron microscopy and Fourier transform-infrared spectroscopy. Determination of the extracted TCs was performed by high performance liquid chromatography. Parameters affecting the efficiency of magnetic solid-phase extraction were studied and optimized. The best adsorption conditions were 80 mg of sorbent, 50 mL of sample volume, pH 7, 45°C and 20 min; the elution conditions were 0.5 mL of acetonitrile, pH 5.5, 75°C and 5 min. Under optimized conditions, the preconcentration factor of TCs was 100. The calibration curves were linear at a concentration range of 10-300 µg/kg for oxytetracycline and TC, and 5-300 µg/kg for chlortetracycline and doxycycline. The limit of detection was from 3.2 to 7.1 µg/kg, the limit of quantification was from 10.7 to 23.7 µg/kg, the relative standard deviation was between 2.8 and 5.3 and the recoveries were between 95.2% and 106.2%. In terms of the proposed procedure, the developed method was successfully applied for TCs detection in bovine milk samples.

Magnetic solid-phase extraction coupled with HPLC for the determination of Allura Red in food and beverage samples.

A magnetic solid-phase extraction (MSPE) protocol prior to HPLC was developed for the extraction and determination of Allura Red in food samples. Magnetic nanoparticles were coated with tetraethylorthosilicate and 3-aminopropyltriethoxysilane and modified by graphene. Scanning electron microscopy, transmission electron microscopy and Fourier-transform infrared spectrometry were used to characterise the graphene-functionalised sorbents; and the main parameters affecting the extraction such as sample volume, temperature, pH and time were investigated and established. Under optimised conditions, the pre-concentration factor of Allura Red was 200, and the calibration curve was linear at a concentration range of 5-1500 µg kg-1. The LOD was 2 µg kg-1, the limit of quantification was 7 µg kg-1, and the relative standard deviation was 3.3%. The prepared MSPE procedure was simple and fast, and it was successfully applied for the determination of Allura Red in beverages, candy and jelly.