Effects of quercetin- and Lactiplantibacillus plantarum-containing bioactive films on physicochemical properties and microbial safety of grass carp.

In this study, the electrospinning technique was used to co-encapsulate Quercetin (Qu) and Lactiplantibacillus plantarum 1-24-LJ in PVA-based nanofibers, and the effect of bioactive films on fish preservation was evaluated at the first time. The findings indicated that both Lpb. plantarum 1-24-LJ and Qu were successfully in the fibers, and co-loaded fibers considerably outperformed single-loaded fiber in terms of bacterial survival and antioxidant activity. Following fish preservation using the loaded fibers, significant reductions were observed in TVB-N, TBARS, and microbial complexity compared to the control group. Additionally, the co-loaded fibers more effectively reduced the counts of H2S-producing bacteria and Pseudomonas. In the future, fibers with both active substances and LAB hold promise as a novel approach for fish preservation.

Electroacupuncture alleviates the symptom of depression in mice by regulating the cGAS-STING-NLRP3 signaling.

PURPOSE: To investigate the therapeutic effect of electroacupuncture (EA) on chronic and unpredictable mild stress (CUMS)-induced depression in mice and the underlying mechanism. METHODS: Male C57BL/6 mice were randomly divided into 6 groups: Control, CUMS, CUMS+EA-placebo, CUMS+EA, CUMS+ ad-NC, CUMS+ ad-cGAS-shRNA. CUMS was utilized to establish the depression model in mice. The behavioral changes were determined by the forced swimming, open field, and sucrose preference experiments. The pathological changes in the hippocampus tissue were evaluated by HE staining. The release of TNF-α, IL-1ß, IL-6, 5-HT, and NE in the hippocampus tissue was determined by ELISA. IBA-1 expression detected by the immunofluorescence was used to represent the activity of microglia. Western blot and RT-PCR were utilized to measure the expression of Bax, bcl-2, cGAS, STING, TBK1, IRF3, and NLRP3. RESULTS: The depression behavior in CUMS mice was significantly alleviated by the treatment of EA and cGAS-shRNA, accompanied by ameliorated hippocampus pathological changes, declined production of TNF-α, IL-1ß, and IL-6, elevated secretion of 5-HT and NE, and inhibition on the activity of microglia. Furthermore, significantly elevated expression level of Bax, cGAS, STING, TBK1, IRF3, and NLRP3 and declined expression level of bcl-2 were observed in the CUMS+EA and CUMS+ ad-cGAS-shRNA groups. CONCLUSIONS: EA significantly mitigated the symptom of depression in mice, which was closely associated with the repressed neuroinflammation, increased monoamine concentration, inactivated microglia, and inhibited cGAS-STING-NLRP3 signaling.

Enhanced Autophagy in Damaged Laminar Tissue of Acute Laminitis Induced by Oligofructose Overloading in Dairy Cows.

This study was aimed at determining the autophagy activity in the laminar tissue of dairy cows with oligofructose-induced laminitis. Twelve healthy non-pregnant Holstein cows were randomly divided into two groups of six cows each, entitled the control group and the oligofructose overload group (OF group), respectively. At 0 h, cows in the OF group were gavaged with oligofructose (17 g/kg BW) dissolved in warm deionized water (20 mL/kg BW) through an oral rumen tube, and the dairy cows in the control group were gavaged with the same volume of deionized water by the same method. At -72 h before, as well as 0 h, 6 h, 12 h, 18 h, 24 h, 36 h, 48 h, 60 h, and 72 h after perfusion, clinical evaluations of both groups were monitored. After 72 h, the laminar tissues of the dairy cows in both groups were collected to examine the genes and proteins. The gene expression of ATG5, ATG12, and Beclin1 significantly increased (p < 0.05), whereas that of P62 and mTOR significantly decreased (p < 0.01) in the OF group relative to the control group. The protein expression of Beclin-1 significantly increased (p < 0.05), while that of LC3II significantly decreased (p < 0.05) in the OF group relative to the control group. However, the protein expression of P62 non-significantly reduced (p > 0.05) in the OF group comparative to the control group. Furthermore, the distribution of the Beclin1 protein in the laminar tissue significantly increased (p < 0.01), while that of the P62 protein significantly decreased (p < 0.05) in the OF group than the control group. These findings indicate that the imbalanced gene and protein-level status of autophagy-related markers may be the basic cause for the failure of the epidermal attachment. However, a more detailed gene and protein-level study is needed to further clarify the role of autophagy in the pathogenesis of bovine laminitis.

Dual-Nanopipettes for the Detection of Single Nanoparticles and Small Molecules.

Nanopore sensing is blooming due to its label-free and high sensitivity features. As a novel nanopore, a droplet is formed at the orifice of a dual-nanopipette, which allows for the translocation of analytes through the two channels at a relatively low speed and the promotion of signal-to-noise ratio. However, nanopore sensing based on the principle of current blockage requires the pore size to be comparable to that of the single entity, which poses a huge challenge for the direct detection of small molecules. In this work, gold nanoparticles (Au NPs) modified with sulfhydryl poly(ethylene glycol) (PEG-SH) or aptamers were detected successfully. The size difference of Au NPs and the interaction between Au NPs and dual-nanopipettes could be distinguished sensitively. Furthermore, Au NPs modified with designed aptamers will produce different blocking current after capturing the corresponding small molecules (e.g., dopamine and serotonin). Even non-electroactive ions, such as potassium ions, can also be detected, which is difficult to sense based on redox reactions, and further illustrates that the change of surface properties of nanoparticles is responsible for the detection. This work expands the application of nanopipette sensing for Au NPs and provides a universal platform for the small-molecule detection, which has the potential application in biosensing.

Identifying Metal-Oxo/Peroxo Intermediates in Catalytic Water Oxidation by In Situ Electrochemical Mass Spectrometry.

Molecular catalysis of water oxidation has been intensively investigated, but its mechanism is still not yet fully understood. This study aims at capturing and identifying key short-lived intermediates directly during the water oxidation catalyzed by a cobalt-tetraamido macrocyclic ligand complex using a newly developed an in situ electrochemical mass spectrometry (EC-MS) method. Two key ligand-centered-oxidation intermediates, [(L2-)CoIIIOH] and [(L2-)CoIIIOOH], were directly observed for the first time, and further confirmed by 18O-labeling and collision-induced dissociation studies. These experimental results further confirmed the rationality of the water nucleophilic attack mechanism for the single-site water oxidation catalysis. This work also demonstrated that such an in situ EC-MS method is a promising analytical tool for redox catalytic processes, not only limited to water oxidation.

Antibacterial aerogels with nano­silver reduced in situ by carboxymethyl cellulose for fresh meat preservation.

Bacterial cellulose (BC) was used as a reinforcing agent, citric acid (CA) as a cross-linking agent, and CMC@AgNPs as antibacterial nanomaterials, in which CMC@AgNPs were reduced from AgNO3 in situ by carboxymethyl cellulose (CMC) as a reducing agent and stabilizer to fight microbial corruption. Its potential application in packaging fresh meat has been investigated. Results showed that the antibacterial CMC@AgNPs/BC/CA aerogels with excellent structural integrity and outstanding water absorption were developed by adding 0.3% BC and 0.25% CA. The CMC@AgNPs/BC/CA aerogel significantly reduced the color change and the total viable bacterial counts (TVC) in fresh meat after 7 days of refrigerated storage. The results indicated that CMC@AgNPs/BC/CA aerogels can effectively extend the shelf life of fresh meat, and can be used for meat packaging as a biologically active absorption pad.

Investigation of Dendrimer Transfer Behaviors at the Micro-Water/1,2-Dichloroethane Interface Facilitated by Dibenzo-18-Crown-6.

Trans-interfacial behaviors of multiple ionic species at the interface between two immiscible electrolyte solutions (ITIES) are of importance to biomembrane mimicking, chemical and biosensing, and interfacial molecular catalysis. Utilizing host-guest interaction to facilitate ion transfer is an effective and commonly used method to decrease the Gibbs energy of transfer of a target molecule. Herein, we investigated a facilitated ion transfer (FIT) process of poly(amidoamine)dendrimer (PAMAM, G0-G2) by dibenzo-18-crown-6 (DB18C6) at the microinterfaces between water and 1,2-dichloroethane (µ-W/DCE). Because of the host-guest interaction between a dendrimer and a ligand, negative shifts of the transfer potentials were observed using cyclic voltammetry or Osteryoung square wave voltammetry. From the FIT behavior of the dendrimer, we revealed that each DB18C6 could selectively coordinate with one amino group. We first evaluated the protonated status of the intermediate state (1:2) exactly under the conditions the dendrimer (G1) transfers across the interface using the electrochemical mass spectrometry (EC-MS)-hyphenated technique, which is much smaller than the protonated status in the water phase (1:8 to 14). Using the same methodology, we also studied the facilitated transfer behaviors of G0 and G2. Based on these results, we put forward the mechanism of the FIT process, which might involve a deprotonating process at the interface for higher-generation dendrimers.

Laminar Inflammation Responses in the Oligofructose Overload Induced Model of Bovine Laminitis.

Bovine laminitis causes substantial economic losses and animal welfare problems in dairy farms worldwide. Previously published studies have reported that the inflammatory response plays a central role in the pathogenesis of the disease. To our knowledge, inflammation associated with bovine laminitis induced by high levels of exposure to oligofructose (OF) has not been reported and characterized. In fact, the disease manifestations in this model closely approximate those of clinical laminitis. The objective of this study was to characterize the inflammatory response in OF-induced bovine laminitis. A total of 12 Chinese Holstein dairy heifers were utilized in this study. The heifers were randomly divided into two groups, treatment (n = 6) and control (n = 6). The treatment group heifers were administered OF solutions via a stomach tube (dose: 17 g/kg of body weight). Upon development of a lameness score of 2 with consecutive positive reactions in the same claw, they would be humanely euthanized. Control heifers were administered deionized water (dose: 2 L/100 kg of body weight) and humanely euthanized at 72 h. Real-time quantitative PCR (qPCR) assays were performed to determine the messenger RNA (mRNA) concentrations of inflammatory mediators in the lamellae. Concentrations of interleukin (IL)-1ß, IL-6, IL-8, C-X-C motif chemokine ligand-1 (CXCL-1), macrophage cationic peptide-2 (MCP-2), E-selectin, intercellular adhesion molecule-1 (ICAM-1), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase-1 (iNOS-1), and plasminogen activator inhibitor-1 (PAI-1) were significantly increased (P < 0.05) in the treatment group. No significant difference was found for tumor necrosis factor alpha (TNF-α), IL-10, CXCL-6, and MCP-1. These results demonstrated and characterized the laminar inflammatory response leading to the pathogenesis of bovine laminitis at the early stages.

Dynamic ROS Production and Gene Expression of Heifers Blood Neutrophil in a Oligofructose Overload Model.

Alimentary oligofructose (OF) overload can induce several diseases in cattle, such as ruminal acidosis, laminitis, and synovitis. The role of blood polymorphonuclear neutrophil (PMN) remains unclear during OF overload. The aim of this study was to investigate the dynamic changes in reactive oxygen species (ROS) production and the expression profile of genes in blood PMN in a model of OF overload. Twelve clinically healthy and non-pregnant Chinese Holstein heifers, aged between 18 and 26 mo, weighing 335-403 kg, BCS (5-point scale) ranges 2.7-3.3 were used for the experiments. OF heifers (n = 6) received 17 g/kg of BW oligofructose dissolved in 2 L/100 kg of BW tap water and the CON heifers (n = 6) received 2 L/100 kg of BW tap water. Blood PMN was isolated for each heifer 0, 6, 12, 18, 24, 36, 48, 60, and 72 h after administration. PMN was analyzed either by endogenous and phorbol myristate acetate (PMA)-induced ROS production or by quantitative real-time PCR. After 12 h, PMA-induced ROS production decreased, which was sustained until 48 h. The expressions of inflammation markers (IL1α, IL1ß, IL6, IL10, TNFα, STAT3, TLR4, MMP9, and HP) and eicosanoids (ALOX5, ALOX5AP, and PLA2G4A) were upregulated. The expression of adhesion and migration (CXCR2, CXCL8, CD62L, ITGA4, ITGAM, and ITGB2) in OF heifers was increased compared with CON heifers. The expression of oxidative stress (SOD2 and S100A8) was upregulated, while SOD1 and MPO were downregulated. In metabolism and receptor genes, the expressions of GRα and INSR decreased after 12 h, while Fas increased until 6 h and then decreased at 18 h. The expression of LDHA and PANX1 did not show any differences after OF overload. These findings indicate that OF overload induced systemic activation of PMN, which provides a step toward a better understanding of the role of innate immune responses in response to oral OF administration.

The changes of inflammatory mediators and vasoactive substances in dairy cows' plasma with pasture-associated laminitis.

BACKGROUND: Hoof disease is one of the three major diseases that often occur in dairy cows. The impact of this disease on dairy farming is second only to mastitis. Laminitis is a diffuse, aseptic, serous, non-purulent inflammation of the dermal papillae and vascular layers of the cow's hoof wall. In the pasture, laminitis occurs mostly in the laminae, that is, inside the hoof shell. No lesions can be seen on the surface. Therefore, laminitis cannot attract the attention of veterinarians. However, laminitis has become a major factor that seriously affects the health and welfare of dairy cows, making it an important cause of hindering the performance of dairy cows. METHODS: The study was conducted at a dairy farm in Harbin, Heilongjiang province, China. We selected a sample of the laminitis cows based on the veterinary diagnosis, took blood from the jugular vein and then separated the plasma, and measured the index with the Elisa kit. In this study, the markers of inflammatory and vasoactive substances status in dairy cows consisted of subclinical laminitis (SCL, n = 20), chronic laminitis (CL, n = 20) and healthy dairy cows (CON, n = 20) under the local management conditions were investigated. RESULTS: Compared with healthy cattle, HIS, IL-6, LPS, and TNF-α in subclinical laminitis group significantly increased (P < 0.05), especially HIS, LPS, TNF-α (P < 0.01); in chronic laminitis cows, COX-2, HIS, IL-6, LPS, and TNF-α increased significantly (P < 0.05), especially COX-2, HIS, TNF-α (P < 0.01). iNOS (P < 0.05), TXB2 (P < 0.01) in chronic laminitis cows had significantly increased. CONCLUSION: This study reported for the first time that pasture laminitis was divided into subclinical laminitis and clinical chronic laminitis. Through research on the inflammatory factors and vasoactive substances of dairy cows, it is found that there is a close relationship between them, which affects the metabolic cycle of dairy cows. These indicators are abnormally expressed and cause hoof microcirculation disorders.

Comparative analysis of cutaneous bacterial communities of farmed Rana dybowskii after gentamycin bath.

INTRODUCTION: Pathogenic bacteria limit the success of Rana dybowskii breeding. Gentamicin is used to treat R. dybowskii disease. To understand the effects of gentamicin on the composition and structure of the cutaneous bacterial community of R. dybowskii, three groups (control, gentamicin and recovery) were established in this study. MATERIALS & METHODS: The V3-V4 hypervariable region of the 16S rRNA gene was analyzed in samples by high-throughput sequencing. Alpha diversity and beta diversity were evaluated to compare the cutaneous bacterial community diversity. RESULTS: A total of 1,159,668 valid sequences and 3,132 operational taxonomic units (OTUs) were obtained from these three experimental groups. The number of OTUs obtained in the control group, gentamicin group and recovery group were 2,194, 2,288, and 2,047, respectively, and the number of shared OTUs was 1,313. The alpha diversity of the cutaneous bacterial community was not significantly affected by gentamicin, while beta diversity was significantly affected. DISCUSSION & CONCLUSIONS: The effect of a gentamicin bath on relative species abundance was greater than the effect on the species composition. The changes in Proteobacteria, Acinetobacter, and Chryseobacterium were significant, and reductions were observed after the recovery period. Six potentially pathogenic genera were detected, including Aeromonas, Citrobacter, Chryseobacterium, Pseudomonas, Staphylococcus, and Streptococcus. Among them, Aeromonas and Chryseobacterium were significantly inhibited by the gentamicin bath. The results of this study provide a theoretical basis for the application of gentamicin in R. dybowskii breeding.

Anodic oxidation triggered divergent 1,2- and 1,4-group transfer reactions of ß-hydroxycarboxylic acids enabled by electrochemical regulation.

We report a set of electrochemically regulated protocols for the divergent synthesis of ketones and ß-keto esters from the same ß-hydroxycarboxylic acid starting materials. Enabled by electrochemical control, the anodic oxidation of carboxylic acids proceeded in either a one-electron or a two-electron pathway, leading to a 1,4-aryl transfer or a semipinacol-type 1,2-group transfer product with excellent chemoselectivity. The 1,4-aryl transfer represents an unprecedented example of carbon-to-oxygen group transfer proceeding via a radical mechanism. In contrast to previously reported radical group transfer reactions, this 1,4-group transfer process features the migration of electron-rich aryl substituents. Furthermore, with these chemoselective electrochemical oxidation protocols, a range of ketones and ß-keto esters including those possessing a challenging-to-access medium-sized ring could be synthesized in excellent yields.

Fabrication of Tris(bipyridine)ruthenium(II)-Functionalized Metal-Organic Framework Thin Films by Electrochemically Assisted Self-Assembly Technique for Electrochemiluminescent Immunoassay.

A simple strategy for one-step fabrication of tris(bipyridine)ruthenium(II) (Ru(bpy)32+)-functionalized metal-organic framework (Ru-MOF) thin films using a self-assembly approach assisted by an electrochemical way was introduced. In this protocol, the electrochemically driven cooperative reaction of Ru(bpy)32+ as an electrochemiluminescent (ECL) probe and a structure-directing agent, trimesic acid (H3btc) as a ligand, and Zn(NO3)2 as the Zn2+ source leads to an one-step and simultaneous synthesis and deposition of the MOF onto the electrode surface. Characterization of the Ru-MOF thin films was performed with scanning electron microscopy, Fourier transform infrared, and X-ray photoelectron spectroscopy. Scanning ion conductance microscopy was specially applied in situ to image the topography and thickness of the Ru-MOF thin films. The Ru-MOF thin films as a sensing platform show excellent ECL behavior because of plenty of Ru(bpy)32+ molecules encapsulated in the frameworks. On the basis of the Ru-MOF modified electrodes, an ultrasensitive label-free ECL immunosensing method for the human heart-type fatty-acid-binding protein has been developed with a wide linear response range (150 fg mL-1-150 ng mL-1) and a very low limit of detection (2.6 fg mL-1). The prepared immunosensor also displayed excellent stability and good specificity in the test of practical samples.

Nano-bacterial cellulose/soy protein isolate complex gel as fat substitutes in ice cream model.

The influences of nano-bacterial cellulose (Nano-BC)/soy protein isolate (SPI) complex gel on the textural, rheological, and sensory properties of the ice cream model were investigated. Nano-BC/SPI mixtures with different ratios (Nano-BC:SPI, 1:20, 1:15, 1:10, and 1:5 w/w) were prepared with constant total solid content (16%). Compared with pure SPI, the thermal stability, textural, rheological and emulsifying properties of nano-BC/SPI mixtures were improved. Microstructure of nano-BC/SPI complex gels showed that low doses of nano-BC (1:20, 1:15, and 1:10) had good compatibility with SPI matrix according to images of confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Nano-BC/SPI (1:20) had the most similar textural properties to the cream. When 20% of nano-BC/SPI (1:20) mixture was added into ice cream as the cream substitute, the anticipated ice cream with low calorie, melting resistance, and good textural properties was achieved.

Ionic Current Behaviors of Dual Nano- and Micropipettes.

Ionic current rectification (ICR) phenomena within dual glass pipettes are investigated for the first time. We demonstrate that the ionic flow presents different behaviors in dual nano- and micropipettes when the two channels are filled with the same electrolyte KCl and hung in air. Bare dual nanopipettes cannot rectify the ionic current because of their geometric symmetry, but the ICR can be directly observed based on bare dual micropipettes. The phenomena based on dual micropipettes could be explained by the simulation of the Poisson-Nernst-Plank equation. After modification with different approaches, the dual nanopipettes have asymmetric charge patterns and show various ICR behaviors. They have been successfully employed to fabricate various nanodevices, such as ionic diodes and bipolar junction transistors. Due to the simple and fast fabrication with high reproducibility, these dual pipettes can provide a novel platform for controlling ionic flow in nano- and microfluidics, fabrication of novel nanodevices, and detection of biomolecules.

Ionic current rectification in organic solutions with quartz nanopipettes.

The study of behaviors of ionic current rectification (ICR) in organic solutions with quartz nanopipettes is reported. ICR can be observed even in organic solutions using quartz pipettes with diameters varied from several to dozens of nanometers, and the direction of ICR is quite different from the ICR observed in aqueous phase. The influences of pore size, electrolyte concentration, and surface charge on the ICR have been investigated carefully. Water in organic solutions affects the direction and extent of ICR significantly. Mechanisms about the formation of an electrical double layer (EDL) on silica in organic solutions with different amount of water have been proposed. An improved method, which can be employed to detect trace water in organic solutions, has been implemented based on Au ultramicroelectrodes with cathodic differential pulse stripping voltammetry.