Development and application of an indirect ELISA for the serological detection of bovine viral diarrhea virus infection based on the protein E2 antigen.

BACKGROUND: Bovine viral diarrhea virus (BVDV) causes continuous economic losses to the livestock industry. Monitoring antibodies with enzyme-linked immunosorbent assay (ELISA) is a valuable tool to ensure the purification of BVDV in cattle. However, currently available ELISA kits based on the whole BVDV virion are both costly and time-consuming. The E2 protein has good immunogenicity, induces the secretion of neutralizing antibodies and is an essential immunogen for serological detection. METHODS AND RESULTS: We developed a novel recombinant E2 protein-based indirect ELISA (rE2-iELISA) and conducted a serological survey for BVDV antibodies in 2021-2022 in Beijing, China. The results showed that E2 protein was successfully expressed with high immunogenicity and the optimal rE2-iELISA displayed high sensitivity, reproducibility and specificity. Clinical testing of 566 serum specimens indicated that 318 BVDV positive samples and 194 BVDV negative samples were tested by rE2-iELISA and the IDEXX BVDV ELISA-Ab kit, with a positive coincidence rate of 93.3%, a negative coincidence rate of 86.3%, and an overall coincidence rate of 90.5%. CONCLUSION: This study established an rE2-iELISA method, which is a highly sensitive, specific and robust ELISA-test validated to detect anti-BVDV antibodies. These findings indicate that the newly developed rE2-iELISA method has the potential to be used as a rapid, reliable and cost-effective screening tool for BVDV infection and provides technical support for the evaluation of vaccine efficacy in cattle herds in the future.

Supercharged CO2 Photothermal Catalytic Methanation: High Conversion, Rate, and Selectivity.

To overcome the thermodynamic and kinetic impediments of the Sabatier CO2 methanation reaction, the process must be operated under very high temperature and pressure conditions, to obtain an industrially viable conversion, rate, and selectivity. Herein, we report that these technologically relevant performance metrics have been achieved under much milder conditions using solar rather than thermal energy, where the methanation reaction is enabled by a novel nickel-boron nitride catalyst. In this regard, an in situ generated HOB⋅⋅⋅B surface frustrated Lewis's pair is considered responsible for the high Sabatier conversion 87.68 %, reaction rate 2.03 mol gNi -1 h-1 , and near 100 % selectivity, realized under ambient pressure conditions. This discovery bodes well for an opto-chemical engineering strategy aimed at the development and implementation of a sustainable 'Solar Sabatier' methanation process.

A Bovine Viral Diarrhea Virus Type 1c Strain in China: Isolation, Identification, and Assessment of Pathogenicity in Rabbits.

Bovine viral diarrhea virus (BVDV) is an important animal pathogen and has a negative economic impact on cattle industries worldwide. In this study, the BVDV strain named BJ175170 was detected, isolated, and identified from cattle in Beijing, China, during herd screening by BVDV antigen-ELISA and indirect immunofluorescence assay (IFA). To investigate its genomic features, the characteristic 5'UTR region of the isolates were sequenced and BLAST analyzed. BVDV BJ175170 belongs to the BVDV-1c subtype, which differs from the Beijing prevalent BVDV strains. The BVDV particles were further observed by transmission electron microscopy (TEM). To evaluate the virulence of the BVDV BJ175170, the BVDV seronegative rabbits were intraperitoneally inoculated with the virus suspension. Blood samples were analyzed for changes in leukocyte number and antibody titer, and tissue samples were taken for histopathology analysis. These data confirmed again that rabbits could act as the reservoir of BVDV, which poses a small but non-zero risk of re-infection for BVDV-free cattle herds. To our knowledge, this is the first report of pathological changes in rabbits after exposure to BVDV-1c subtype, which could act as experimental reference. Meanwhile, the results of this study indicate that rabbits could act as a potential model for studying the mechanism of BVDV in vivo.

Forsythiaside A Improves the Inhibitory Efficiency of Recombinant Protein Vaccines against Bovine Viral Diarrhea Virus Infection.

Bovine viral diarrhea virus (BVDV) is a critical animal pathogen that leads to cattle production losses associated with acute disease, immune dysregulation, reproductive failure, and respiratory disease. Due to the monotonous control technique and neglect of BVDV, increasing prevalence of BVDV has caused significant economic losses in the cattle industry worldwide. Therefore, novel anti-BVDV drugs are essential to prevent and control BVDV. Our previous studies have found that Forsythoside A (FTA) could inhibit the replication of BVDV via TRAF2-dependent CD28-4-1BB signaling in bovine peripheral blood mononuclear cells (PBMCs), but whether they can directly inhibit the BVDV remains unclear. Here, we further investigated the effects of FTA on BVDV and its underlying mechanisms of action. We found that FTA significantly inhibited the replication of BVDV in the MDBK cell directly. The results demonstrated that FTA could reduce the functional activation of Caspase-1 to inhibit the inflammatory response caused by BVDV infection and increase the expression of type I interferon (IFN-I) to clear the virus in vitro. The animal experiment was performed to evaluate the antiviral effect of FTA in vivo. Notably, after challenged with BVDV, mice with FTA + Erns-E2 protein displayed alleviated pathological damage and decreased the viral load in the spleen compared with mice inoculated with Erns-E2 protein. Furthermore, treatment with FTA enhanced body defense and delayed infection by the BVDV. Our results reveal that FTA suppresses BVDV replication both in vitro and in vivo and therefore shows promise as an anti-BVDV agent.

Z. morio Hemolymph Relieves E. coli-Induced Mastitis by Inhibiting Inflammatory Response and Repairing the Blood-Milk Barrier.

Escherichia coli (E. coli) is a major environmental pathogen causing coliform mastitis, characterized by cell death and mammary tissue damage. Our previous study has shown the antimicrobial effect of Zophobas morio (Z. morio) hemolymph against mastitis pathogens. In this study, we established E. coli-induced cellular and animal models for mastitis, aiming to evaluate the protective effect of Z. morio hemolymph against E. coli-induced mastitis in vivo and in vitro. In mice with E. coli, Z. morio hemolymph attenuated bacterial burden and histopathological impairment, reduced the production of interleukin (IL)-1ß, IL-18, tumor necrosis factor-α (TNF-α) and the ratio of CD4+ T/CD8+ T, and increased the production of IL-2 triggered by E. coli. Z. morio hemolymph also enhanced the integrity of the blood-milk barrier in E. coli-induced mastitis. In E. coli-stimulated porcine mammary epithelial cells, Z. morio hemolymph inhibited E. coli-induced inflammatory responses and upregulated tight junction proteins (ZO-1, Claudin-3 and Occludin). Moreover, we found that the anti-inflammatory effect of Z. morio hemolymph was mediated by inhibiting E. coli-induced NLRP3 inflammasome assembly, Caspase-1 activation, and reversing the inhibitory effect of E. coli on autophagy. Besides, Z. morio hemolymph augmented ATG5/ATG16L1-mediated autophagy activation, negatively regulated NLRP3 inflammasome activation. Our results reveal that Z. morio hemolymph alleviates E. coli-induced mastitis via lessening the inflammatory response by regulating the NLRP3 and ATG5/ATG16L1 signaling pathway, as well as repairing the blood-milk barrier.

CO2 Footprint of Thermal Versus Photothermal CO2 Catalysis.

Transformation of CO2 into value-added products via photothermal catalysis has become an increasingly popular route to help ameliorate the energy and environmental crisis derived from the continuing use of fossil fuels, as it can integrate light into well-established thermocatalysis processes. The question however remains whether negative CO2 emission could be achieved through photothermal catalytic reactions performed in facilities driven by electricity mainly derived from fossil energy. Herein, we propose universal equations that describe net CO2 emissions generated from operating thermocatalysis and photothermal reverse water-gas shift (RWGS) and Sabatier processes for batch and flow reactors. With these reactions as archetype model systems, the factors that will determine the final amount of effluent CO2 can be determined. The results of this study could provide useful guidelines for the future development of photothermal catalytic systems for CO2 reduction.

Sulphur-Embedded Hydrocarbon Belts: Synthesis, Structure and Redox Chemistry of Cyclothianthrenes.

Cyclothianthrenes, a series of sulphur-embedded hydrocarbon belts proposed a decade ago, were successfully constructed through a stepwise bottom-up synthesis. The belt [6]cyclothianthrene ([6]CT) is the smallest and most strained member of the family yet reported. Both [6]CT and [8]CT are the first examples of cyclothianthrene characterized by single crystal X-ray diffraction. An unprecedented chiral belt [7]CT and a Möbius-shaped [9]CT were also achieved via modular synthesis. Crystallographic and computational studies show that belts [6]CT-[8]CT have prism-like conformations with well-defined tubular cavities which have potential for guest molecule inclusion. Cyclic voltammograms further revealed that these belts are redox-active. The success of constructing sulphur-embedded hydrocarbon belts, that is, cyclothianthrenes, greatly enriches the chemistry of heteroatom-doped molecular belts and tubes.

Design of magnetic nanoparticles with high magnetic separation efficiencies and durability for Cu2+ adsorption.

Fe3O4@SiO2 core-shell structured nanoparticles are promising candidates for adsorption of heavy metal ions from waste water due to their high adsorption capacities and feasible recycling process. However, their practical applications have been hindered by the trade-off between the magnetic separation efficiency and durability. In this study, we demonstrate the preparation of amino-functionalized Fe3O4@SiO2 absorbents with both high magnetic separation efficiencies and durability in strong acidic environment. Our key strategy was to use a thin but highly protective silica layer by manipulating the sol-gel chemistry. With the protection of a thin but poorly permeable silica shell, the durability of the Fe3O4 in strong acid solution was significantly enhanced while its magnetic separation efficiency was maintained. We also demonstrated the adsorption of Cu2+ from aqueous solution corresponded well with the pseudo-second-order kinetic model and the Langmuir adsorption isotherm. The adsorption capacity can be further improved by coating an additional layer of mesoporous SiO2 outside Fe3O4@SiO2. The design of highly durable magnetic absorbents without sacrificing the magnetic separation efficiency greatly facilitates the practical applications of magnetic nanoparticles in adsorption of heavy metal ions.

Optimization-Based Wi-Fi Radio Map Construction for Indoor Positioning Using Only Smart Phones.

Fingerprinting-based Wi-Fi indoor positioning has great potential for positioning in GPS-denied areas. However, establishing a fingerprinting map (also called a radio map) prior to positioning (site survey) is normally a labor-intensive task. This paper proposes a method for easy site survey without need for any extra hardware. The user can conduct the site survey adopting only a smart phone. The collected inertial-based readings are processed using the pedestrian dead-reckoning algorithms to generate a raw trajectory. Then a factor graph optimization method is proposed to re-estimate the trajectory by adding constraints originated from collected Wi-Fi fingerprints and landmark positions. The proposed method is verified through an experiment in a mall. The mean positioning error is 1.10 m and the maximum error is 2.25 m. This level of positioning accuracy is considered sufficient for radio map generation purposes. A classical baseline algorithm, the k-Nearest Neighbor (kNN) algorithm, is adopted to test the positioning performance of the radio map (RM), which also validates the quality of the constructed RM from the proposed method.

Target Recognition of SAR Images via Matching Attributed Scattering Centers with Binary Target Region.

A target recognition method of synthetic aperture radar (SAR) images is proposed via matching attributed scattering centers (ASCs) to binary target regions. The ASCs extracted from the test image are predicted as binary regions. In detail, each ASC is first transformed to the image domain based on the ASC model. Afterwards, the resulting image is converted to a binary region segmented by a global threshold. All the predicted binary regions of individual ASCs from the test sample are mapped to the binary target regions of the corresponding templates. Then, the matched regions are evaluated by three scores which are combined as a similarity measure via the score-level fusion. In the classification stage, the target label of the test sample is determined according to the fused similarities. The proposed region matching method avoids the conventional ASC matching problem, which involves the assignment of ASC sets. In addition, the predicted regions are more robust than the point features. The Moving and Stationary Target Acquisition and Recognition (MSTAR) dataset is used for performance evaluation in the experiments. According to the experimental results, the method in this study outperforms some traditional methods reported in the literature under several different operating conditions. Under the standard operating condition (SOC), the proposed method achieves very good performance, with an average recognition rate of 98.34%, which is higher than the traditional methods. Moreover, the robustness of the proposed method is also superior to the traditional methods under different extended operating conditions (EOCs), including configuration variants, large depression angle variation, noise contamination, and partial occlusion.

Molecular Recognition and Chirality Sensing of Epoxides in Water Using Endo-Functionalized Molecular Tubes.

Chiral epoxides are important intermediates in chemistry and biology. The high-throughput screening of asymmetric epoxidation conditions requires fast determination of the absolute configurations and ee values of chiral epoxides. Herein, we report molecular recognition and chiroptical sensing of epoxides in water using endo-functionalized molecular tubes. The absolute configurations and ee values were simultaneously determined by circular dichroism spectroscopy. In addition, real-time monitoring as well as the application to real asymmetric epoxidation was demonstrated. The method is simple, environmentally friendly, and amenable to high-throughput screening.

Nanosized Pd-Au bimetallic phases on carbon nanotubes for selective phenylacetylene hydrogenation.

Palladium (Pd)-catalyzed selective hydrogenation of alkynes has been one of the most studied hydrogenation reactions in the last century. However, kinetic studies conducted to reveal the catalyst's active centers have been hindered because of dynamic surface changes on Pd during the reaction. In the present study, bimetallic Pd-Au nanoparticles supported on carbon nanotubes have been synthesized at room temperature as catalysts for selective hydrogenation of phenylacetylene, which show effectively enhanced selectivity compared to their monometallic counterparts. Structural and surface analyses of fresh and reacted catalysts reveal that selective hydrogenation of phenylacetylene is favored over nanosized Pd-Au bimetallic phases due to modifications in the Pd surface in terms of neighboring site isolation and electron density reduction.

Response of alternative splice isoforms of OsRad9 gene from Oryza sativa to environmental stress.

Rad9 protein plays an important role in cell-cycle checkpoint signal transduction in human and yeast cells, but knowledge about Rad9 in plants is limited. This study reports that the Rad9 gene of rice can generate the transcript products OsRad9.1 and OsRad9.2 through alternative splicing. OsRad9.1, with all nine exons, is the main cell-cycle checkpoint protein involved in the response of rice to genotoxic stresses (ultraviolet radiation and antibiotic stress), environmental stresses (drought, salt, and heavy metal stress), and auxin stimuli (2,4-D, IAA, and IBA). Meanwhile, transcript isoform OsRad9.2, which lost exon7 and exon8, showed different preferential stimulation effects on these stresses and pollen development duration. These results might indicat that besides the monitoring and repair of DNA damage, Rad9 might involve in the development of pollen.

Selective Recognition of Highly Hydrophilic Molecules in Water by Endo-Functionalized Molecular Tubes.

Selective recognition of neutral hydrophilic molecules in water is a challenge for supramolecular chemistry but commonplace in nature. By mimicking the binding pocket of natural receptors, endo-functionalized molecular tubes are proposed to meet this challenge. We found that two molecular tubes with inwardly directed hydrogen-bond donors recognize highly hydrophilic solvent molecules in water with high selectivity. In the complexes, hydrogen bonding occurs in the deep and hydrophobic cavity. The cooperative action between hydrogen bonding and hydrophobic effects accounts for the high affinity and selectivity. The molecular receptor is fluorescent and can detect concentrations of 1,4-dioxane-a known carcinogen and persistent environmental contaminant-in water at a limit of 119 ppb. The method simplifies the analytic procedure for this highly hydrophilic molecule.

GDP-D-mannose pyrophosphorylase from Pogonatherum paniceum enhances salinity and drought tolerance of transgenic tobacco.

Pogonatherum paniceum is a highly drought- and salt-tolerant plant species that is typically used for ecological restoration and the conservation of soil and water in many countries. Understanding the molecular mechanisms underlying plant abiotic stress responses, especially to salinity and drought stresses, in species such as P. paniceum could be important to broader crop improvement efforts. GDP-D-mannose pyrophosphorylase (GMPase) is the limiting enzyme in the synthesis of L-ascorbic acid (AsA), which plays a crucial role in the detoxification of reactive oxygen species (ROS). We have cloned and characterized the cDNA of the PpGMP gene of P. paniceum encoding a GMPase. The full-length cDNA sequence contains 1411 nucleotides encoding a putative protein with 361 amino acid residues and an approximate molecular mass of 39.68 kDa. The GMPase transcript was up-regulated in P. paniceum plants subjected to salinity and drought stress, respectively. Transgenic tobacco expressing PpGMPase exhibited enhanced salinity and drought resistance, a higher seed germination rate, better growth performance, a higher AsA content, a more stable redox state, higher superoxide dismutase (SOD) activity, and lower levels of malonaldehyde (MDA) and H2O2 under drought and salinity stress. Taken together, expression of PpGMPase in tobacco conferred salinity and drought stress tolerance by increasing the content of AsA, thereby enhancing ROS-detoxifying functions. Thus, PpGMP is a potential candidate gene for crop improvement.

Liquid Hybridization and Solid Phase Detection: A Highly Sensitive and Accurate Strategy for MicroRNA Detection in Plants and Animals.

MicroRNAs (miRNAs) play important roles in nearly every aspect of biology, including physiological, biochemical, developmental and pathological processes. Therefore, a highly sensitive and accurate method of detection of miRNAs has great potential in research on theory and application, such as the clinical approach to medicine, animal and plant production, as well as stress response. Here, we report a strategic method to detect miRNAs from multicellular organisms, which mainly includes liquid hybridization and solid phase detection (LHSPD); it has been verified in various species and is much more sensitive than traditional biotin-labeled Northern blots. By using this strategy and chemiluminescent detection with digoxigenin (DIG)-labeled or biotin-labeled oligonucleotide probes, as low as 0.01-0.25 fmol [for DIG-CDP Star (disodium2-chloro-5-(4-methoxyspiro{1,2-dioxetane-3,2'-(5'-chloro)tricyclo[3.3.1.13,7]decan}-4-yl)phenyl phosphate) system], 0.005-0.1 fmol (for biotin-CDP Star system), or 0.05-0.5 fmol (for biotin-luminol system) of miRNA can be detected and one-base difference can be distinguished between miRNA sequences. Moreover, LHSPD performed very well in the quantitative analysis of miRNAs, and the whole process can be completed within about 9 h. The strategy of LHSPD provides an effective solution for rapid, accurate, and sensitive detection and quantitative analysis of miRNAs in plants and animals.

Resveratrol attenuates inflammation and oxidative stress in epididymal white adipose tissue: implications for its involvement in improving steroidogenesis in diet-induced obese mice.

Chronic, low-grade systemic inflammation has been shown to play an important role in the development of obesity-related complications. Epididymal white adipose tissue (WAT) can influence testicular function through its endocrine function. The purpose of this study was to assess the effects of resveratrol on the epididymal WAT inflammatory response and on testicular steroidogenesis in obese individuals. Seven-week-old male C57BL/6J mice were fed a high-calorie and high-cholesterol diet (HCD group) or HCD supplemented with resveratrol (HCD+Res group) for 18 weeks. As we previously showed that resveratrol protects against Leydig cell steroidogenesis in HCD-induced obese mice, this study assessed macrophage infiltration in fat depots by measuring crown-like structure (CLS) density. Histological analysis showed that adipocyte size was significantly smaller and CLSs were less numerous in the HCD+Res group than the HCD group (P < 0.01). Additionally, resveratrol supplementation decreased Nfkb1 expression (P < 0.01) and increased the IκB-α protein abundance (P < 0.01) in epididymal WAT. Consistent with this alteration in NF-κB signaling, the expression of two classic proinflammatory cytokines, TNF-α (Tnfa) and IL-1ß (Il1b), were significantly decreased in the HCD+Res group compared with the HCD group (P < 0.01). Significant differences were also found in the expression of sirtuin1 (Sirt1) (P < 0.01) and manganese superoxide dismutase (Sod2) (P < 0.01) between the HCD and HCD+Res groups. Our data suggest that resveratrol can attenuate obesity-induced inflammation and oxidative stress in epididymal WAT, which partly accounts for its beneficial effects in testicular steroidogenesis.

A simple solution hybridization rapidly detects DNA by fluorescent probe.

Traditional Southern blot, as the golden standard to detect DNA, is widely used in molecular biology. However, its operation process has some significant disadvantages, such as radioactive contamination, complex procedures, time-consuming. Some new methods developed in recent years need expensive equipments and also have the defects of complex procedures and time-consuming. The paper described a method to detect DNA quickly by liquid hybridization. The probe used is a fragment of DNA sequence labeled by FITC-12-dUTP. The whole process includes four steps: probe preparation, hybridization, electrophoresis and signal detection. The comparative experiments using the single strand DNA probe and the double strand DNA probe indicated that the sensitivity of single strand DNA probes, which could detect 0.38 µg (1.82 x 10(-13) mol) plasmid, is 2.1 times than that of the double strand DNA probes which could detect 0.8 µg (3.64 x 10(-13) mol) plasmid. The total procedure is simple and can be completed in 3 h.

Chemically exfoliated boron nanosheets for efficient oxidative dehydrogenation of propane.

Oxidative dehydrogenation of propane (ODHP) is a promising technique for producing propene due to its low operative temperature and coke-resistant feature. Recently, boron-based catalysts have been widely investigated for ODHP owing to their brilliant performance. Herein, we report that boron in the form of nanosheets can be prepared feasibly by exfoliating layered MgB2 with hydrochloric acid, and can efficiently and stably catalyze ODHP. At 530 °C, the catalyst exhibits propene and ethene selectivities as high as 63.5% and 18.4%, respectively, at a 40% propane conversion. The olefin productivity reaches 2.48 golefin gcat-1 h-1, superior to the commercial h-BN and other reported boron-based catalysts. Even after testing for 100 h at 530 °C, the catalyst still maintains excellent stability. This work expands the effective boron-based catalyst family for ODHP and demonstrates the great potential of the new type of 2D material-boron nanosheet for energy and catalytic applications.