Structures of Foot-and-Mouth Disease Virus with Bovine Neutralizing Antibodies Reveal the Determinant of Intraserotype Cross-Neutralization.

Foot-and-mouth disease virus (FMDV) exhibits broad antigenic diversity with poor intraserotype cross-neutralizing activity. Studies of the determinant involved in this diversity are essential for the development of broadly protective vaccines. In this work, we isolated a bovine antibody, designated R55, that displays cross-reaction with both FMDV A/AF/72 (hereafter named FMDV-AAF) and FMDV A/WH/09 (hereafter named FMDV-AWH) but only has a neutralizing effect on FMDV-AWH. Near-atomic resolution structures of FMDV-AAF-R55 and FMDV-AWH-R55 show that R55 engages the capsids of both FMDV-AAF and FMDV-AWH near the icosahedral 3-fold axis and binds to the ßB and BC/HI-loops of VP2 and to the B-B knob of VP3. The common interaction residues are highly conserved, which is the major determinant for cross-reaction with both FMDV-AAF and FMDV-AWH. In addition, the cryo-EM structure of the FMDV-AWH-R55 complex also shows that R55 binds to VP3E70 located at the VP3 BC-loop in an adjacent pentamer, which enhances the acid and thermal stabilities of the viral capsid. This may prevent capsid dissociation and genome release into host cells, eventually leading to neutralization of the viral infection. In contrast, R55 binds only to the FMDV-AAF capsid within one pentamer due to the VP3E70G variation, which neither enhances capsid stability nor neutralizes FMDV-AAF infection. The VP3E70G mutation is the major determinant involved in the neutralizing differences between FMDV-AWH and FMDV-AAF. The crucial amino acid VP3E70 is a key component of the neutralizing epitopes, which may aid in the development of broadly protective vaccines. IMPORTANCE Foot-and-mouth disease virus (FMDV) causes a highly contagious and economically devastating disease in cloven-hoofed animals, and neutralizing antibodies play critical roles in the defense against viral infections. Here, we isolated a bovine antibody (R55) using the single B cell antibody isolation technique. Enzyme-linked immunosorbent assays (ELISA) and virus neutralization tests (VNT) showed that R55 displays cross-reactions with both FMDV-AWH and FMDV-AAF but only has a neutralizing effect on FMDV-AWH. Cryo-EM structures, fluorescence-based thermal stability assays and acid stability assays showed that R55 engages the capsid of FMDV-AWH near the icosahedral 3-fold axis and informs an interpentamer epitope, which overstabilizes virions to hinder capsid dissociation to release the genome, eventually leading to neutralization of viral infection. The crucial amino acid VP3E70 forms a key component of neutralizing epitopes, and the determination of the VP3E70G mutation involved in the neutralizing differences between FMDV-AWH and FMDV-AAF could aid in the development of broadly protective vaccines.

Eco-Corona vs Protein Corona: Effects of Humic Substances on Corona Formation and Nanoplastic Particle Toxicity in Daphnia magna.

Despite many studies on the toxicity of nanoplastic particles (NPPs) to aquatic invertebrates, the effects of ecological constituents such as humic substances (HSs) are often neglected. In our study, Daphnia magna was used to evaluate the effects of three HSs, natural organic matter (NOM), fulvic acid (FA), and humic acid (HA), on NPP toxicity and corona formation. Acute toxicities of NPPs were reduced by all HSs at environmentally relevant concentrations. NPPs elicited the upregulation of all genes related to detoxification, oxidative stress, and endocrine activity after 7 days of exposure. The presence of NOM or HA resulted in the mitigation of gene expression, whereas significantly higher upregulation of all of the genes was observed with FA. The presence of FA led to increased protein adsorption on NPPs in D. magna culture medium (eco-corona, EC) and homogenates (protein corona, PC), while there was less adsorption in the presence of HA. The highly abundant proteins identified in EC are involved in immune defense, cell maintenance, and antipredator response, while those in PC are responsible for lipid transport, antioxidant effects, and estrogen mediation. Our findings revealed the key influence of HSs on the toxicity of NPPs and provide an analytical and conceptual foundation for future study.

Spatial distribution of phthalate esters and the associated response of enzyme activities and microbial community composition in typical plastic-shed vegetable soils in China.

The widespread use of phthalate esters (PAEs) in plastic products has made them ubiquitous in environment. In this study, 93 soil samples were collected in 31 plastic-sheds from one of China's largest vegetable production bases, Shouguang City, Shandong Province, to investigate the pollution characteristics and composition of PAEs in soils. Eleven PAEs were detected in the soil samples with the total concentration of 756-1590 µg kg-1 dry soil. Di (2-ethylhexyl) phthalate (DEHP), bis (2-n-butoxyethyl) phthalate (DBEP), di-isobutyl phthalate (DiBP) and di-n-butyl phthalate (DBP) were the main pollutants with the highest concentrations. Moreover, soil properties, including pH, total organic carbon (TOC), soil enzyme activities, and soil microbial community characteristics, were monitored to explore the associated formation mechanisms. The concentration of PAEs in the plastic-shed vegetable soils was regionalized and the contamination degree in different regions was related to soil microbial characteristics and soil enzyme activities. Phthalate ester is positively correlated with catalase and sucrase, and negatively correlated with dehydrogenase and urease. Furthermore, some tolerant and sensitive bacteria were selected, which possibly could be used as potential indicators of PAE contamination in soil. Dimethyl phthalate (DMP) and DBP also had greater effects on the soil microbial community than other PAEs. The results will provide essential data and support the control of PAEs in plastic-shed vegetable soils in China.

Multifunctional Polyethylene Glycol (PEG)-Poly (Lactic-Co-Glycolic Acid) (PLGA)-Based Nanoparticles Loading Doxorubicin and Tetrahydrocurcumin for Combined Chemoradiotherapy of Glioma.

BACKGROUND This study aimed to prepare doxorubicin- and tetrahydrocurcumin-loaded and transferrin-modified PEG-PLGA nanoparticles (Tf-NPs-DOX-THC) for enhanced and synergistic chemoradiotherapy. MATERIAL AND METHODS Tf-NPs-DOX-THC were prepared via the double-emulsion method. The morphologies and particle sizes of the prepared nanoparticles were examined by TEM and DLS, respectively. The in vitro MTT, apoptosis, and clone formation assays were performed to detect the proliferation and radiosensitivity of cells with various treatments. Cellular uptake assay was also conducted. The tissue distribution of Tf-NPs was investigated by ex vivo DOX fluorescence imaging. The in vivo tumor growth inhibition efficiency of various treatments was evaluated in orthotopic C6 mouse models and C6 subcutaneously grafted mouse models. RESULTS Tf-NPs-DOX-THC exhibited high drug-loading efficiency (6.56±0.32%) and desirable particle size (under 250 nm). MTT, apoptosis, and clone formation assays revealed the enhanced anti-cancer activity and favorable radiosensitizing effect of Tf-NPs-DOX-THC. Strong fluorescence was observed in the brains of mice treated with Tf-NPs-DOX. The in vitro release of drug from nanoparticles was in a pH-sensitive manner. Tf-NPs-DOX-THC in combination with radiation also achieved favorable anti-tumor efficacy in vivo. CONCLUSIONS All results suggest that a combination of Tf-NPs-DOX-THC and radiation is a promising strategy for synergistic and sensitizing chemoradiotherapy of glioma.

The permeability of puerarin loaded poly(butylcyanoacrylate) nanoparticles coated with polysorbate 80 on the blood-brain barrier and its protective effect against cerebral ischemia/reperfusion injury.

Puerarin (PUE) is a good candidate for treating stroke, but its low concentration in brain after administration limits its curative efficacy. The aim of the present work was to design and characterize PUE loaded poly(butylcyanoacrylate) nanoparticles (PBCN) coated with polysorbate 80 (Ps 80), and to evaluate the effect of PBCN on the permeability of PUE across the blood-brain barrier (BBB) and the effect of PUE loaded PBCN on the cerebral ischemia/reperfusion injury. PUE loaded PBCN were successfully prepared by anionic polymerization method with the mean particle size of 201.2 nm and the zeta potential of -7.72 mV. The in vitro release behavior of PUE from the nanoparticles showed a biphasic profile manner with an initial burst release followed by a sustained release. The results of pharmacokinetic and biodistribution to brain performed in mice after intravenous administration showed that the drug concentrations in blood and brain for PUE loaded PBCN were both greater than these for the free drug. Moreover, compared with free drug, the vein injection of PUE loaded PBCN exerted the better neuroprotective effect in rats with focal cerebral ischemic injury via significantly decreasing neurological deficit scores, increasing body weight, lowing brain water content, and reducing the infarct volume. The results indicated that this preparation may reduce the total dose required for the stroke therapy with concurrent reduction in dose related toxicity. All these findings suggest that PBCN could enhance the transport of PUE to brain and have a potential as a neuroprotective agent in the focal cerebral ischemic injury.

Microplastics degradation stimulated by in-situ bioelectric field in agricultural soils.

Bioelectric field is a stimulated force to degrade xenobiotic pollutants in soils. However, the effect of bioelectric field on microplastics (MPs) aging is unclear. The degradation behavior of polyvinyl chloride (PVC), polyethylene (PE) and polylactic acid (PLA) was investigated in an agricultural soil microbial electrochemical system in which bioelectric field was generated in-situ by native microbes. Based on the density function theory, the energy gaps between the highest and the lowest occupied molecular orbitals of the three polymers with periodic structure were 4.20, 7.24 and 10.09 eV respectively, and further decreased under the electric field, indicating the higher hydrolysis potential of PLA. Meanwhile, the mass loss of PLA in the closed-circuit group (CC) was the highest on day 120, reaching 8.94%, which was 3.01-3.54 times of that without bioelectric field stimulation. This was mainly due to the enrichment of plastic-degrading bacteria and a robust co-occurrence network as the deterministic assembly process, e.g., the abundance of potential plastic-degrading bacteria on the surface of PLA and PVC in the CC increased by 1.92 and 1.30 times, respectively, compared to the open-circuit group. In terms of functional genes, the xenobiotic biodegradation and metabolism capacity of plasticsphere in the CC were stronger than that in soil, and determined by the bioaccessibility of soil nitrogen and carbon. Overall, this study explored the promoting effect of bioelectric field on the degradation of MPs and reveled the mechanism from quantum chemical calculations and microbial community analysis, which provides a novel perception to the in-situ degradation of MPs.

Development of a double-antibody sandwich ELISA for rapidly quantitative detection of residual non-structural proteins in inactivated foot-and-mouth disease virus vaccines.

Foot-and-mouth disease (FMD) is a highly contagious and economically devastating disease of cloven-hoofed animals. Vaccination and surveillance against non-structure protein (NSP) are the most efficacious and cost-effective strategy to control this disease. Therefore, vaccine purity control is vital for successful prevention. Currently, vaccine purity is tested by an in-vivo test that recommended in the World Organization for Animal Health (WOAH), but it is time consuming and costly. Herein, we develop a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for quantitative detection of residual NSPs in inactivated FMD virus (FMDV) vaccines. In this assay, the monoclonal antibody 3A24 was selected as capture antibody and biotinylated 3B4B1 (Biotin-3B4B1) as detection antibody. A standard curve was developed using the NSP 3AB concentration versus OD value with the linear range of concentration of 2.5-160 ng/mL. The lowest limit of detection was 2.5 ng/mL. In addition, we determined 2.5 ng/mL of NSP as an acceptable threshold value of FMD vaccine purity using a dose-response experiment in cattle. The DAS-ELISA combined with the threshold value of FMD vaccine purity could provide a quick and simple tool for evaluation the antigenic purity of FMD vaccine during the manufacturing process.

Dynamic control of pentachlorophenol photodegradation process using P25/PDA/BiOBr through regulation of photo-induced active substances and chemiluminescence.

Photodegradation is a new approach for the removal of pentachlorophenol (PCP). Photooxidation degradation (using hydroxyl radicals) exhibits better performance to remove PCP than photoreduction degradation, but the former will lead to an increase in the production of toxic by-products such as tetrachloro-1,4-benzoquinone (TCBQ). Thus, a new strategy is required to enhance PCP photodegradation and simultaneously inhibit toxic intermediates production. Herein, TiO2 (P25)/polydopamine (PDA)/BiOBr was synthesized and used to photodegrade PCP. Based on the relative position of the CB and VB of P25 and BiOBr, and PDA as an electron transfer mediator, a high number of holes, electrons, and superoxide anions were produced instead of hydroxyl radicals. The photocatalytic activity of P25/PDA/BiOBr exhibited the best performance among as-prepared samples, reaching a k(pcp) value of 0.4 min-1 (20 µM PCP) under UV light irradiation within 10 min. According to chemiluminescence and acute toxicity assays, relative to P25, the toxic intermediates of TCBQ and trichlorohydroxy-1,4-benzoquinone (OH-TrCBQ) generation was greatly reduced over P25/PDA/BiOBr, with a lack of toxic product generation during PCP photodegradation process. These findings provide an alternative strategy to achieve greener and more efficient organic pollutant photodegradation.

Natural polymeric nanocarriers in malignant glioma drug delivery and targeting.

Among all central nervous diseases, malignant glioma is a crucial part that deserves more attention since high fatality and disability rate. There are several therapeutic strategies applied to the treatment of malignant glioma, especially certain chemotherapy-related treatments. However, the existence of the blood-brain barrier (BBB) seriously hinders the strategy's progress, so how to escape from the barriers is a fascinating question. Herein, we comprehensively discussed the details of malignant glioma and the BBB's functional morphology and summarised several routes bypassing the BBB. Additionally, since possessing excellent properties for drug delivery, we provided an insight into various promising natural polymeric materials and highlighted their applications in the treatment of malignant glioma.

Electrospun Polymer-Free Nanofibers Incorporating Hydroxypropyl-ß-cyclodextrin/Difenoconazole via Supramolecular Assembly for Antifungal Activity.

In this study, flexible and self-standing hydroxypropyl-ß-cyclodextrin/difenoconazole inclusion complex (HPßCD/DZ-IC) nanofibers were prepared by polymer-free electrospinning, which exhibited potential to be a new fast-dissolving pesticide formulation. Scanning electron microscopy and optical microscopy were applied to evaluate the morphology of nanofibers, which showed that the resulting HPßCD/DZ-IC nanofibers were bead-free and uniform. In addition, the proton nuclear magnetic resonance (1H NMR) spectrum suggested a stoichiometric ratio of 1:0.9 (HPßCD/DZ). Other characterization methods, such as UV-vis absorption, fluorescence spectroscopy, Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and thermogravimetric analysis (TGA), were applied in this study. On the one hand, UV-vis absorption, fluorescence spectroscopy, FT-IR, XRD, and TGA provided useful information for the successful formation of an inclusion complex; on the other hand, the results of TGA indicated the thermal stability of DZ was enhanced after the formation of inclusion complexes. Besides, the phase solubility test could explain the increased water solubility of the nanofibers of inclusion complexes formed by DZ and HPßCD. The results of molecular docking studies demonstrated the most favorable binding interactions when HPßCD combined with DZ. The dissolution test and the antifungal performance test exhibited the characteristics of fast dissolution and the excellent antifungal performance of HPßCD/DZ-IC nanofibers, respectively.

Microplastics from consumer plastic food containers: Are we consuming it?

Microplastic (MP) accumulation in the environment has become an issue of human and environmental importance. Great efforts were made recently to identify the sources of MP exposure to humans and their release into the environment. Here, we employed spectroscopic techniques to identify and characterize MP in consumer plastic food containers that are, in huge quantity, used for food delivery and disposable plastic cups for daily drinking. We determined the average weight of isolated MP per pack to be 12 ± 5.12 mg, 38 ± 5.29 mg, and 3 ± 1.13 mg for the round-shaped, rectangular-shaped plastic container and disposable plastic cups, respectively, with various morphological features including cubic, spherical, rod-like as well as irregular shapes, which may either be consumed by humans or released into the environment. This study demonstrates that new plastic containers can be an important source of direct human and environmental exposure to microplastics. Most importantly, our results indicated that necessary attention must be given to morphological features of realistic MPs when evaluating their risks to humans and the environment.

Encapsulation of thiabendazole in hydroxypropyl-ß-cyclodextrin nanofibers via polymer-free electrospinning and its characterization.

BACKGROUND: Thiabendazole (TBZ) is a poorly water-soluble benzimidazole fungicide. However, the water solubility of TBZ can be significantly enhanced by inclusion complexation with cyclodextrins. In this study, a thiabendazole/hydroxypropyl-ß-cyclodextrin (TBZ/HPßCD) complex was synthesized and electrospinning was performed to produce a TBZ/HPßCD nanofibrous (TBZ/HPßCD-NF) complex that improved water solubility and antifungal activity. RESULTS: The formation of TBZ/HPßCD-NF was characterized by Fourier transform infrared spectroscopy, X-ray diffraction and nuclear magnetic resonance. The morphology of TBZ/HPßCD-NF was studied by scanning electron microscopy. A phase solubility experiment showed that HPßCD exerted a great solubilization effect on TBZ, and TBZ/HPßCD-NF had better antifungal activity compared to that of TBZ alone. CONCLUSIONS: In summary, the solid fungicidal nanodispersion prepared in the present study is a new type of formulation that can enhance the water solubility of TBZ. This formulation, which demonstrated potential as a new fast dissolving formulation type with increased efficacy, is expected to be conducive to the sustainable development of agriculture. © 2020 Society of Chemical Industry.

Comparative study on the pollution status of organochlorine pesticides (OCPs) and bacterial community diversity and structure between plastic shed and open-field soils from northern China.

The pollution status of organochlorine pesticides (OCPs) and microbial community in plastic shed and open-field soils may be different due to the significant variations in environmental factors between the two cultivation modes. However, the differences remain unclear. We conducted a regional-scale survey to investigate the pollution level, distribution, and sources of 20 OCPs, and to evaluate the soil physicochemical properties and bacterial community in soils from plastic shed and open-field locating the north areas of China. We found that levels of total OCPs in the plastic shed soils were significantly higher than those in the nearby open-field soils. Most of these OCPs were attributed to historical application, except for dichlorodiphenyltrichloroethanes (DDTs) due to the fresh input along with dicofol application. Soil pH (for both cultivation modes) and total organic carbon (TOC) content (only for plastic sheds) were significantly correlated with the total OCP concentrations. Additionally, microbial diversity and richness were generally lower in plastic shed soils than in nearby open-field soils for each region. The bacterial community variation among different regions might be principally determined by the soil type. Soil pH had the greatest impact on the microbial community across all plastic shed and open-field samples. These results provide a better understanding of the environmental impact and ecological risk of OCPs in soils with different cultivation modes.

Advances in liver-directed gene therapy for hepatocellular carcinoma by non-viral delivery systems.

Hepatocellular carcinoma (HCC) is a malignancy with a high mortality. Gene therapy provides a promising way for the treatment of HCC. Efficient gene delivery system, suitable gene target and appropriate way of administration together determine the effect of gene therapy for HCC. In recent years, employing non-viral gene delivery systems in gene therapy for HCC has attracted a lot of attention. Compared with viral vectors, non-viral gene delivery systems are nearly non-immunogenic, relatively safer, less expensive to produce and can carry a good many of genetic materials. But the transfection efficiency of these vectors still needs to be improved. And the liver targeting is another problem that needs to be solved. Attaching ligands to the non-viral vectors to enhance the targeting ability to the specific receptor and targeting to molecular targets of HCC are the effective strategies. Adopting suitable ways of administration is also a factor that plays an important role to achieve liver targeting. This review introduced the advances in liver-targeted gene therapy by non-viral vectors including the efforts to overcome the low transfection efficiency and enhance the liver targeting effect.

[Clinical application of self-made drainage tube with balloon for iatrogenic colonic perforation].

OBJECTIVE: To investigate the clinical efficacy of colonic bypass drainage by self-made drainage tube with balloon for iatrogenic colonic perforation. METHODS: A retrospective analysis of 8 patients with iatrogenic colonic perforations from January 2009 to March 2011 was performed. Self-made drainage tubes with balloon were placed in the bowel lumen endoscopically after perforations were closed with endoclips or endoloops under endoscope. The inflatable balloon at the front-end of the tube was fixed at the mouth side of colonic perforation to achieve continuous drainage of stool and intestinal juice. RESULTS: Endoscopic bypass continuous drainage by using self-made drainage tube with balloon was successfully carried out in all the 8 patients. All the perforations healed and no surgical intervention required. Bypass drainage continued for 3-10 days(mean 7.6 days). One patient received colonoscopy 3 days after the procedure, and displacement of the drainage tube was noticed requiring endoscopic adjustment. All the drainage tubes were removed uneventfully, and no ulceration or perforation occurred at balloon fixed site after removal. After follow up ranging from 12 to 36 months, no chronic fistula, adhesive obstruction, or abdominal infection occurred. CONCLUSION: Colonic bypass drainage by self-made drainage tube with balloon for iatrogenic colonic perforation is simple, feasible, safe and reliable.

CpG oligodeoxynucleotide and montanide ISA 206 adjuvant combination augments the immune responses of a recombinant FMDV vaccine in cattle.

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. To prevent the spread of FMDV, inactivated virus vaccines are used to immunize animals in developing countries. However, there are safety concerns. In addition, it is difficult to distinguish the vaccinated animals from those naturally infected ones. In our lab, we have developed a recombinant FMDV vaccine named A7. A7 contained multiple B cell and T cell epitopes, which reside in a capsid protein (VP1) of FMDV. To enhance its immunogenicity, A7 was formulated with CpG ODN RW03 in combination with Montanide ISA 206 (ISA), and the resultant vaccine (A7+ISA+CpG ODN RW03) was used to immunize mice and cattle. It was found that CpG ODN RW03 and ISA combination could facilitate A7 to induce a vigorous and long-lasting specific antibody response in mice and cattle. After FMDV challenge, 80% (4/5) of the calves immunized with A7+ISA+CpG ODN RW03 were protected, which was superior to those immunized with A7+ISA (25%, 1/4) or inactivated FMDV vaccine (50%, 2/4). These findings suggest that CpG ODN RW03 could be used with Montanide ISA 206 as a potent adjuvant for recombinant FMDV in cattle.

Molecularly imprinted polymer as micro-solid phase extraction combined with high performance liquid chromatography to determine phenolic compounds in environmental water samples.

The molecularly imprinted polymer with 2,4,6-trichlorophenol (2,4,6-TCP) as the template molecule and methylacrylic acid (MAA), divinylbenzene (DVB) as functional monomer and the crosslinker, respectively, has been prepared and used as molecularly imprinted micro-solid phase extraction (MIMSPE) procedure for the selectively preconcentration of phenolic compounds from environmental water samples. Various parameters for the extraction efficiency of the MIMSPE have been evaluated. The optimized MIMSPE method allowed the extraction of the analytes from the sample matrix followed by a selective washing step using acetonitrile containing 0.3% (v/v) acetic acid. The characteristics of the MIMSPE method were valid by high performance liquid chromatography. The recoveries ranged between 88.9% and 102.5% for tap water, between 80.0% and 94.0% for river water, between 80.0% and 90.5% for sewage water fortified with 0.4 mg L(-1) of phenol, 4-chlorophenol (4-CP), 2,4-dichlorophenol (2,4-DCP), 2,4,6-TCP, pentachlorophenol (PCP) were obtained. This method has been successfully applied to preconcentrate and determine of phenolic compounds in environmental water samples directly.

Molecularly imprinted solid-phase extraction combined with high performance liquid chromatography for analysis of phenolic compounds from environmental water samples.

The molecularly imprinted bulk polymer with 2,4,6-trichlorophenol (2,4,6-TCP) as the template molecule and methylacrylic acid (MAA), ethylene glycol dimethacrylate (EGDMA) as functional monomer and the crosslinker, respectively, has been prepared and applied to the molecularly imprinted solid-phase extraction (MISPE) procedure for selective preconcentration of phenolic compounds from environmental water samples. Various parameters affecting the extraction efficiency of the polymer have been evaluated to optimize the selective preconcentration of the phenolic compounds from aqueous samples. The characteristics of the MISPE method were validated by HPLC. The recoveries ranged between 90% and 98% (RSD: 0.9-2.3%, n=3) for tap water, between 85% and 105% (RSD: 2.6-4.9%, n=3) for river water, between 78% and 98% (RSD: 2.6-5.4%, n=3) for sewage water fortified with 0.4 mg L(-1) of phenol, 4-chlorophenol (4-CP), 2,4-dichlorophenol (2,4-DCP), pentachlorophenol (PCP). It was demonstrated that this MISPE-HPLC method could be applied to direct preconcentration and determination of phenolic compounds in environmental water samples.