GRP78 recognizes EV-F 3D protein and activates NF-κB to repress virus replication by interacting with CHUK/IKBKB.

Enteroviruses are the causative agents associated with several human and animal diseases, posing a significant threat to human and animal health. As one of the host immune defense strategies, innate immunity plays a crucial role in defending against invading pathogens, where the host utilizes a variety of mechanisms to inhibit or eliminate the pathogen. Here, we report a new strategy for the host to repress enterovirus replication by the 78 kDa glucose-regulated protein (GRP78), also known as heat shock protein family A member 5 (HSPA5). The GRP78 recognizes the EV-encoded RNA-dependent RNA polymerases (RdRPs) 3D protein and interacts with the nuclear factor kappa B kinase complex (CHUK) and subunit beta gene (IKBKB) to facilitate the phosphorylation and nuclear translocation of NF-κB, which induces the production of inflammatory factors and leads to a broad inhibition of enterovirus replication. These findings demonstrate a new role of GRP78 in regulating host innate immunity in response to viral infection and provide new insights into the mechanism underlying enterovirus replication and NF-κB activation.IMPORTANCEGRP78 is known as a molecular chaperone for protein folding and plays a critical role in maintaining protein folding and participating in cell proliferation, cell survival, apoptosis, and metabolism. However, the functions of GRP78 to participate in enterovirus genome replication and innate immune responses are rarely documented. In this study, we explored the functions of the EV-3D-interacting protein GRP78 and found that GRP78 inhibits enterovirus replication by activating NF-κB through binding to EV-F 3D and interacting with the NF-κB signaling molecules CHUK/IKBKB. This is the first report that GRP78 interacts with CHUK/IKBKB to activate the NF-κB signaling pathway, which leads to the expression of the proinflammatory cytokines and inhibition of enterovirus replication. These results demonstrate a unique mechanism of virus replication regulation by GRP78 and provide insights into the prevention and treatment of viral infections.

Augmented-reality based brain-computer interface of robot control.

Brain Computer Interface (BCI) is a new approach to human-computer interaction. It can control the external devices directly with the brain without words and body movements. Brain-controlled robot is a major research area in the field of BCI, which organically integrates BCI with robotic systems to achieve safe and effective real-time control of robots using the user's electroencephalogram (EEG). Currently, there are two types of control methods for brain-controlled robots. One is direct control and the other is shared control. Direct brain control has its shortcomings, namely, low control efficiency and easy user fatigue. Shared control technique can effectively improve the control of brain-controlled robots and reduce the thinking ability of brain-controlled robots, thus making it the main control method of brain-controlled robots. The brain-computer collaborative control system based on augmented reality (AR) technology studied in this paper is a human-computer shared control method. In the experimental analysis of virtual reality (VR) systems and AR systems, this paper processes polylines through a series of control vertices with specific coordinates, using the relative distance measured between each point and the starting point as the relative coordinates, and calculates the operational errors of the two types of systems. In the system error of machining broken lines, when the relative coordinates are (10, 20), (40, 50), and (70, 80), the error values of the VR system are 0.17 mm, 0.36 mm, and 0.55 mm, respectively, while the error values of the AR system are 0.11 mm, 0.24 mm, and 0.41 mm, respectively. Therefore, the studies have illustrated the importance of AR systems for the study of brain-computer collaborative control of robots.

Genetic diversity and recombination of bovine enterovirus strains in China.

Bovine enterovirus (BEV) consisting of enterovirus species E (EV-E) and F (EV-F) is the causative agent associated with respiratory and gastrointestinal diseases in cattle. Here, we reported the characterization, genetic diversity, and recombination of novel BEV strains isolated from the major cattle-raising regions in China during 2012-2018. Twenty-seven BEV strains were successfully isolated and characterized. Molecular characterization demonstrated that the majority of these novel BEV strains (24/27) were EV-E, while only few strains (3/27) were EV-F. Sequence analysis revealed the diversity of the circulating BEV strains such as species and subtypes where different species or subtype coinfections were detected in the same regions and even in the same cattle herds. For the EV-E, two novel subtypes, designated as EV-E6 and EV-E7, were revealed in addition to the currently reported EV-E1-EV-E5. Comparative genomic analysis revealed the intraspecies and interspecies genetic exchanges among BEV isolates. The representative strain HeN-B62 was probably from AN12 (EV-F7) and PS-87-Belfast (EV-F3) strains. The interspecies recombination between EV-E and EV-F was also discovered, where the EV-F7-AN12 might be from EV-E5 and EV-F1, and EV-E5-MexKSU/5 may be recombined from EV-F7 and EV-E1. The aforementioned results revealed the genetic diversity and recombination of novel BEV strains and unveiled the different BEV species or subtype infections in the same cattle herd, which will broaden the understanding of enterovirus genetic diversity, recombination, pathogenesis, and prevention of disease outbreaks. IMPORTANCE: Bovine enterovirus (BEV) infection is an emerging disease in China that is characterized by digestive, respiratory, and reproductive disorders. In this study, we first reported two novel EV-E subtypes detected in cattle herds in China, unveiled the coinfection of two enterovirus species (EV-E/EV-F) and different subtypes (EV-E2/EV-E7, EV-E1/EV-E7, and EV-E3/EV-E6) in the same cattle herds, and revealed the enterovirus genetic exchange in intraspecies and interspecies recombination. These results provide an important update of enterovirus prevalence and epidemiological aspects and contribute to a better understanding of enterovirus genetic diversity, evolution, and pathogenesis.

Investigating local field tuning Fermi resonance of CS2 by Raman spectroscopy and DFT calculations.

In the spectroscopic study of polyatomic molecules, Fermi resonance (FR) is a vibrational coupling and energy transfer phenomenon that widely exists intra- and intermolecular. In particular, the FR coupling between the fundamental mode ν1 and the doubling mode 2ν2 of the CS2 molecule has attracted extensive research. In this work, we investigate the effect of local field on tuning the FR of CS2. By analyzing the Raman spectra of CS2 mixed with methanol and ethanol with different mole fractions, the results indicated that weak HBs interactions in binary solutions can be reflected by the linear frequency shift of the C-H bond vibrations (in methanol and ethanol) with different molar concentrations. Furthermore, the geometrical structure was optimized using DFT simulation, and the vibration analysis and interaction energy were carried out. The simulated Raman spectra are in good agreement with the experiments. In addition, high-pressure Raman spectra of CS2 were obtained by diamond anvil cell technique (up to 9.19 GPa) and a pressure-induced phase transition was observed at 1.71 GPa. The results demonstrated that the pressure-induced polymerization phase transition of CS2 molecules causes the close packing and more orderly arrangement of molecules, resulting in the enhancement of FR coupling. HB and high pressure tune the FR of the CS2 molecule differently.

CACNN: Capsule Attention Convolutional Neural Networks for 3D Object Recognition.

Recently, view-based approaches, which recognize a 3D object through its projected 2-D images, have been extensively studied and have achieved considerable success in 3D object recognition. Nevertheless, most of them use a pooling operation to aggregate viewwise features, which usually leads to the visual information loss. To tackle this problem, we propose a novel layer called capsule attention layer (CAL) by using attention mechanism to fuse the features expressed by capsules. In detail, instead of dynamic routing algorithm, we use an attention module to transmit information from the lower level capsules to higher level capsules, which obviously improves the speed of capsule networks. In particular, the view pooling layer of multiview convolutional neural network (MVCNN) becomes a special case of our CAL when the trainable weights are chosen on some certain values. Furthermore, based on CAL, we propose a capsule attention convolutional neural network (CACNN) for 3D object recognition. Extensive experimental results on three benchmark datasets demonstrate the efficiency of our CACNN and show that it outperforms many state-of-the-art methods.

Pressure-induced Fermi resonance between fundamental modes in α-Hydroquinone.

Fermi resonance (FR), a prevalent phenomenon in molecules, has an important effect in spectrum analysis. As an effective way to change the molecular structure and tune symmetry, high-pressure techniques can often induce FR. Hydroquinone (HQ) is a hydrogen-bonded crystal that tends to form a solid inclusion compound with a suitable guest and has wide applications. Inthiswork, a high-pressure technique was used to investigate α-HQ using high pressure to tune the symmetry to produce FR. Raman and infrared spectra of α-HQ were investigated at ambient pressure, and then Raman spectra under high pressure of α-HQ were investigated up to 19.64 GPa. Results indicated that there were two phase transitions found at about 3.61 and 12.46 GPa. Fundamental FR was not present in α-HQ molecules at ambient pressure. At 3.61 GPa, the first-order phase transition occurred due to the pressure-induced symmetry change, resulting in two Raman modes at 831 cm-1 and 854 cm-1 with the same symmetry, thereby providing evidence that the fundamental FR phenomenon occurred. Furthermore, the pressure-induced changes of the FR parameters were elucidated. Thus pressure provided an effective way to study FR between two asymmetric species.

A Neonatal Murine Model for Caprine Enterovirus Infection and the Viral Tissue Tropism.

As the first caprine enterovirus identified from goat herds characterized by severe diarrhea with a high morbidity and mortality rate, the underlying pathogenesis and tissue tropism for CEV-JL14 remains largely unknown. Here, we reported the establishment of a neonatal murine model for caprine enterovirus and the unveiling of the tissue tropism and underlying pathogenesis for CEV-JL14 enterovirus. Susceptible murine strains, the infective dose, the infective routes, viral loads, and tissue tropism for CEV-JL14 infection were determined. The findings showed that ICR mice were susceptible to CEV-JL14 infection via all infection routes. Tissue viral load analysis showed that CEV-JL14 was detected in almost all tissues including the heart, liver, spleen, lung, kidney, intestine, brain, and muscle, with significantly higher viral loads in the heart, liver, lung, kidney, and intestine. These results revealed the pattern of viral load and tropism for CEV-JL14 and provided a model system for elucidating the pathogenesis of CEV-JL14 viruses.

Hydrogen bond network dynamics of heavy water resolved by alcohol hydration under an intense laser.

Despite a great deal of effort spanning for decades, it remains yet puzzling concerning how alcohol molecules functionalize the hydrogen bond (H-bond) networks of water. We employed an isotopic substitution method (using alcohol-heavy water system) to avoid spectral overlap between the alcohol hydroxyl groups and water hydrogen bonds. We showed spectrometrically that under the strong pulse laser, the low mixing ratio (VA < 20%) of alcohol can strengthen the H-bond network structure of D2O through :ÖC2H6↔ D2Ö: compression. But when VA > 20%, H-bond network of D2O will deform via the self-association between alcohol molecules. Our experiments not only reveal the H-bond kinetics of heavy water-alcohol interactions but also provide important reference for understanding the distinctive properties of H-bond in water-organic system.

Tuning the Fermi resonance of pyridine using ethanol molecules.

The Fermi resonance (FR) phenomenon is prevalent in infrared and Raman spectroscopy, and it can be observed in a variety of molecules. In particular, pyridine is a compound that has two Fermi doublets: ν1 âˆ¼ ν12 and ν1 + ν6 âˆ¼ ν8. To analyze the effect of environmental changes on the FR, this study first investigated the Raman spectra of pyridine mixed with ethanol at different concentrations. Results indicated that the FR parameters exhibited a nonlinear dependence on the pyridine concentration fractions, and changing the concentration fraction of pyridine led to different hydrogen bond strengths. Second, the interaction mechanism of pyridine-ethanol binary solutions was analyzed by two-dimensional correlation Raman spectroscopy (2DCRS). In addition, high-pressure Raman spectra of a 50% pyridine-ethanol binary solution were measured up to a pressure of 19.65 GPa by a diamond anvil cell technique, and the phase transition of the binary solution occurred at 6.35 GPa. Finally, the impact of ethanol on the FR of pyridine was determined by deducing the FR parameters at different pressures. The turning point at 6.35 GPa was consistent with the Raman frequency-pressure relationships. The results demonstrated that changes in the intensity of ν1 did not affect the FR of ν1 + ν6 âˆ¼ ν8, whereas the undisturbed frequency ν1 still played a role in the FR. When the pressure was compressed to 13.36 GPa, the disappearance of the Raman peaks (ν1 and ν1') was attributed to the tuning of the molecular symmetry by pressure during the phase transition.

Unveiling of the epidemiological patterns for caprine/ovine enterovirus infection in China.

Caprine/ovine enterovirus (CEV/OEV) infection is an emerging disease and remains largely unknown for its infection distribution, epidemic pattern, and the underlying contribution factors. Here, we report the investigation on CEV/OEV infection pattern and the underlying contribution factors by employing a sandwich ELISA kit for detection of CEV/OEV antigen. Epidemiological investigation revealed a wide range of infection rates of CEV/OEV from 19.80%-39.00% on goat/sheep farms in the major goat/sheep-raising provinces as such Henan, Shandong, Ningxia, Jilin, Inner Mongolia autonomous region, and Xinjiang autonomous region in China. Epidemic patterns and infection rates for CEV/OEV were affected by the breeds, raising mode, regions, and seasons. CEV/OEV infection rates were varied in different regions in China and significantly higher in the diarrheal herds (40.30%) than these in non-diarrheal herds (13.83%). Moreover, infection rate was higher in sheep (24.59%) than that in goats (9.84%), even dramatic difference among different breeds of goat or sheep. Out of different breeds of goat, Boer (20.13%) had the highest infection rate, followed by local breed (5.62%) and Saanen (2.61%). Among these breeds of sheep, higher infection rates were detected in local breed sheep (42.86%) and small-tailed Han sheep (35.91%) than these of Hu sheep (13.41%) and Dorper sheep (16.34%). Furthermore, raising modes were showed to contribute to the infection rate, where higher rates were detected among goats/sheep in captivity (27.10%) than these in free-range (12.27%) and semi-free range (19.24%). Additionally, CEV/OEV infection rate had obvious seasonality, while they increased from year 2015 to 2019. In summary, we investigated the CEV/OEV infection among the goat/sheep herds from different regions in China, revealed the epidemic pattern and the contribution factors to the infection, which provided the epidemiological data for future prevention and control of this emerging infection.

Molecular Analysis of Caprine Enterovirus Circulating in China during 2016-2021: Evolutionary Significance.

Here, we report the characterization of 13 novel caprine/ovine enterovirus strains isolated from different regions in China during 2016-2021. Immunoperoxidase monolayer assay showed that these viral strains shared strong cross-reaction with the previously reported caprine enterovirus CEV-JL14. Alignment analysis of the complete nucleotide sequences revealed 79.2%-87.8% and 75.0%-76.7% sequence identity of these novel caprine enterovirus strains to CEV-JL14 and TB4-OEV, respectively. Phylogenetic analyses clustered these novel strains to EV-G based on the amino acid sequences of P1 and 2C+3CD. Moreover, phylogenetic analysis of these caprine enterovirus strains identified three new EV-G types using VP1 sequences. These results demonstrate the genetic variations and the evolution of caprine enterovirus.

Unveiling of Evolution Pattern for HY12 Enterovirus Quasispecies and Pathogenicity Alteration.

Enterovirus, like the majority of RNA viruses, evolves to survive the changeable environments by a variety of strategies. Here, we showed that HY12 virus evolved to alter its characteristics and pathogenicity by employing a non-synonymous mutation. Analyses of 5'UTR, VP1 and VP2 gene sequences revealed the existence of HY12 virus in an array of mutants defined as quasispecies. The determination of diversity and complexity showed that the mutation rate and complexity of HY12 virus quasispecies increased, while the proportion of HY12 VP1 and VP2 consensus (master) sequences decreased with increasing passages. Synonymous mutation and non-synonymous mutation analysis displayed a positive selection for HY12 quasispecies evolution. A comparison of HY12 virus in different passages demonstrated that HY12 virus altered its characteristic, phenotype, and pathogenicity via non-synonymous mutation. These findings revealed the evolution pattern for HY12 virus, and the alteration of HY12 virus characteristics and pathogenicity by mutation.

Mechanism of blood-brain barrier disruption by an Escherichia coli from lambs with severe diarrhea and meningoencephalitis.

Escherichia coli (E. coli) is a common conditional pathogen that is associated with a variety of infections in humans and animals. Although there are increasing reports regarding the infection of E. coli to domestic animals and poultry, the infection of E. coli in lambs is relatively less reported, especially on meningoencephalitis. Here, we reported the isolation of an E. coli strain designated as NMGCF-19 from lambs characterized with severe diarrhea and neurological disorder, and demonstrated that NMGCF-19 as the causative agent has the ability to disrupt the blood-brain barrier (BBB) to cause the meningoencephalitis using a mouse model. Investigation on the mechanism regarding the NMGCF-19-related meningoencephalitis revealed a significant decreased expression of ZO-1 and occludin in mouse brain tissue in comparison with the control mice. Moreover, infection of NMGCF-19 increased the expression of proinflammatory cytokines TNF-α, IL-6, IL-1ß and IL-18, up-regulated HMGB1 level, and activated TLR2/TLR4/MyD88 and NLRP3 inflammasome pathways. These findings indicated that NMGCF-19 likely invades the brain tissue by disrupting the tight junction (TJ) architecture and causes the meningoencephalitis via increasing inflammatory response and activating TLR2/TLR4/MyD88 and NLRP3 inflammasome pathways.

Discovery of a virus of the species Enterovirus F in goats.

Here, we report two novel enteroviruses, designated as SD-S67 and SD-S68, isolated from a goat farm. Their complete genome sequences were determined and found to be 7455 and 7465 nucleotides in length, respectively. Molecular characterization revealed that SD-S67 is closely related to bovine enterovirus strain 261 and that SD-S68 to caprine enterovirus strain CEV-JL14. Phylogenetic analysis showed that SD-S67 clustered with members of the species Enterovirus F, and that SD-S68 clustered with enteroviruses of goats and sheep. Recombination analysis showed that SD-S67 is likely to have undergone several recombination events in the process of its evolution. To the best of our knowledge, this is the first report of an enterovirus F isolate from a goat and of a coinfection with enteroviruses of different species in the same goat herd.

Identification of three linear B cell epitopes using monoclonal antibodies against bovine enterovirus VP2 protein.

Bovine enterovirus (BEV) VP2 protein is a structural protein that plays an important role in inducing protective immunity in the host. The function of VP2 has been characterized, but there is little information on its B cell epitopes. Three monoclonal antibodies (mAbs) directed against BEV VP2 were generated and characterized from mice immunized with the recombinant VP2 protein. Three minimal linear epitopes 152FQEAFWLEDG161, 168LIYPHQ173, and 46DATSVD51 reactive to the three mAbs were identified using western blotting analysis. Three-dimensional model of the BEV-E virion and the VP2 monomer showed that epitope 152FQEAFWLEDG161 is exposed on surface of the virion and epitopes 46DATSVD51 and 168LIYPHQ173 are located inside the virion. Alignment of the amino acid sequences corresponding to the regions containing the three minimal linear epitopes in the VP2 proteins and their cross-reactivity with the three mAbs showed that epitope 168LIYPHQ173 is completely conserved in all BEV strains. Epitope 46DATSVD51 is highly conserved among BEV-E strains and partly conserved among BEV-F strains. However, epitope 152FQEAFWLEDG161 is not conserved among BEV-F strains. Using the mAbs of 3H4 and 1E10, we found that VP2 localized in the cytoplasm during viral replication and could be used to monitor the viral antigen in infected tissues using immunohistochemistry. A preliminary 3H4-epitope-based indirect ELISA allowed us to detect anti-BEV-strain-HY12 antibodies in mice. This study indicates that the three mAbs could be useful tools for investigating the structure and function of the viral VP2 protein and the development of serological diagnostic techniques for BEV infection.

Disruption of blood-brain barrier by an Escherichia coli isolated from canine septicemia and meningoencephalitis.

Escherichia coli (E. coli) is one of the common pathogenic bacteria in veterinary clinical infection. As an opportunistic microorganism, E. coli normally does not cause diseases. However, it causes infections under certain circumstance to domesticated animal and poultry, resulting in severe diarrhea, septicemia, and respiratory infections. Although there are increasing reports regarding the infections of E. coli to domestic animals and poultry, the infection of E. coli in dogs is relatively less reported, especially on septicemia and meningoencephalitis. Here, we reported the isolation and identification of an E. coli isolate named CEC-GZL17 from dogs characterized by septicemia and sudden death, and found that CEC-GZL17 is able to cause meningoencephalitis. Exploration on the potential mechanism underlying meningoencephalitis demonstrated that CEC-GZL17 infection significantly increases TNF-α expression and inhibits ZO-1 and occludin expressions in brain tissue, indicating that the E coli likely use the mechanism to penetrate the blood-brain barrier via disrupting tight junction architecture, thus leading to the invasion to brain tissue.

Rescue and characterization of a recombinant HY12 bovine enterovirus carrying a foreign HA epitope in the 3A nonstructural protein.

Full-length infectious cDNA clones for recombinant HY12 bovine enteroviruses designated as rHY12-3A-2-HA, rHY12-3A-3-HA, and rHY12-3A-9-HA were constructed by the insertion of an epitope from influenza virus hemagglutinin (HA) at the N-terminus of the HY12-encoded 3A protein at amino acid positions 2, 3, and 9. The recombinant HY12 viruses expressing the HA epitope were rescued and characterized using immunoperoxidase monolayer assay, western blotting, and electron microscopy. The three rescued recombinant marker viruses showed similar characteristics, such as TCID50 titer, plaque size, and growth properties, to those of parental rHY12 virus. Comparative analysis of the nucleotide sequences demonstrated the three recombinant marker viruses remained stable for 15 passages with no genetic changes. The recombinant viruses remained viable in various permissive cell lines, including BHK-21, Vero, and PK15 cells, suggesting that the insertion of the HA epitope tag had no effect on virus infectivity. Mice infected with the recombinant marker viruses and the parental virus produced anti-HY12-virus antibodies, while the recombinant marker viruses also produced anti-HA-epitope-tag antibodies. Taken together, these results demonstrate that HY12 viruses containing genetic markers may be useful tools for future investigations of the mechanisms of viral pathogenesis and virus replication, as well as for vaccine development.

Construction and evaluation of HA-epitope-tag introduction onto the VP1 structural protein of a novel HY12 enterovirus.

In this study, we investigated the feasibility of using enterovirus HY12 as a vector to express an exogenous hemagglutinin (HA)-epitope tag onto the HY12-encoded VP1 protein via a reverse genetic system. Characteristics of recombinant (r) HY12-VP1-HA marker virus were determined by immunoperoxidase monolayer assay, western blot, electron microscopy, and serum-neutralisation assay. Sequence analysis demonstrated that the marker virus stably maintained the HA-epitope-tag in MDBK cells, with no changes in viral morphological features observed relative to those of the parental rHY12 virus. Furthermore, detection by immunofluorescence assay revealed the expression of HA-epitope tag and VP2 protein, which distinguish the marker virus from parental rHY12 virus. In addition, neonatal mice infected with the recombinant marker virus showed various microscopic pathological lesions and generated anti-HY12 virus and -HA-epitope-tag antibodies. These results indicated that the recombinant marker virus represented a valuable platform to promote the development of novel genetic vaccines.

Corrosion Behaviors of Q345R Steel at the Initial Stage in an Oxygen-Containing Aqueous Environment: Experiment and Modeling.

The ingress of oxygen into pressure vessels used in oil & gas production and transportation could easily result in serious corrosion. In this work, the corrosion behaviors of Q345R steel at the initial stage in 1 wt.% NaCl solution were investigated using electrochemical techniques. The effects of oxygen concentration, temperature and pH on corrosion behaviors were discussed. Simultaneously, a numerical model based on the mixed potential theory was proposed. The results show that the proposed model accords well with the experimental data in the pH range from 9.0 to 5.0. In this pH range, the oxygen reduction reaction, H⁺ reduction, water reduction, and iron oxidation can be quantitatively analyzed using this model. However, this model shows a disagreement with the experimental data at lower pH. This can be attributed to the fact that actual area of reaction on the electrode is much smaller than the preset area due to the block effect resulted from hydrogen bubbles adsorbed on the electrode surface.

A fatal suppurative pneumonia in piglets caused by a pathogenic coagulase-positive strain of Staphylococcus hyicus.

Staphylococcus hyicus is one of the opportunistic pathogens that cause infections to animals. Early studies have demonstrated that S. hyicus is the causative agents of exudative epidermitis in pigs, arthritis in horses and chicken, mastitis in cow, and bacteremia, sepsis and multiple organ failure in humans. Here, we report the isolation and identification of a representative S. hyicus isolate, named JLHN15, from a pig farm with a disease characterized by bacteremia, suppurative pneumonia and fibrinous pericarditis. Our results indicate that JLHN15 is a pathogenic coagulase-positive Staphylococcus. To the best knowledge, this is the first report of S. hyicus causing an infection characterized by suppurative pneumonia and sepsis.