Search

This paper describes the performance of the Regional Atmospheric Modeling System (RAMS) in simulating a winter drought event, based on two different forcing datasets. We ran EC (ERA-Interim reanalysis data as initial and lateral boundary forcing conditions) and FNL (NCEP-FNL reanalysis data) simulations for the 2008/2009 winter drought event to quantify the impact of any uncertainty in the different initial and lateral boundary forcing data on regional model outputs. The response of the winter mean atmospheric states to the variations in the initial and lateral boundary conditions was investigated on the basis of these simulation results. The spatio-temporal features of precipitation from the EC and FNL runs closely resembled those measured from the Global Summary Of the Day (GSOD) observations, although the EC run data outperformed the FNL run data in both their spatial distribution patterns and precipitation values. The water vapor flux values explain how the differences in the precipitation values between the EC and the FNL runs were generated, whereas temperature values were not sensitive to any changes in forcing data. The model results from these runs also slightly overestimated temperature on both spatial and temporal scales. For the tropospheric atmospheric data recorded at the Fuyang Meteorological Station in Anhui Province, neither the time series nor the statistical analyses showed any evidence of superiority between the two different driver datasets compared with radiosonde data. However, on closer inspection, the influence of different initial and lateral boundary conditions on modeling the tropospheric atmospheric data appeared to be evident. [ABSTRACT FROM AUTHOR]

A field canine parvovirus (CPV) strain, CPV-SH14, was previously isolated from an outbreak of severe gastroenteritis in Shanghai in 2014. The complete genome of CPV-SH14 was determined by using PCR with modified primers. When compared to other CPV-2 strains, several insertions, deletions, and point mutations were identified in the 5’ and 3’ UTR, with key amino acid (aa) mutations (K19R, E572K in NS1 and F267Y, Y324I and T440A in VP2) also being observed in the coding regions of CPV-SH14. These results indicated that significant and unique genetic variations have occurred at key sites or residues in the genome of CPV-SH14, suggesting the presence of a novel genetic variant of new CPV-2a. Phylogenetic analysis of the VP2 gene revealed that CPV-SH14 may have the potential to spread worldwide. In conclusion, CPV-SH14 may be a novel genetic variant of new CPV-2a, potentially with a selective advantage over other strains. [ABSTRACT FROM AUTHOR]

Infectious bronchitis (IB) and Newcastle disease (ND) are two major infectious diseases that are a threat to the domestic poultry industry. In this study, we successfully generated a recombinant LaSota candidate vaccine strain, rNDV-IBV-T/B, which expresses a short, synthetic, previously identified IBV S1 multi-epitope cassette using the reverse genetic system. The recombinant virus was propagated in nine-day-old embryonated chicken eggs for 20 passages and genetic stability was confirmed by whole genome DNA sequencing. The recombinant virus had a hemagglutination (HA) titer of 210, mean death time (MDT) of 118 hours, and intracerebral pathogenicity index (ICPI) of 0.05. None of these were significantly different from the parental Newcastle disease virus (NDV) LaSota strain (p > 0.05). Vaccination of white leghorn chickens at one day of age with 106 EID50 rNDV-IBV-T/B provided 90% protection against virulent IBV M41 challenge at three weeks of age, which was significantly higher than the protection of the control group vaccinated with phosphate-buffered saline (PBS) (p < 0.05). The ciliostasis scores of rNDV-IBV-T/B-vaccinated and LaSota-vaccinated groups were 4.2 and 37.6, respectively, which indicated that rNDV-IBV-T/B vaccination reduced the pathogenicity of IBV toward the trachea. Furthermore, real-time RT-PCR assay showed that the rNDV-IBV-T/B vaccination resulted in low levels of viral load (647.80 ± 49.65 RNA copies) in the trachea four days post-challenge, which is significantly lower than groups vaccinated with PBS (8591.25 ± 311.10 RNA copies) or LaSota (7742.60 ± 298.50 RNA copies) (p < 0.05). Meanwhile, the same dose of rNDV-IBV-T/B vaccination provided complete protection against velogenic NDV F48E9 challenge. These results demonstrate that the rNDV-IBV-T/B strain is a promising vaccine candidate to control both IB and ND simultaneously. Furthermore, epitope-based live vector vaccines provide an alternative strategy for the development of cost-effective and, broadly, cross-protective vaccines. [ABSTRACT FROM AUTHOR]

In aiming for better access to climate change information and for providing climate service, it is important to obtain reliable high-resolution temperature simulations. Systematic comparisons are still deficient between statistical and dynamic downscaling techniques because of their inherent unavoidable uncertainties. In this paper, 20 global climate models (GCMs) and one regional climate model [Providing Regional Climates to Impact Studies (PRECIS)] are employed to evaluate their capabilities in reproducing average trends of mean temperature (Tm), maximum temperature (Tmax), minimum temperature (Tmin), diurnal temperature range (DTR), and extreme events represented by frost days (FD) and heat-wave days (HD) across China. It is shown generally that bias of temperatures from GCMs relative to observations is over ±1°C across more than one-half of mainland China. PRECIS demonstrates better representation of temperatures (except for HD) relative to GCMs. There is relatively better performance in Huanghuai, Jianghuai, Jianghan, south Yangzi River, and South China, whereas estimation is not as good in Xinjiang, the eastern part of northwest China, and the Tibetan Plateau. Bias-correction spatial disaggregation is used to downscale GCMs outputs, and bias correction is applied for PRECIS outputs, which demonstrate better improvement to a bias within ±0.2°C for Tm, Tmax, Tmin, and DTR and ±2 days for FD and HD. Furthermore, such improvement is also verified by the evidence of increased spatial correlation coefficient and symmetrical uncertainty, decreased root-mean-square error, and lower standard deviation for reproductions. It is seen from comprehensive ranking metrics that different downscaled models show the most improvement across different climatic regions, implying that optional ensembles of models should be adopted to provide sufficient high-quality climate information. [ABSTRACT FROM AUTHOR]

Background: Felid herpesvirus 1 (FHV-1) is a major pathogenic agent of upper respiratory tract infections and eye damage in felines worldwide. Current FHV-1 vaccines offer limited protection of short duration, and therefore, do not reduce the development of clinical signs or the latency of FHV-1. Methods: To address these shortcomings, we constructed FHV ∆gIgE-eGFP, FHV ∆TK mCherry, and FHV ∆gIgE/TK eGFP-mCherry deletion mutants (ΔgI/gE, ΔTK, and ΔgIgE/TK, respectively) using the clustered regularly interspaced palindromic repeats (CRISPR)/CRISP-associated protein 9 (Cas9) system (CRISPR/Cas9), which showed safety and immunogenicity in vitro. We evaluated the safety and efficacy of the deletion mutants administered with intranasal (IN) and IN + subcutaneous (SC) vaccination protocols. Cats in the vaccination group were vaccinated twice at a 4-week interval, and all cats were challenged with infection 3 weeks after the last vaccination. The cats were assessed for clinical signs, nasal shedding, and virus-neutralizing antibodies (VN), and with postmortem histological testing. Results: Vaccination with the gI/gE-deleted and gI/gE/TK-deleted mutants was safe and resulted in significantly lower clinical disease scores, fewer pathological changes, and less nasal virus shedding after infection. All three mutants induced virus-neutralizing antibodies after immunization. Conclusions: In conclusion, this study demonstrates the advantages of FHV-1 deletion mutants in preventing FHV-1 infection in cats. [ABSTRACT FROM AUTHOR]

Virulent systemic feline calicivirus (VS-FCV) is a newly emerging FCV variant that is associated with a severe acute multisystem disease in cats that is characterized by jaundice, oedema, and high mortality (approximately 70%). VS-FCV has spread throughout the world, but there are no effective vaccines or therapeutic options to combat infection. VS-FCV may therefore pose a serious threat to the health of felines. The genomic characteristics and functions of VS-FCV are still poorly understood, and the reason for its increased pathogenicity is unknown. Reverse genetics systems are powerful tools for studying the molecular biology of RNA viruses, but a reverse genetics system for VS-FCV has not yet been reported. In this study, we developed a plasmid-based reverse genetics system for VS-FCV in which infectious progeny virus is produced in plasmid-transfected CRFK cells. Using this system, we found that the 3' untranslated region (UTR) and poly(A) tail are important for maintaining the infection and replication capacity of VS-FCV and that shortening of the poly(A) tail to less than 28 bases eliminated the ability to rescue infectious progeny virus. Whether these observations are unique to VS-FCV or represent more-general features of FCV remains to be determined. In conclusion, we successfully established a rapid and efficient VS-FCV reverse genetics system, which provides a good platform for future research on the gene functions and pathogenesis of VS-FCV. The effects of the deletion of 3' UTR and poly(A) tail on VS-FCV infectivity and replication also provided new information about the pathogenesis of VS-FCV. [ABSTRACT FROM AUTHOR]

Background: Peste des petits ruminants (PPR) disease is a cross-species infectious disease that severely affects small ruminants and causes great losses to livestock industries in various countries. Distinguishing vaccine-immunized animals from naturally infected animals is an important prerequisite for the eradication of PPR. At present PPRV are classified into lineages I through IV, and only one vaccination strain, Nigeria/75/1, belongs to lineage II, but all of the epidemic strains in China at present are from lineage IV. Results: To achieve this goal, we developed an SYBR Green I real-time qRT-PCR method for rapid detection and identification of PPRV lineages II and IV by analyzing different melting curve analyses. The negative amplification of other commonly circulating viruses such as orf virus, goat poxvirus, and foot-and-mouth disease virus demonstrated that primers targeting the L gene of PPRV were extremely specific. The sensitivity of the assay was assessed based on plasmid DNA and the detection limit achieved was 100 copies of PPRV lineages II and IV. Conclusion: Since the method has high sensitivity, specificity, and reproducibility, it will be effectively differentiated PPRV lineages II from PPRV lineages IV in PPRV infected animals. [ABSTRACT FROM AUTHOR]

Abstract Mitochondrial antiviral-signaling protein (MAVS) is an essential adaptor protein in retinoic acid-inducible gene I (RIG-I)-mediated antiviral innate immunity in mammals. In this study, the goose MAVS gene (goMAVS) was identified. The 2019 bp-long goMAVS exhibits 96.2% amino acid similarity compared to the predicted goMAVS. Quantitative real-time polymerase chain reactions showed that goMAVS mRNA was widely expressed in different tissues. The overexpression of goMAVS in goose embryo fibroblast cells up-regulated the mRNA levels of goose interferon-stimulated genes. We concluded that MAVS mediates the activation of type I interferon (IFN) pathway in a species-specific manner. We further demonstrated that a CARD-like domain, transmembrane domain and two previously unidentified domains of goMAVS were essential for the activation of type I IFN pathway. GoMAVS inhibited Newcastle disease virus replication by activating type I IFN pathways, especially at the early stages of infection. Finally, the interaction between goMAVS and goose RIG-I was confirmed. The CARD domain of goMAVS plays a vital role in the interaction. Together, we identified goMAVS as a goRIG-I interactive protein and concluded that goMAVS is involved in the activation of type I IFN pathways in goose cells. Highlights • Goose MAVS (goMAVS) gene was identified for the first time. • GoMAVS plays an antiviral role by activating type I interferon (IFN) pathway in a species-specific manner. • The CARD-like domain, transmembrane domain of goMAVS were essential for goMAVS function. • The CARD-like domain of goMAVS interacts with goose RIG-I. [ABSTRACT FROM AUTHOR]

Knowledge about the regionalization of the planetary boundary layer (PBL) height (PBLH) remains unclear for such a huge country like China, largely to the limited observations. By using K-means clustering of summer radiosonde-derived PBLHs, combined with land surface related factors, we classified China into six regions, including Northeast China, North China, South China, Inner Mongolia, Tibet Plateau, and Northwest China. Overall, the PBLH in China generally reaches the minimum at 0800 Beijing time (BJT), as opposed to the peak at 1400 BJT when the convective PBL is well developed. Concerning the spatial distribution, the PBLH at 14 BJT exhibits a "northwest high-southeast low" pattern with the highest (2259 m) occurring in Inner Mongolia. Furthermore, the performances of several commonly used reanalysis products (ERA5, FNL, and CERA) are comprehensively compared against PBLH. Among all the three reanalysis datasets, The PBLHs from ERA5 present the strongest relationship with those from radiosounding observations across China, suggesting its robustness and suitability in estimating PBLHs. In terms of the performance of the reanalysis dataset in characterizing three PBL regimes such as convective, neutral and stable PBL, the PBLHs from ERA5 show significant positive correlations (R > 0.7) with the observed counterparts for both convective and neutral PBL regimes in most areas. However, all three reanalysis datasets tend to overestimate the PBLH under stable SBL. Our findings indicate that PBLH biases in reanalysis products are partly attributed to the misrepresentation of PBL thermal regimes, which is probably owing to the unsatisfactory simulations of cloud effects in weather models. Overall, this study presents the first regionalization of high-resolution radiosonde-derived PBLHs in China and quantitatively evaluates three popular reanalysis products, which have great implications for better application of datasets and improve the model parameterization of PBL. • Six regions were classified based on clustering of the PBLHs over China. • All the reanalysis products severely overestimate early-afternoon SBL heights. • ERA5 is robust in estimating PBLHs over China compared to FNL and CERA. [ABSTRACT FROM AUTHOR]

• A novel surface-anchored photocatalytic nanofiltration membrane was fabricated. • A continuous flow-through process under visible light irradiation was involved. • It removes antibiotics in sewage effluent indiscriminately to more than 99%. • Good self-cleaning and biotoxicity reduction capacities were also fulfilled. • The usability of photocatalytic nanofiltration membrane is greatly improved. Bifunctional photocatalytic nanofiltration (PNF) membrane is increasingly concerned in practical micro-polluted water purification, but there are still several bottlenecks that inhibit its practicality. In this context, the feasibility of a novel metal-free and visible light-responsive surface-anchored PNF membrane for simultaneously removing target antibiotics in real sewage effluent in a continuous dynamic process was explored. The results showed that the optimal PNF-4 membrane was expectedly consisted of an inside tight sub-nanopore structured separation layer and an outside thinner, smoother, super hydrophilic mesoporous degradation layer, respectively. Consequently, the activated PNF-4 membrane could synergistically reduce trimethoprim and sulfamethoxazole concentrations to below two orders of magnitude, accompanying with almost constant high water permeability, suggesting that the hydrophilic modification of the mesoporous degradation layer basically offsets its inherent hydraulic resistance. Also, after repeating the fouling-physical rinsing process three times lasted for 78 h, only sporadic adherent contaminants remained onto the top surface, together with the minimal total and irreversible fouling ratios (as low as 7.2% and 1.2%, respectively), strongly demonstrated that PNF-4 membrane displayed good self-cleaning performance. Undoubtedly, this will significantly reduce its potential cleaning frequency and maintenance cost in long-term operation. Meanwhile, the acute and chronic biotoxicities of its permeate to Virbrio qinghaiensis sp. -67 were also reduced sharply to 2.22% and 0.45%, respectively. All of these evidences suggest that the dual functions of PNF-4 membrane are synergetic in an uninterrupted permeating process. It will provide useful insights for continuously enhancing the practicality and effectiveness of PNF membrane in actual micro-polluted water purification scenarios. [Display omitted] [ABSTRACT FROM AUTHOR]

Rabbit hemorrhagic disease virus (RHDV) is a major member of the Caliciviridae. which is fatal to wild and domestic European rabbit. Because RHDV does not reproduce stably in vitro , molecular studies on this pathogen have been limited. Feline calicivirus (FCV), also a member of the Caliciviridae , reproduces well in vitro and is a good viral vector. As these viruses share similar genomic structures, we hypothesized that a chimeric infectious clone could be constructed by replacing the corresponding regions of the FCV genome with the structural proteins VP60 and VP10 and the 3′ non-translated region of the RHDV genome. Transfection of the infectious clone into RK13 cells made it possible to rescue the chimeric virus, named pseudoRHDV, which reproduced in an RK13 cell line with high titer. An infectious pseudoRHDV was produced, which proliferated in RK13 cells to at least 15 generations. PseudoRHDV caused significant cytopathic changes in the RK13 cells, with a viral titer was 9.74 log 10 TCID 50 / mL. The pseudoRHDV constructed in this study will be helpful for investigating the molecular biology of RHDV, especially its interaction with the host. The model can also be used to explore some common laws between FCV and RHDV. • PseudoRHDV constructed with feline calicivirus genome as vector can efficiently reproduce in multiple cell lines. • PseudoRHDV as a model can be used to explore some common laws between FCV and RHDV, even for calicivirus. • The virus titer of pseudoRHDV is higher than that of its parent virus. [ABSTRACT FROM AUTHOR]

Land surface temperature (LST) is a key parameter in the physics of land surface processes. Currently, the technique most commonly used to obtain LST is thermal infrared (TIR) remote sensing (RS). However, this technique is affected by clouds and cannot obtain complete spatiotemporal LST images. To solve this problem, RS and weather research and forecasting (WRF) coupled model (RS-WRF coupled model) was developed to produce cloud-free MODIS-like LST data. (i) The WRF model is used to simulate the cloud-free LST with a 1 km resolution. (ii) The optimal machine learning model is utilized to fit the simulated LSTs and produce cloud-free MODIS-like LSTs. (iii) Combined with a median filtering algorithm, the salt and pepper noise in the fitted image is optimized. Taking Beijing as a test site. Under relatively little cloud cover and greater cloud contamination, the root mean square error of the LST constructed by the RS-WRF coupled model is approximately 1.2 and 1.8 K, respectively. The correlation coefficients under both conditions exceed 0.9. Overall, the RS-WRF coupled model can provide cloud-free time series MODIS-like LST images in areas with frequent cloud cover, thereby compensating for the disadvantage that satellite TIR images contaminated by clouds cannot obtain complete LST estimates. • A new method is developed to construct cloud-free MODIS-like LSTs. • The optimal WRF scheme is used to simulate the LST. • The optimal machine learning model and median filter are used to improve the accuracy of LST construction. • The errors in the LSTs constructed under little/considerable cloud contamination are around 1.2 K and 1.8 K, respectively. [ABSTRACT FROM AUTHOR]

The APOBEC proteins play significant roles in the innate and adaptive immune system, probably due to their deaminase activities. Because APOBEC1 (A1) and APOBEC3 (A3) are absent in the chicken genome, we were interested in determining whether chicken APOBEC4 (A4) possessed more complex functions than its mammalian homologs. In this study, chicken A4 (chA4) mRNA was identified and cloned for the first time. Based on bioinformatics analyses, the conserved zinc-coordinating motif (HXE ... PC(X) 2–6 C) was identified on the surface of chA4 and contained highly conserved His97, Glu99, Pro130, Cys131 and Cys138 active sites. The highest expression levels of constitutive chA4 were detected in primary lymphocytes and bursa of Fabricius. Newcastle Disease (ND) is one of the most serious infectious diseases in birds, causing major economic losses to the poultry industry. In vitro , Newcastle Disease Virus (NDV) early infection induced significant increases in chA4 expression in the chicken B cell line, DT40, the macrophage cell line, HD11 and the CD4+ T cell line, MSB-1, but not the fibroblast cell line, DF-1. In vivo , the expression levels of chA4 were up-regulated in several tissues from NDV-infected chickens, especially the thymus, testicles, duodenum and kidney. The high level expression of exogenous chA4 displayed inhibitory effects on NDV and reduced viral RNA in infected cells. Taken together, these data demonstrate that chA4 is involved in the chicken immune system and may play important roles in host anti-viral responses. • First time isolation and identification of chicken APOBEC4. • chA4 contains typical APOBEC motifs and probably has catalytic activity. • The highest expression levels of constitutive chA4 were detected in primary lymphocytes and bursa of Fabricius. • NDV infection induces significant increases in chA4 expression in immune cells and in the tissues of infected chickens. • chA4 expression induces inhibitory effects on NDV. [ABSTRACT FROM AUTHOR]

Bifunctional photocatalytic nanofiltration (PNF) membrane has become a reliable frontier technique for removing refractory organic micropollutants. However, the active mitigated fouling mechanism from the microscopic perspective during its long-term operation of purifying real micro-polluted water is rarely studied. Herein, with an integrated use of QSense Explorer and confocal laser scanning microscope techniques, self-cleaning foulant attachment on an activated and customized near-infrared responsive polymeric PNF (termed as nPNF) membrane with good service performance for continuous dynamic removing antibiotics in sewage effluent environment was firstly elucidated. Time-dependent changes in dissipation oscillation frequency, sensed mass and the visualized foulant spatial distribution all indicated that there were only sporadic foulant attachment, an extremely low fouling layer thickness and irreversible fouling rate on/of the activated nPNF membrane top surface, thereby endowing it with excellent self-cleaning characteristic. This is probably because the reactive oxygen species (mainly •O 2 - and •OH) concurrently destroys the integrity of fouling layer and its internal adhesion structure, transforming part of the irreversible fouling on nPNF membrane surface into reversible one that is easy to wash off. These new horizons provided useful insight on the fate of selected antibiotics in the to-be-removed stage and self-cleaning foulant attachment of PNF membrane., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Duck hepatitis B virus (DHBV) infection model was frequently used as the experimental model for human hepatitis B virus (HBV) research. In order to decipher the genetic characteristics of DHBVs from Anhui province of China, 120 duck liver tissue samples were collected and subjected to PCR screening, and 28 samples were detected as DHBV positive. Subsequently, five DHBV-positive samples were selected for genome-wide amplification and a comprehensive analysis. Comparative analysis of complete genome sequences using the MegAlign program showed that five strains of DHBVs shared 94.5-96.3% with each other and 93.2-98.7% with other reference strains in GenBank. The phylogenetic analysis showed that all five DHBV strains belonged to the evolutionary branch of "Chinese DHBV" isolates or DHBV-2. Importantly, three potential intra-genotypic recombination events, between strains AAU-6 and Guilin, strains AAU-1 and GD3, and strains AAU-6 and AAU-1, were respectively found using the RDP and SimPlot softwares and considered the first report in avihepadnaviruses. These results not only improve our understanding for molecular prevalence status of DHBV among ducks, but also provide a reference for recombination mechanism of HBV., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Elucidating virus-cell interactions is fundamental to understanding viral replication and identifying targets for therapeutic control of viral infection. The extracellular signal-regulated kinase (ERK) pathway has been shown to regulate pathogenesis during many viral infections, but its role during coronavirus infection is undetermined. Infectious bronchitis virus is the representative strain of Gammacoronavirus, which causes acute and highly contagious diseases in the poultry farm. In this study, we investigated the role of ERK1/2 signaling pathway in IBV infection. We found that IBV infection activated ERK1/2 signaling and the up-regulation of phosphatase DUSP6 formed a negative regulation loop. Pharmacological inhibition of MEK1/2-ERK1/2 signaling suppressed the expression of DUSP6, promoted cell death, and restricted virus replication. In contrast, suppression of DUSP6 by chemical inhibitor or siRNA increased the phosphorylation of ERK1/2, protected cells from apoptosis, and facilitated IBV replication. Overexpression of DUSP6 decreased the level of phospho-ERK1/2, promoted apoptosis, while dominant negative mutant DUSP6-DN lost the regulation function on ERK1/2 signaling and apoptosis. In conclusion, these data suggest that MEK-ERK1/2 signaling pathway facilitates IBV infection, probably by promoting cell survival; meanwhile, induction of DUSP6 forms a negative regulation loop to restrict ERK1/2 signaling, correlated with increased apoptosis and reduced viral load. Consequently, components of the ERK pathway, such as MEK1/2 and DUSP6, represent excellent targets for the development of antiviral drugs.

To construct a eukaryotic expression plasmid containing the luciferase reporter gene (Fluc) to quickly detect apoptosis. Four amino acids, Asp-Glu-Val-Asp (DEVD), the recognize motif of Caspase-3, were introduced into the middle of the Fluc-C and N fragment. Meanwhile, four amino acids, Asp-Glu-Val-Gly (DEVG), were selected as a negative control. Subsequently, the recombinant gene was cloned into the N and C terminal end of the split intein, and named as pFluc-DEVD and pFluc-DEVG. Then the plasmids were transfected into cells and renilla luciferase was co-transfected in each sample as an internal control for transfection efficiency. Then the apoptosis level was detected by the double luciferase reporter gene and the Western blotting analysis. The results showed that when apoptosis occurred, the content of firefly luciferase expressed in the pFluc-DEVD plasmid transfected group was about 3 times higher than pFluc-DEVG plasmid transfected group. Furthermore, Western blotting detection indicated that the Fluc level was significantly increased in pFluc-DEVD transfected group when pre-treated by apoptosis stimulants. The activation degree of Caspase-3 was closely related to the expression of Fluc, and had a significant statistical difference. These results confirmed that firefly luciferase protein expressed by pFluc-DEVD plasmid can be cleaved by the intracellular Caspase-3 enzyme, and this plasmid can accurately reflect the cell apoptosis level, which provides a useful method for quantitative detection of apoptosis.

Mammalian cells respond to various environmental stressors to form stress granules (SGs) by arresting cytoplasmic mRNA, protein translation element, and RNA binding proteins. Virus-induced SGs function in different ways, depending on the species of virus; however, the mechanism of SG regulation of virus replication is not well understood. In this study, Newcastle disease virus (NDV) triggered stable formation of bona fide SGs on HeLa cells through activating the protein kinase R (PKR)/eIF2α pathway. NDV-induced SGs contained classic SG markers T-cell internal antigen (TIA)-1, Ras GTPase-activating protein-binding protein (G3BP)-1, eukaryotic initiation factors, and small ribosomal subunit, which could be disassembled in the presence of cycloheximide. Treatment with nocodazole, a microtubule disruption drug, led to the formation of relatively small and circular granules, indicating that NDV infection induces canonical SGs. Furthermore, the role of SGs on NDV replication was investigated by knockdown of TIA-1 and TIA-1-related (TIAR) protein, the 2 critical components involved in SG formation from the HeLa cells, followed by NDV infection. Results showed that depletion of TIA-1 or TIAR inhibited viral protein synthesis, reduced extracellular virus yields, but increased global protein translation. FISH revealed that NDV-induced SGs contained predominantly cellular mRNA rather than viral mRNA. Deletion of TIA-1 or TIAR reduced NP mRNA levels in polysomes. These results demonstrate that NDV triggers stable formation of bona fide SGs, which benefit viral protein translation and virus replication by arresting cellular mRNA.-Sun, Y., Dong, L., Yu, S., Wang, X., Zheng, H., Zhang, P., Meng, C., Zhan, Y., Tan, L., Song, C., Qiu, X., Wang, G., Liao, Y., Ding, C. Newcastle disease virus induces stable formation of bona fide stress granules to facilitate viral replication through manipulating host protein translation., (© FASEB.)

Unlabelled: It has been reported that lentogenic Newcastle disease virus (NDV) isolates have the potential to become velogenic after their transmission and circulation in chickens, but the underlying mechanism is unclear. In this study, a highly velogenic NDV variant, JS10-A10, was generated from the duck-origin lentogenic isolate JS10 through 10 consecutive passages in chicken air sacs. The velogenic properties of this selected variant were determined using mean death time (MDT) assays, intracerebral pathogenicity index (ICPI), the intravenous pathogenicity index (IVPI), histopathology, and the analysis of host tissue tropism. In contrast, JS10 remained lentogenic after 20 serial passages in chicken eggs (JS10-E20). The JS10, JS10-A10, and JS10-E20 genomes were sequenced and found to be nearly identical, suggesting that both JS10-A10 and JS10-E20 were directly generated from JS10. To investigate the mechanism for virulence enhancement, the partial genome covering the F0 cleavage site of JS10 and its variants were analyzed using ultradeep pyrosequencing (UDPS) and the proportions of virulence-related genomes in the quasispecies were calculated. Velogenic NDV genomes accumulated as a function of JS10 passaging through chicken air sacs. Our data suggest that lentogenic NDV strains circulating among poultry might be a risk factor to future potential velogenic NDV outbreaks in chickens., Importance: An avirulent isolate, JS10, was passaged through chicken air sacs and embryos, and the pathogenicity of the variants was assessed. A virulent variant, JS10-A10, was generated from consecutive passage in air sacs. We developed a deep-sequencing approach to detect low-frequency viral variants across the NDV genome. We observed that virulence enhancement of JS10 was due to the selective accumulation of velogenic quasispecies and the concomitant disappearance of lentogenic quasispecies. Our results suggest that because it is difficult to avoid contact between natural waterfowl reservoirs and sensitive poultry operations, circulating lentogenic NDV strains may represent a potential reservoir for emergent velogenic NDV strains that could cause outbreaks in chickens., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Rabbit hemorrhagic disease (RHD) is contagious and highly lethal. Commercial vaccines against RHD are produced from the livers of experimentally infected rabbits. Although several groups have reported that recombinant subunit vaccines against rabbit hemorrhagic disease virus (RHDV) are promising, application of the vaccines has been restricted due to high production costs or low yield. In the present study, we performed codon optimization of the capsid gene to increase the number of preference codons and eliminate rare codons in Spodoptera frugiperda 9 (Sf9) cells. The capsid gene was then subcloned into the pFastBac plasmid, and the recombinant baculoviruses were identified with a plaque assay. As expected, expression of the optimized capsid protein was markedly increased in the Sf9 cells, and the recombinant capsid proteins self-assembled into virus-like particles (VLPs) that were released into the cell supernatant. Rabbits inoculated with the supernatant and the purified VLPs were protected against RHDV challenge. A rapid, specific antibody response against RHDV was detected by an ELISA in all of the experimental groups. In conclusion, this strategy of producing a recombinant subunit vaccine antigen can be used to develop a low-cost, insect cell-derived recombinant subunit vaccine against RHDV.