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Raman spectroscopy is a powerful technique that enables fingerprinting of materials, molecules, and chemical environments by probing vibrational resonances. In many applications, the desired Raman signals are masked by fluorescence, either from the molecular system being studied, or from adjacent metallic nanostructures. Here, it is shown that wavelength‐modulated Raman spectroscopy provides a powerful way to significantly reduce the strength of the fluorescence background, thereby allowing the desired Raman signals to be clearly recorded. This approach is made use of to explore Raman scattering in the context of vibrational strong coupling, an area that has thus far been problematic to visualise. Specifically, strong coupling between the vibrational modes in a polymer and two types of confined light field, the fundamental mode of a metal‐clad microcavity, and the surface‐plasmon modes of an adjacent thin metal film are looked at. While clear advantages in using the wavelength‐modulated Raman approach are found, these results on strong coupling are inconclusive, and highlight the need for more work in this exciting topic area. [ABSTRACT FROM AUTHOR]

In the construction of intelligent transportation systems, short-term traffic flow prediction is of great significance for the advancement of traffic network management. But due to the presence of many complex factors in both spatial and temporal domains, it remains a complex and challenging task. Existing literature usually employs the convolutional neural network (CNN)-based methods in capturing spatiotemporal correlations. These CNN-based methods often use a single-channel data structure to represent different periodic patterns, which makes the model susceptible to over-parameterization when capturing periodic dependencies and prone to information loss after convolution. To overcome these limitations, this paper presents a hybrid deep learning method for short-term traffic flow prediction. In this method, a video-shaped multi-channel data structure is designed to represent different periodic patterns more efficiently. Next, a new mixed-pointwise convolution is introduced for capturing periodic dependencies without the negative impacts mentioned above. Lastly, an improved channel attention mechanism is proposed to learn channel inter-dependencies with controllable parameter usage. The proposed method is lightweight, yet highly effective. Compared to the state-of-the-art baseline method, it reduces the root mean squared error by up to 6.7% on the PeMSD4 dataset and 13.3% on the PeMSD7 dataset, while also achieving substantial improvement in two additional metrics, exhibiting strong robustness and great scalability across various settings. [ABSTRACT FROM AUTHOR]

Medication recommendation aims to provide clinicians with safe medicine combinations for the treatment of patients. Existing medication recommendation models are built based on the temporal structured code data of electronic health records (EHRs) in the medical database; nevertheless, unstructured data in EHRs, such as textual data containing rich information, are underexploited. To fill the gap, a novel multimodal interactive fusion network (MIFNet) is proposed for medication recommendation, which integrates both structured code information and unstructured text information in EHRs. Our model first extracts a series of informative feature representations to encode comprehensive patient health history and control potential drug–drug interactions (DDI), including medical code, clinical notes, and the DDI knowledge graph. Next, a novel cross-modal interaction extraction block is proposed to capture the intricate interaction information between the two modalities. Finally, a multimodal fusion block is adopted to fuse the constructed features and generate a medication combination list. Experiments are conducted on the public MIMIC-III dataset, and the results demonstrate that the proposed model outperforms the state-of-the-art medication recommendation methods on main evaluation metrics. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) remains a leading cause of death worldwide. Lipid rich, phenotypically antibiotic tolerant, bacteria are more resistant to antibiotics and may be responsible for relapse and the need for long-term TB treatment. We present a microfluidic system that acoustically traps live mycobacteria, M. smegmatis, a model organism for M. tuberculosis. We then perform optical analysis in the form of wavelength modulated Raman spectroscopy (WMRS) on the trapped M. smegmatis for up to eight hours, and also in the presence of isoniazid (INH). The Raman fingerprints of M. smegmatis exposed to INH change substantially in comparison to the unstressed condition. Our work provides a real-time assessment of the impact of INH on the increase of lipids in these mycobacteria, which could render the cells more tolerant to antibiotics. This microfluidic platform may be used to study any microorganism and to dynamically monitor its response to different conditions and stimuli. Baron et al. describe a microfluidic system that acoustically traps live mycobacteria and acquires label-free optical measurements over time using wavelength modulated Raman spectroscopy. Using acoustically trapped live M. smegmatis, they show that under stressed conditions produced by an antibiotic, bacteria displayed an increase in lipids which could render the cells more tolerant to antibiotics. [ABSTRACT FROM AUTHOR]

Horizontal cooperation in logistics refers to several logistics service providers cooperating to accomplish common goals, which usually involves a collaborative vehicle routing problem. In this paper, we present a new collaborative vehicle routing problem with rough location (CVRPRL), which considers both the security of sharing detailed customer information and the configuration of shared resource. We utilize the rough location of the customer to replace the detailed customer location and introduce the concept of collaborative logistics sharing degree in the CVRPRL model. Subsequently, the cost allocation mechanism is designed based on an extended Shapley value, which allocates fixed costs and risks in different ways. Further, an extended ant colony optimization (EACO) algorithm is proposed to solve the CVRPRL. The EACO algorithm combines both large neighborhood search and local search strategies. Finally, we perform a series of simulation experiments to verify the effectiveness of EACO compared with other meta-heuristic algorithms. [ABSTRACT FROM AUTHOR]

Abstract With the development of the electric vehicle (EV) technology, the electric vehicle routing problems (EVRPs) have become a current focus for research. Although customer demands in the logistics service industry are often uncertain during the route planning stage, these demands have seldom been discussed in the existing literature on the EVRPs. This study presents an electric vehicle battery swap station (BSS) location-routing problem with stochastic demands, with the aim to determine a minimum cost scheme including the optimal number and location of BSSs with an optimal route plan based on stochastic customer demands. Furthermore, the classical recourse policy and preventive restocking policy are extended by considering the influences of both battery and vehicle capacity simultaneously. Subsequently, the concept of Pareto optimality is applied to the EVRP to expedite the selection of BSS sequences. To solve such a hybrid problem, a hybrid variable neighborhood search (HVNS) algorithm is proposed, which integrates the binary particle swarm optimization and variable neighborhood search to solve the location and routing problems interactively. In experimental studies, the HVNS is compared to five heuristic algorithms to verify its performance. Highlights • Presented a novel stochastic location-routing model in EV transportation. • Extended two traditional recourse policies to deal with EV's route failure. • Proposed a hybrid VNS algorithm to solve the presented problem. • Performed experiments to verify the effectiveness of the proposed algorithm. [ABSTRACT FROM AUTHOR]

The spatial coherence of organic light-emitting diodes (OLEDs) is an important parameter that has gained little attention to date. Here, we present a method for making quantitative measurements of the spatial coherence of OLEDs using a Young's double-slit experiment. The usefulness of the method is demonstrated by making measurements on a range of OLEDs with different emitters (iridium and europium complexes) and architectures (bottom and top emitting) and the fringe visibility is further manipulated by gratings embedded in external diffractive optical elements. Based on the experiments and simulation of the results, we quantitatively determine the spatial coherence lengths of several OLEDs and find them to be a few micrometers. A 60% increase in the spatial coherence length was observed when using a narrow bandwidth emitter and a metal-coated grating. [ABSTRACT FROM AUTHOR]

Determining the identity of cells of the immune system usually involves destructive fixation and chemical staining, or labeling with fluorescently labeled antibodies recognising specific cell surface markers. Completely label-free identification would be a significant advantage in conditions where untouched cells are a priority. We demonstrate here the use of Wavelength Modulated Raman Spectroscopy, to achieve label-free identification of purified, unfixed and untouched populations of major immune cell subsets isolated from healthy human donors. Using this technique we have been able to distinguish between CD4+ T lymphocytes, CD8+ T lymphocytes and CD56+ Natural Killer cells at specificities of up to 96%. Additionally, we have been able to distinguish between CD303+ plasmacytoid and CD1c+ myeloid dendritic cell subsets, the key initiator and regulatory cells of many immune responses. This demonstrates the ability to identify unperturbed cells of the immune system, and opens novel opportunities to analyse immunological systems and to develop fully label-free diagnostic technologies. [ABSTRACT FROM AUTHOR]

We experimentally create a 'perfect' vortex beam, which has a uniform ring profile and fixed radius. In contrast with other vortex fields, the intensity profile is independent of its topological charge. We then correct this field in situ using a single trapped particle as a probe. This results in a constant angular velocity for the particle regardless of its position on the beam circumference. [ABSTRACT FROM AUTHOR]

• Investigated a large-scale heterogeneous container loading problem. • Considered weight limits and suspension constraints in this problem. • Integrated ALNS and heuristic packing algorithm for the presented problem. • Performed experiments on real-world instances. • Verified the superiority of HALNS by comparing with existing methods. This paper aims to solve the large-scale heterogeneous container loading problem (HCLP), which is an extensive form of the multiple container loading problem, in a limited time. The target is to choose a set of containers of different sizes to accommodate all products and minimize the wasted space rate. Although the heterogeneous container selection problem is a general problem in the logistics industry, few related studies have been conducted. This study also considers some practical constraints, such as weight limits and suspension constraints. A hybrid adaptive large neighborhood search (HALNS) algorithm, which includes a set of original destroy-repair operators, especially for heterogeneous container selection problems, and integrates a heuristic packing algorithm, is proposed to solve the problem in an acceptable time. To verify the efficiency of the proposed algorithm, computational experiments on real-world instances from a multinational logistics company are performed, and the results are compared with those of other existing algorithms. The results indicate that the proposed algorithm outperforms other algorithms for the HCLP. [ABSTRACT FROM AUTHOR]

The existence of a spatial correlation singularity or a ring dislocation in the spatial coherence function when a vortex is present has been demonstrated recently. Here, we investigate how the spatial correlation singularity is affected by both the radial and azimuthal mode indices (p, ℓ) in a partially coherent light field. Theoretically, we find that the spatial correlation singularity may exist even in a non-vortex beam (ℓ = 0) due to the radial index. Numerical simulations show the number of ring dislocations in the far-field cross correlation function is equal to 2p + |ℓ| for the low coherence cases. This is confirmed by our experimental results. This phenomenon may occur in any partially coherent vortex wave. [ABSTRACT FROM AUTHOR]

Stress granules (SGs), formed by untranslated messenger ribonucleoproteins (mRNPs) during cellular stress in eukaryotes, have been linked to flavivirus interference without clear understanding. This study reveals the role of Zika virus (ZIKV) NS2B as a scaffold protein mediating interaction between protein phosphatase 1α (PP1α) and eukaryotic initiation factor 2α (eIF2α). This interaction promotes eIF2α dephosphorylation by PP1α, inhibiting SG formation. The NS2B-PP1α complex exhibits remarkable stability, resisting ubiquitin-induced degradation and amplifying eIF2α dephosphorylation, thus promoting ZIKV replication. In contrast, the NS2BV35A mutant, interacting exclusively with eIF2α, fails to inhibit SG formation, resulting in reduced viral replication and diminished impact on brain organoid growth. These findings reveal PP1α's dual role in ZIKV infection, inducing interferon production as an antiviral factor and suppressing SG formation as a viral promoter. Moreover, we found that NS2B also serves as a versatile mechanism employed by flaviviruses to counter host antiviral defenses, primarily by broadly inhibiting SG formation. This research advances our comprehension of the complex interplay in flavivirus-host interactions, offering potential for innovative therapeutic strategies against flavivirus infections. Author summary: Zika virus (ZIKV) belongs to vector-borne flaviviruses, associated with Guillain-Barré syndrome and neonatal microcephaly. Stress granules (SGs) which forms under stress in eukaryotic cells can interfere with viral replication. Although previous studies have indicated that ZIKV can interfere with SG formation, the precise molecular mechanism and function remain unclear. Exploration of the mechanisms by which ZIKV antagonizes SG formation may help to identify new antiviral approaches. Here, we found that ZIKV NS2B bridge PP1α and eIF2α to promote eIF2α dephosphorylation and SG inhibition. Meanwhile, Reciprocal inhibition of K48-linked ubiquitination stabilized NS2B and PP1α to further facilitate the dephosphorylation of eIF2α and the concurrent inhibition of SG formation. Conversely, we demonstrated that a mutant form of NS2B, NS2BV35A, which interacted solely with eIF2α and not PP1α, attenuated virus replication and mitigated neurotoxicity in brain organoid development. The NS2B-PP1α-eIF2α axis reveals a novel biological function of NS2B as a general mechanism for flavivirus evasion of host antiviral effects, providing new therapeutic strategies for antiviral study. [ABSTRACT FROM AUTHOR]

• Investigated a novel electric VRP considering uncertain environment. • Proposed a fuzzy optimization model with fuzzy simulation to the presented problem. • Integrated ALNS algorithm and VND algorithm to solve the presented model. • Performed experiments to verify the effectiveness of the proposed algorithm. As fuel prices increase and emission regulations become increasingly strict, electric vehicles have been used in various logistics distribution activities. Most studies have focused on the electric vehicle routing problem under a deterministic environment, neglecting the effects of uncertain factors in practical logistics distribution. Thus, a novel fuzzy electric vehicle routing problem with time windows and recharging stations (FEVRPTW) is investigated in this study, and a fuzzy optimization model is established based on credibility theory for this problem. In the presented model, fuzzy numbers are used to denote the uncertainties of service time, battery energy consumption, and travel time. Moreover, the partial recharge is allowed under the uncertain environment. To solve the model, an adaptive large neighborhood search (ALNS) algorithm enhanced with the fuzzy simulation method is proposed. In the proposed ALNS algorithm, four new removal algorithms are designed and integrated for addressing the FEVRPTW. To further improve the algorithmic performance, the variable neighborhood descent algorithm is embedded into the proposed ALNS algorithm and five local search operators are applied. The experiments were conducted to verify the effectiveness of the proposed ALNS algorithm for solving the presented model. [ABSTRACT FROM AUTHOR]

Task scheduling is important in cloud manufacturing because of customers' increasingly individualized demands. However, when various changes occur, a previous optimal schedule may become non-optimal or even infeasible owing to the uncertainty of the real manufacturing environment where dynamic task arrival over time is a vital source. In this paper, we propose a novel collaborative task scheduling (CTS) model dealing with new task arrival which considers multi-supply chain collaboration. We present an improved multi-population biogeography-based optimization (IMPBBO) algorithm that uses a matrix-based solution representation and integrates the multi-population strategy, local search for the best solution, and the collaboration mechanism, for determining the optimal schedule. A series of experiments are conducted for verifying the effectiveness of the IMPBBO algorithm for solving the CTS model by comparing it with five other algorithms. The experimental results concerning average best values obtained by the IMPBBO algorithm are better than that obtained by comparison algorithms for 41 out of 45 cases, showing its superior performance. Wilcoxon-test has been employed to strengthen the fact that IMPBBO algorithm performs better than five comparison algorithms. [ABSTRACT FROM AUTHOR]

Population aging and a shortage of medical resources are driving the growth of home health care (HHC). Many HHC studies have focused on developing an optimal routing plan to reduce operation costs; however, few have addressed the attendant long-term sustainability concerns, such as HHC center locations and carbon emissions from transportation. This paper proposes a home health care location-routing problem with a mixed fleet of electric and conventional vehicles that considers battery swapping stations. It aims to simultaneously determine the locations of HHC centers, the scheduling of caregivers with respect to skill requirements, and a routing plan for a mixed fleet under specific time windows, load capacities, synchronized visits, and driving ranges. To address this problem, the paper proposes a novel competitive simulated annealing (CSA) algorithm in which a series of problem-specific effective local search operators expand the solution space of the CSA algorithm, with a competitive mechanism to adaptively adjust these operators to accelerate convergence speed and improve exploration ability. To enhance the exploitation ability, it employs a modified simulated annealing algorithm with a heating strategy and variable neighborhood descent. Comparison experiments are conducted to verify the superior performance of the CSA algorithm in comparison to existing algorithms and the Gurobi solver. Finally, sensitivity analyses are performed on key factors and provide some new managerial insights. [ABSTRACT FROM AUTHOR]

We previously found that, among human parainfluenza virus type 3 (HPIV3) proteins, the interaction of nucleoprotein (N) and phosphoprotein (P) provides the minimal requirement for the formation of cytoplasmic inclusion bodies (IBs), which are sites of RNA synthesis, and that acetylated α-tubulin enhances IB fusion and viral replication. In this study, using immunoprecipitation and mass spectrometry assays, we determined that vimentin (VIM) specifically interacted with the N–P complex of HPIV3, and that the head domain of VIM was responsible for this interaction, contributing to the inhibition of IB fusion and viral replication. Furthermore, we found that VIM promoted the degradation of α-tubulin acetyltransferase 1 (α-TAT1), through its head region, thereby inhibiting the acetylation of α-tubulin, IB fusion, and viral replication. In addition, we identified a 20-amino-acid peptide derived from the head region of VIM that participated in the interaction with the N–P complex and inhibited viral replication. Our findings suggest that VIM inhibits the formation of HPIV3 IBs by downregulating α-tubulin acetylation via enhancing the degradation of α-TAT1. Our work sheds light on a new mechanism by which VIM suppresses HPIV3 replication. Author summary: Human parainfluenza virus type 3 (HPIV3) is one of the most common pathogenic respiratory RNA viruses, and its replication sites, known as inclusion bodies (IBs), which are formed by nucleoprotein (N) and phosphoprotein (P), are critical in viral RNA synthesis. Exploration of the mechanisms by which viral inclusion bodies are formed in cells may help to identify new antiviral approaches. Our aim was to identify cellular proteins with regulatory effects on the formation of viral IBs. The formation of HPIV3 IBs relies on acetylated α-tubulin, and we found that VIM negatively regulated α-tubulin acetylation by enhancing the degradation of α-TAT1, thereby suppressing the fusion and maturation of viral IBs. Based on the interaction between IBs and VIM, the peptide derived from amino-acid residues 61 to 80 in the VIM head region exerted antiviral effects, providing a new direction for our antiviral study. [ABSTRACT FROM AUTHOR]

The final stage of Ebola virus (EBOV) replication is budding from host cells, where the matrix protein VP40 is essential for driving this process. Many post-translational modifications such as ubiquitination are involved in VP40 egress, but acetylation has not been studied yet. Here, we characterize NEDD4 is acetylated at a conserved Lys667 mediated by the acetyltransferase P300 which drives VP40 egress process. Importantly, P300-mediated NEDD4 acetylation promotes NEDD4-VP40 interaction which enhances NEDD4 E3 ligase activity and is essential for the activation of VP40 ubiquitination and subsequent egress. Finally, we find that Zaire ebolavirus production is dramatically reduced in P300 knockout cell lines, suggesting that P300-mediated NEDD4 acetylation may have a physiological effect on Ebola virus life cycle. Thus, our study identifies an acetylation-dependent regulatory mechanism that governs VP40 ubiquitination and provides insights into how acetylation controls EBOV VP40 egress. Author summary: Ebola virus (EBOV) is one of the deadliest pathogens, causing fatal hemorrhagic fever diseases in humans and primates. In this study, we find that P300-mediated NEDD4 acetylation facilitates EBOV egress. Acetylation promotes NEDD4-VP40 interactions which enhances NEDD4 E3 ligase activity and is essential for the activation of VP40 ubiquitination and subsequent egress. This study implies that inhibitory effect of acetylation can be regarded as an attractive candidate of drug target for the treatment of Ebola virus disease. [ABSTRACT FROM AUTHOR]

The effect of Bi2O3‐B2O3‐SiO2‐ZnO glass (denoted as BBSZ) content on the densification, microstructure, and electrical properties of ZnO‐based multilayer varistors (MLVs) has been investigated. In ZnO‐based (MLVs), BBSZ acted as a promoter in ZnO grain growth, which increased the grain size of ZnO, resulting in a decrease in breakdown voltage. Results showed that the adding of BBSZ glass had an improvement in densification and overall properties of MLVs. It was found that a maximum value of 5.45 g/cm3 of the bulk density and the optimum properties (V1mA = 27.28 V, α = 40.33, IL = 0.75 μA, Ip = 35 A) of ZnO‐based MLVs were achieved with 3 wt% BBSZ glass sintered at 925°C for 3 hours. [ABSTRACT FROM AUTHOR]

The matrix (M) proteins of paramyxoviruses bind to the nucleocapsids and cytoplasmic tails of glycoproteins, thus mediating the assembly and budding of virions. We first determined the budding characterization of the HPIV3 Fusion (F) protein to investigate the assembly mechanism of human parainfluenza virus type 3 (HPIV3). Our results show that expression of the HPIV3 F protein alone is sufficient to initiate the release of virus-like particles (VLPs), and the F protein can regulate the VLP-forming ability of the M protein. Furthermore, HPIV3F-Flag, which is a recombinant HPIV3 with a Flag tag at the C-terminus of the F protein, was constructed and recovered. We found that the M, F, and hemagglutinin-neuraminidase (HN) proteins and the viral genome can accumulate in lipid rafts in HPIV3F-Flag-infected cells, and the F protein mainly exists in the form of F1 in VLPs, lipid rafts, and purified virions. Furthermore, the function of cholesterol in the viral envelope and cell membrane was assessed via the elimination of cholesterol by methyl-β-cyclodextrin (MβCD). Our results suggest that the infectivity of HPIV3 was markedly reduced, due to defective internalization ability in the absence of cholesterol. These results reveal that HPIV3 might assemble in the lipid rafts to acquire cholesterol for the envelope of HPIV3, which suggests the that disruption of the cholesterol composition of HPIV3 virions might be a useful method for the design of anti-HPIV3 therapy. [ABSTRACT FROM AUTHOR]

Abstract Secretory IgA (SIgA) antibody is unique for its capability to transit through epithelial cells by transcytosis and thus has opportunities and probabilities to interact with all viral components during viral replication which may result in the inhibition of viral replication intracellularly. Here, we report a novel IgA mAb 1D11-IgA against phosphoprotein (P) of measles virus (MV), which is able to interact specifically with P in MV infected Vero-pIgR cells grown in a two-chamber transwell system. The binding epitope of 1D11-IgA involves a key residue proline 23 in P protein, which is among the α-molecular recognition element (α-MoRE) of P and critical for N0-P complex. The antibody appears to block P to interact with N in P-N complex and thus may inhibit the function of viral RdRp complex, which results in decreased synthesis of viral genome RNA and mRNA. Our data together demonstrate that IgA is able to interact with viral phosphoprotein intraepithelial cells and neutralize viral replication by interrupting formation of P-N complex and function of RdRp. The findings highlight that IgA has a unique anti-viral activity by targeting viral conserved components critical for viral replication, which serves as a proof-of-concept assessment of the druggability of mononegavirales P-N interfaces. Highlights • A novel IgA mAb 1D11-IgA targeting on the α-MoRE of viral phosphoprotein inhibits measles virus replication intracellularly. • The 1D11-IgA inhibits measles virus replication by interrupting P-N complex formation during its transcytosis. • The study highlights the α-MoRE of P is a very specific and promising antiviral target for antiviral drug screen. [ABSTRACT FROM AUTHOR]

The phosphoprotein (P) of human parainfluenza virus type 3 (HPIV3) plays a pivotal role in viral RNA synthesis, which interacts with the nucleoprotein (N) to form a soluble N 0 -P complex (N 0 , free of RNAs) to prevent the nonspecific RNA binding and illegitimate aggregation of N. Functional regions within P have been studied intensively. However, the precise site (s) within P directly involved in N 0 -P interaction still remains unclear. In this study, using a series of deleted and truncated mutants of P of HPIV3, we demonstrate that amino-terminal 40 amino acids (aa) of P restrict and regulate N 0 -P interaction. Furthermore, using in vivo HPIV3 minigenome replicon assay, we identify a critical P mutant (P A28P ) located in amino-terminal 40 aa, which fails to support RNA synthesis of HPIV3 minigenome replicon. Although P A28P maintains an enhanced N-P interaction, it is unable to form N 0 -P complex and keep N soluble, thus, resulting in aggregation and functional abolishment of N-P complex. Moreover, we found that recombinant HPIV3 with mutation of A28P in P failed to be rescued. Taken together, we identified a residue within the extreme amino-terminus of P, which plays a critical role in restricting the excessively N-P interaction and keeping a functional N 0 -P complex formation. [ABSTRACT FROM AUTHOR]

Viral invasion triggers the activation of the host antiviral response. Besides the innate immune response, stress granules (SGs) also act as an additional defense response to combat viral replication. However, many viruses have evolved various strategies to suppress SG formation to facilitate their own replication. Here, we show that viral mRNAs derived from human parainfluenza virus type 3 (HPIV3) infection induce SG formation in an eIF2α phosphorylation- and PKR-dependent manner in which viral mRNAs are sequestered and viral replication is inhibited independent of the interferon signaling pathway. Furthermore, we found that inclusion body (IB) formation by the interaction of the nucleoprotein (N) and phosphoprotein (P) of HPIV3 correlated with SG suppression. In addition, co-expression of P with NL478A (a point mutant of N, which is unable to form IBs with P) or with NΔN10 (lacking N-terminal 10 amino acids of N, which could form IBs with P but was unable to synthesize or shield viral RNAs) failed to inhibit SG formation, suggesting that inhibition of SG formation also correlates with the capacity of IBs to synthesize and shield viral RNAs. Therefore, we provide a model whereby viral IBs escape the antiviral effect of SGs by concealing their own newly synthesized viral RNAs and offer new insights into the emerging role of IBs in viral replication. [ABSTRACT FROM AUTHOR]

Human parainfluenza virus type 3 is one of the main causes of lower respiratory illness in newborns and infants. The role of the matrix protein (M) in viral budding is extensively studied, but the effect of M on viral replication remains to be determined. Using an HPIV3 minigenome assay, we found that M reduced HPIV3 mingenome-encoded reporter activity even though it had an unspecific effect on the expression of cellular genes. Furthermore, the inhibition effect of M on viral RNA synthesis was proven to be independent of its virus-like particles (VLPs)' release ability. A VLP's defective mutant (ML302A) decreased the expression of minigenome reporter as wild type M did. Using an immunofluorescence assay, we found that M weakened the formation of inclusion bodies (IBs), although it did not co-localize with the IBs. Moreover, using another mutant, ML305A , which is defective in M-nucleoprotein (N) interaction, we found that ML305A had no effect on reporter activity and IB formation as the wild type of M did. Taken together, we conclude that M reduces the replication of HPIV3 and IB formation by M-N interaction. [ABSTRACT FROM AUTHOR]

Stress granules (SGs) contain stalled messenger ribonucleoprotein complexes and are related to the regulation of mRNA translation. Picornavirus infection can interfere with the formation of SGs. However, the detailed molecular mechanisms and functions of picornavirus-mediated regulation of SG formation are not clear. Here, we found that the 2A protease of a picornavirus, EV71, induced atypical stress granule (aSG), but not typical stress granule (tSG), formation via cleavage of eIF4GI. Furthermore, 2A was required and sufficient to inhibit tSGs induced by EV71 infection, sodium arsenite, or heat shock. Infection of 2A protease activity-inactivated recombinant EV71 (EV71-2AC110S) failed to induce aSG formation and only induced tSG formation, which is PKR and eIF2α phosphorylation-dependent. By using a Renilla luciferase mRNA reporter system and RNA fluorescence in situ hybridization assay, we found that EV71-induced aSGs were beneficial to viral translation through sequestering only cellular mRNAs, but not viral mRNAs. In addition, we found that the 2A protease of other picornaviruses such as poliovirus and coxsackievirus also induced aSG formation and blocked tSG formation. Taken together, our results demonstrate that, on one hand, EV71 infection induces tSG formation via the PKR-eIF2α pathway, and on the other hand, 2A, but not 3C, blocks tSG formation. Instead, 2A induces aSG formation by cleaving eIF4GI to sequester cellular mRNA but release viral mRNA, thereby facilitating viral translation. [ABSTRACT FROM AUTHOR]

The ability to identify and characterise individual cells of the immune system under label-free conditions would be a significant advantage in biomedical and clinical studies where untouched and unmodified cells are required. We present a multi-modal system capable of simultaneously acquiring both single point Raman spectra and digital holographic images of single cells. We use this combined approach to identify and discriminate between immune cell populations CD4+ T cells, B cells and monocytes. We investigate several approaches to interpret the phase images including signal intensity histograms and texture analysis. Both modalities are independently able to discriminate between cell subsets and dual-modality may therefore be used a means for validation. We demonstrate here sensitivities achieved in the range of 86.8% to 100%, and specificities in the range of 85.4% to 100%. Additionally each modality provides information not available from the other providing both a molecular and a morphological signature of each cell. [ABSTRACT FROM AUTHOR]

China is the largest country in grain production and consumption with the greatest emission of greenhouse gas (GHG) in the world. To learn the historical changes and regional differences in GHG emission of the major grain crop production can provide important references to the mitigation of GHG emission in the country and other similar countries. Therefore, we quantified the carbon footprints of rice ( Oryza sativa L.), wheat ( Triticumaestivum L.) and maize ( Zea mays L.) production based on agricultural inputs in the country over the period of 1978–2012. The results showed that area-scaled carbon footprint of rice, wheat and maize production increased gradually from 1286, 937 and 895 kg carbon dioxide equivalent (CO 2 -eq) ha −1 in 1978–2682, 2978 and 2294 kg CO 2 -eq ha −1 in 2012, respectively, and the average increase rates for the three crops were correspondingly 41, 60 and 41 kg CO 2 -eq ha −1 a −1 . During the last ten years, however, all the average yield-scaled carbon footprints of rice, wheat and maize decreased by 0.11 kg CO 2 -eq kg −1 10a −1 . Chemical fertilizer contributed the largest ratio to the total GHG emission by 66, 68 and 76% for rice, wheat and maize, respectively. Significant increasing contribution ratio to the carbon footprint was found in the electricity used for crop irrigation. There were large differences in both the area- and yield-scaled carbon footprints among the cropping regions with the lowest GHG emission occurring in the major cropping regions. Our findings indicate a great potential to reduce GHG emission of grain production through optimizing the spatial layout of cropping region and enhancing chemical fertilizer use efficiency. Further efforts also need to be made on water-saving irrigation to decrease the electricity usage. [ABSTRACT FROM AUTHOR]

Enterovirus 71 (EV71) belonging to the Picornaviridae family is considered the most frequently detected causative agent in hand-foot-and-mouth disease (HFMD) and is a serious threat to public health in the Asia-Pacific region. There are currently no approved vaccines or effective drugs for EV71. In this study, using recombinant vesicular stomatitis virus (rVSV) expressing viral VP1 protein (mVP1) of EV71 as a control, we generated two types of rVSVs that can form EV71 virus-like particles (VLPs). First, we co-infected two rVSVs singly expressing P1 (mP1) and 3CD (m3CD) of EV71. Second, we inserted P1 and 3CD into one VSV backbone to generate an rVSV expressing P1 and 3CD together (mP1-3CD). When P1 and 3CD were expressed in the cells either co-infected with mP1 and m3CD (mP1/m3CD) or infected with mP1-3CD, P1 was cleaved by 3CD and produced VP1, VP3, and VP0 to form VLPs. Furthermore, mice immunized with mP1/m3CD or mP1-3CD showed higher humoral and cellular immunity responses than mice immunized with mVP1. Finally, the rVSVs expressing the EV71 proteins were evaluated in mice to determine their potential to protect against a lethal EV71 virus challenge, and among all the rVSVs, the mP1-3CD was shown to be the most promising vaccine candidate for EV71 protection. [ABSTRACT FROM AUTHOR]

The functional template for the transcription and replication of vesicular stomatitis virus (VSV) is genomic RNA encapsidated by nucleocapsid (N) protein. Previous studies showed that the amino acid R7 in the N-terminal arm of N is involved in N–N interaction in the N-RNA complex. In our study, the recombinant virus with mutation of R7A (rVSV R7A ) in N was recovered, and the replication level of passage 1 (P1) of rVSV R7A was 1000 times lower than that of wild-type rVSV at 37 °C. After eight passages, the replication level of P8 of rVSV R7A with two second-site mutations in the genome (T242 P in N protein and U7-U8 in G-L gene junction) was significantly higher than that of P1. Furthermore, we demonstrate that the mutation of either T242 P or U7-U8 can compensate the effect caused by the mutation of R7A on the replication of rVSV R7A . Therefore, we conclude that two second-site mutations both can compensated the engineered mutation of R7A in VSV N protein. [ABSTRACT FROM AUTHOR]

The nucleoprotein (N) of vesicular stomatitis virus (VSV) plays a central role in transcription and replication by encapsidating genome RNA to form a nucleocapsid as the template for the RNA synthesis. Using minigenome system we evaluated the roles of 21 amino acids of the N-terminal arm of N in forming functional N–RNA templates and found that three triple-amino-acid substitutions (TVK4-6A3, RII7-9A3, and VIV13-15A3) and one single-amino-acid substitution (R7A) resulted in RNA synthesis loss. But all the mutants maintain the ability to oligomerize N, interact with P, and encapsidate viral RNA for template formation. Further analysis showed that the nucleocapsid formed by these mutants failed to protect RNA from nuclease digestion. Then, we found that only recombinant viruses containing R7A could be recovered. Our results show that the several amino acids within the N-terminal arm of N contribute to the template function beyond its role in RNA encapsidation and viral growth. [ABSTRACT FROM AUTHOR]

The phosphoprotein (P) of vesicular stomatitis virus (VSV) plays essential roles in viral RNA synthesis. It associates with nascent nucleoprotein (N) to form N0-P (free of RNAs), thereby preventing the N from binding to cellular RNAs and maintaining the N in a viral genomic RNA encapsidation-competent form for transcription and replication. The contributions of phosphorylation of P to transcription and replication have been studied intensively, but a concrete mechanism of action still remains unclear. In this study, using a VSV minigenome system, we demonstrated that a mutant of P lacking N-terminal phosphorylation (P3A), in which the Nterminal phosphate acceptor sites are replaced with alanines (S60/A, T62/A, and S64/A), does not support transcription and replication. However, results from protein interaction assays showed that P3A self-associates and interacts with N and the large protein (L) as efficiently as P does. Furthermore, purified recombinant P3A from Sf21 cells supported transcription in an in vitro transcription reconstitution assay. We also proved that P3A is not distributed intranuclearly in vivo. CsCl gradient centrifugation showed that P3A is incapable of preventing N from binding to cellular RNAs and therefore prevents functional template formation. Taken together, our results demonstrate that N-terminal phosphorylation is indispensable for P to prevent N from binding to nonviral RNAs and to maintain the N-specific encapsidation of viral genomic RNA for functional template formation. [ABSTRACT FROM AUTHOR]

Abstract: Due to the toxicity of dioxins, furans and heavy metals, there is a growing environmental concern on municipal solid waste incinerator (MSWI) fly ash in China. The purpose of this study is directed towards the volume-reduction of fly ash without any additive by thermal plasma and recycling of vitrified slag. This process uses extremely high-temperature in an oxygen-starved environment to completely decompose complex waste into very simple molecules. For developing the proper plasma processes to treat MSWI fly ash, a new crucible-type plasma furnace was built. The melting process metamorphosed fly ash to granulated slag that was less than 1/3 of the volume of the fly ash, and about 64% of the weight of the fly ash. The safety of the vitrified slag was tested. The properties of the slag were affected by the differences in the cooling methods. Water-cooled and composite-cooled slag showed more excellent resistance against the leaching of heavy metals and can be utilized as building material without toxicity problems. [Copyright &y& Elsevier]

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is currently a global pandemic. CoVs are known to generate negative subgenomes (subgenomic RNAs [sgRNAs]) through transcription-regulating sequence (TRS)-dependent template switching, but the global dynamic landscapes of coronaviral subgenomes and regulatory rules remain unclear. Here, using next-generation sequencing (NGS) short-read and Nanopore long-read poly(A) RNA sequencing in two cell types at multiple time points after infection with SARS-CoV-2, we identified hundreds of template switches and constructed the dynamic landscapes of SARS-CoV-2 subgenomes. Interestingly, template switching could occur in a bidirectional manner, with diverse SARS-CoV-2 subgenomes generated from successive template-switching events. The majority of template switches result from RNA-RNA interactions, including seed and compensatory modes, with terminal pairing status as a key determinant. Two TRS-independent template switch modes are also responsible for subgenome biogenesis. Our findings reveal the subgenome landscape of SARS-CoV-2 and its regulatory features, providing a molecular basis for understanding subgenome biogenesis and developing novel anti-viral strategies. [Display omitted] • Dynamic subgenome landscapes of SARS-CoV-2 in two host cells are constructed • Bidirectional and successive template switching diversify sgRNA biogenesis • Several key determinants governing template switching efficacy are discovered • Canonical TRS-independent RNA-RNA interaction mediates template switches Wang et al. construct dynamic landscapes of SARS-CoV-2 subgenomic RNAs (sgRNAs) by using integrated poly(A) RNA sequencing at various time points after infection. Computational analyses reveal novel modes of viral sgRNA biogenesis and decode regulatory features, including several key determinants governing the efficacy of template switching in coronaviral RNA transcription. [ABSTRACT FROM AUTHOR]

Fiber optic sensors are widely used in environmental, biological and chemical sensing. Due to the demanding environmental conditions in which they can be used, there is a risk of damaging the sensor measurement head placed in the measuring field. Sensors using nanolayers deposited upon the fiber structure are particularly vulnerable to damage. A thin film placed on the surface of the fiber end-face can be prone to mechanical damage or deteriorate due to unwanted chemical reactions with the surrounding agent. In this paper, we investigated a sensor structure formed with a Zinc Oxide (ZnO) coating, deposited by Atomic Layer Deposition (ALD) on the tip of a single-mode fiber. A nanocrystalline diamond sheet (NDS) attached over the ZnO is described. The diamond structure was synthesized in a Microwave Plasma Assisted Chemical Vapor Deposition System. The deposition processes of the nanomaterials, the procedure of attaching NDS to the fiber end-face covered with ZnO, and the results of optical measurements are presented. [ABSTRACT FROM AUTHOR]