Fate characteristics and risk quantification of cyflumetofen from tomato cultivation to processing based on large-scale applications.

Elucidating the fate characteristics of cyflumetofen and its main metabolite 2-TFMBA in tomato from cultivation to processing is crucial for safeguarding the environment and humans from hazardous effects. Cyflumetofen and 2-TFMBA could exist stably in tomato matrices for at least 343 days under frozen and dark conditions according to UHPLC-MS/MS, with a limit of quantitation of 0.001 mg/kg and retention time within 2.12 min. The occurrence, dissipation, and concentration variation of cyflumetofen were reflected by original depositions of 0.02-0.44 mg/kg, half-lives of 1.7-7.2 days, and terminal magnitudes of 0.005-0.30 mg/kg, respectively, with various influencing factors, e.g., climate conditions and tomato cultivars. Additionally, 13.5-59.3% of cyflumetofen was metabolized to 2-TFMBA, showing significant toxicological effects ranging from cultivation to processing. When the concentration decreased by 0.06 mg/kg, cyflumetofen was effectively removed by peeling, while washing was the recommended method for removing 2-TFMBA with a processing factor of 0.70. The comparative dietary risks of sum cyflumetofen were assessed for all life cycle populations using deterministic and probabilistic models. The risk quotients decreased to 1.3-4.8 times during the preparation of home canning tomato paste. Despite the low exposure risk, the potential health hazards of sum cyflumetofen should be considered, given its ubiquity and cumulative effects.

Unique and generic crossed metabolism in response to four sub-lethal environmental stresses in the oriental fruit fly, Bactrocera dorsalis Hendel.

Bactrocera dorsalis is a well-known invasive pest that causes considerable ecological and economic losses worldwild. Although it has a wide environmental tolerance, few studies have reported its mechanism of adaptation to multiple sub-lethal environmental stresses. In this study, 38, 41, 39 and 34 metabolites changed significantly in B. dorsalis under four sub-lethal stresses (heat, cold, desiccation and hypoxia), as found by the metabolomic method. Therein, lactic acid and pyruvic acid were induced, whereas metabolites in the tricarboxylic acid (TCA) cycle such as citric acid, α-ketoglutarate acid, malic acid and fumaric acid were reduced under at least one of the stresses. Enzyme activity and quantitative polymerase chain reaction (qPCR) analyses verified the repression of pyruvic acid proceeding into the TCA cycle. In addition, the levels of several cryoprotectants and membrane fatty acids in B. dorsalis were altered. The findings indicated that B. dorsalis has evolved shared metabolic pathways to adapt to heat, hypoxia and desiccation stresses, such as reducing energy consumption by activating the anaerobic glycolytic metabolism. Cryoprotectants and membrane fatty acids were produced to improve the efficiency of stress resistance. This study revealed the unique and generic crossed physiological mechanism of insects to adapt to various environmental stresses.

Risk assessment framework for pine wilt disease: Estimating the introduction pathways and multispecies interactions among the pine wood nematode, its insect vectors, and hosts in China.

Pine wilt disease (PWD), caused by the pine wood nematode (PWN, Bursaphelenchus xylophilus), a destructive, invasive forest pathogen, poses a serious threat to global pine forest ecosystems. The global invasion of PWN has been described based on three successive phases, introduction, establishment, and dispersal. Risk assessments of the three successive PWN invasion phases can assist in targeted management efforts. Here, we present a risk assessment framework to evaluate the introduction, establishment, and dispersal risks of PWD in China using network analysis, species distribution models, and niche concepts. We found that >88 % of PWN inspection records were from the United States, South Korea, Japan, Germany, and Mexico, and 94 % of interception records were primarily from the Jiangsu, Shanghai, Shandong, Tianjin, and Zhejiang ports. Based on the nearly current climate, the areas of PWN overlap with its host Pinus species were primarily distributed in southern, eastern, Yangtze River Basin, central, and northeastern China regions. Areas of PWN overlap with its insect vector Monochamus alternatus were primarily distributed in southern, eastern, Yangtze River Basin, central, and northeastern China regions, and those of PWN overlap with the insect vector Monochamus saltuarius were primarily distributed in eastern and northeastern China. The niche between PWN and the insect vector M. alternatus was the most similar (0.68), followed by that between PWN and the insect vector M. saltuarius (0.47). Climate change will increase the suitable probabilities of PWN and its two insect vectors occurring at high latitudes, further increasing their threat to hosts in northeastern China. This risk assessment framework for PWD could be influential in preventing the entry of the PWN and mitigating their establishment and dispersal risks in China. Our study provides substantial clues for developing a framework to improve the risk assessment and surveillance of biological invasions worldwide.

Palmitoleic acid as a coordinating molecule between the invasive pinewood nematode and its newly associated fungi.

Symbiotic microorganisms are ubiquitous on the body surface or internal tissues of invertebrates, providing them with benefits. Developing symbiotic relationships requires synchronization of developmental stages and physical proximity of partners. Therefore, the identification of metabolites that coordinate the reproduction of symbiotic partners is essential. This study demonstrates that palmitoleic acid (C16: 1) coordinates bilateral propagation by regulating the synchronization of reproduction between the invasive pinewood nematode (PWN) and its newly associated blue-stain fungus, Sporothrix sp.1. When the PWN fed on Sporothrix sp.1, there was a significant increase in lipid metabolism gene expression and metabolite abundance. Through further investigations, it highlighted a significant enhancement in the reproduction of the PWN through direct acquisition of C16: 1, which was abundantly present in Sporothrix sp.1. Furthermore, the PWN biosynthesized C16: 1 through the involvement of the stearoyl-CoA 9-desaturase gene fat-5 and its hormone nuclear receptor nhr-80, which was clarified to promote the egg-laying capacity of females. Moreover, it is worth noting that the production of C16: 1 was significantly higher by the associated fungus Sporothrix sp.1 to enhance sporulation during the spore formation phase compared to the hypha growth phase. Thus, by coordinating the fecundity and spore production, the key lipid metabolite C16: 1 facilitates the rapid and successful colonization of a mutually beneficial symbiotic relationship between the invasive PWN and the native Sporothrix sp.1 within the host. This finding emphasizes the significant role of metabolite sharing and its function in promoting partner synchronization within symbiotic relationships.

Temperature-regulated metabolites of Serratiamarcescens inhibited reproduction of pinewood nematode Bursaphelenchus xylophilus.

The pinewood nematode Bursaphelenchus xylophilus is an invasive and destructive pathogen in forestry. Serratia marcescens AHPC29 was previously found to have nematicidal activity on B. xylophilus. The effect of AHPC29 growth temperature on B. xylophilus inhibition is unknown. Here we show that AHPC29 cultured at 15°C or 25°C, but not 37°C, inhibited B. xylophilus reproduction. Metabolomic analysis found 31 up-regulated metabolites as potential effective substances in this temperature-related difference, with five of them were tested to be effective in inhibiting B. xylophilus reproduction. Among the five metabolites, salsolinol was further verified in bacterial cultures with effective inhibition concentrations. This study found the inhibition of S. marcescens AHPC29 on B. xylophilus reproduction was temperature regulated and the differently expressed metabolites salsolinol played roles in this temperature-regulated effect, which implies the capability of S. marcescens and its metabolites as promising new agents for the management of B. xylophilus.

Health risks of dietary ethirimol exposure demand attention in China: A study of nationwide applications.

The health risks for dietary exposure after large-scale ethirimol application demand attention to protect the human population from hazardous effects. The occurrence, dissipation and terminal magnitude of ethirimol were revealed by original deposition of <0.005-28.1 mg kg-1, half-lives of 0.6-14.5 d, and terminal concentrations of  spinach > leaf lettuce > pepper > cucumber > papaya > pumpkin > stem lettuce > muskmelon > wax gourd > mango. The high-potential health risks of ethirimol should be continuously emphasized for susceptible children aged 2-13 years, especially those residing in rural areas and presenting increasing vegetable intake.

Fate and occurrence of indoxacarb during radish cultivation for multi-risk assessment.

Agrochemical indoxacarb is an important tool for selective pest control in radish that be consumed globally. A rapid and sensitive analytical method UHPLC-MS/MS was developed for tracing indoxacarb in radish leaves and roots with LOQ of 0.001 mg/kg and RT within 2 min, which were confirmed the satisfied storage stability of indoxacarb in radish matrixes with degradation rates less than 30 %. The occurrence, pharmacokinetics dissipation and concentration variation of indoxacarb were reflected by the original deposition of 2.23-4.12 mg/kg, half-lives of 2.6-8.0 d and terminal magnitude of 0.17 × 10-2-25.46 mg/kg in radish, and the influencing factors were further illustrated in terms of climate factors, crop cultivars and soil properties. The highest residues of indoxacarb were 25.46 mg/kg in leaves and 0.12 mg/kg in roots, which were higher than international maximum residue limits. A probabilistic model, as well as deterministic model, were introduced to evaluated the health risks of indoxacarb offering a better description for uncertainty. The total chronic dietary risk values of indoxacarb were 146.961-482.065 % in 12 registered crops, of which ADI % in radish was accounted for 19.8 % with risk dilution effects. The unacceptable acute dietary risks of 121.358-220.331 % were observed at 99.9th percentile, whereas the high-potential non-carcinogenic effects were observed over 90th percentile (105.035-1121.943 %). The health risks should be continuously emphasized given the increasing applications and persistent characteristics of indoxacarb, which is vital to protect the human population from hazardous effects, particularly for vulnerable children.

Neural circuit mechanisms linking courtship and reward in Drosophila males.

Courtship has evolved to achieve reproductive success in animal species. However, whether courtship itself has a positive value remains unclear. In the present work, we report that courtship is innately rewarding and can induce the expression of appetitive short-term memory (STM) and long-term memory (LTM) in Drosophila melanogaster males. Activation of male-specific P1 neurons is sufficient to mimic courtship-induced preference and memory performance. Surprisingly, P1 neurons functionally connect to a large proportion of dopaminergic neurons (DANs) in the protocerebral anterior medial (PAM) cluster. The acquisition of STM and LTM depends on two distinct subsets of PAM DANs that convey the courtship-reward signal to the restricted regions of the mushroom body (MB) γ and α/ß lobes through two dopamine receptors, D1-like Dop1R1 and D2-like Dop2R. Furthermore, the retrieval of STM stored in the MB α'/ß' lobes and LTM stored in the MB α/ß lobe relies on two distinct MB output neurons. Finally, LTM consolidation requires two subsets of PAM DANs projecting to the MB α/ß lobe and corresponding MB output neurons. Taken together, our findings demonstrate that courtship is a potent rewarding stimulus and reveal the underlying neural circuit mechanisms linking courtship and reward in Drosophila males.

A chemical signal that promotes insect survival via thermogenesis.

Cold-activated thermogenesis of brown adipose tissues (BAT) is vital for the survival of animals under cold stress and also inhibits the development of tumours. The development of small-molecule tools that target thermogenesis pathways could lead to novel therapies against cold, obesity, and even cancer. Here, we identify a chemical signal that is produced in beetles in the winter to activate fat thermogenesis. This hormone elevates the basal body temperature by increasing cellular mitochondrial density and uncoupling in order to promote beetle survival. We demonstrate that this hormone activates UCP4- mediated uncoupled respiration through adipokinetic hormone receptor (AKHR). This signal serves as a novel fat-burning activator that utilizes a conserved mechanism to promote thermogenesis not only in beetles, nematode and flies, but also in mice, protecting the mice against cold and tumor growth. This hormone represents a new strategy to manipulate fat thermogenesis.

Hypoxia-induced tracheal elasticity in vector beetle facilitates the loading of pinewood nematode.

Many pathogens rely on their insect vectors for transmission. Such pathogens are under selection to improve vector competence for their transmission by employing various tissue or cellular responses of vectors. However, whether pathogens can actively cause hypoxia in vectors and exploit hypoxia responses to promote their vector competence is still unknown. Fast dispersal of pinewood nematode (PWN), the causal agent for the destructive pine wilt disease and subsequent infection of pine trees, is characterized by the high vector competence of pine sawyer beetles (Monochamus spp.), and a single beetle can harbor over 200,000 PWNs in its tracheal system. Here, we demonstrate that PWN loading activates hypoxia in tracheal system of the vector beetles. Both PWN loading and hypoxia enhanced tracheal elasticity and thickened the apical extracellular matrix (aECM) of the tracheal tubes while a notable upregulated expression of a resilin-like mucin protein Muc91C was observed at the aECM layer of PWN-loaded and hypoxic tracheal tubes. RNAi knockdown of Muc91C reduced tracheal elasticity and aECM thickness under hypoxia conditions and thus decreasing PWN loading. Our study suggests a crucial role of hypoxia-induced developmental responses in shaping vector tolerance to the pathogen and provides clues for potential molecular targets to control pathogen dissemination.

Various parasites, bacteria and other disease-causing pathogens are transmitted by insects. A tiny worm called the pine wood nematode, for example, is spread by pine sawyer beetles which can carry up to 280,000 worms in their trachea, the network of tubes they use to breathe. This has resulted in millions of hectares of pine forests in Asia and Europe becoming infected with the deadly disease caused by the nematodes. Pine wood nematodes, as well as other pathogens, can exploit the biological processes of the insects carrying them to make the insects transmit them more effectively. Precisely how nematodes and other disease-causing agents do this is unclear. One possibility is that they reduce the amount of oxygen being supplied to the trachea ­ a phenomenon known as hypoxia ­ which occurs naturally at specific stages in the life of an insect, and during infections. To test this theory, Tang, Zhou, Koski et al. used genetics and imaging approaches to study how pine wood nematodes affect the trachea of pine sawyer beetles. The experiments found that when the nematodes infected the beetles, their trachea did indeed develop hypoxia. This, in turn, made the beetles' airways more elastic and made the layer of structure lining the trachea, known as the apical extracellular matrix, thicker. These changes increased the amount of pinewood nematodes the trachea could hold, allowing the beetle to spread more worms from tree to tree. Further experiments revealed that hypoxia in the trachea increased the levels of a protein called Muc91C in the apical extracellular matrix. When the levels of Muc91C were artificially decreased in the beetles, this made their airways less elastic and the apical extracellular matrix thinner. This work suggests that pine wood nematodes exploit the beetles' normal responses to loss of oxygen supply to make the beetles more effective at transmitting the nematodes between pine trees. Other pathogens carried by insects may also use this strategy to help increase their transmission. Further studies on the Muc91C protein may provide clues for potential drug targets to control pine wood nematodes and protect pine trees from disease.

Large-scale fate profiling of butralin between cultivated and processed garlics for multi-risk estimations.

Elaborating on the fate profiling and risk magnitude of butralin during large-scale applications was conducive to agroecosystems sustainability and dietary rationality. Occurrence, dissipation and concentration variation of butralin were elucidated from garlic cultivation to household processing by tracing UHPLC-MS/MS within 2 min, with regard to original depositions, half-lives, and terminal magnitude in typical origins of garlic. The processing factors (Pfs) of butralin were further clarified among washing, stir-frying and pickling of garlic crops, and pickling was the most effective way for butralin removal with a Pf of 0.092. A probabilistic model with Pfs was further introduced for the comprehensive risk estimations, by reduction factors of 3.1-10.9 from raw garlic crops to processed products. The short-term risks of butralin from green garlic were greater than those between garlic shoot and garlic, with the %ARfDs of 0.030 %-6.323 % from 50th to 99.9th percentiles. The long-term risks were inversely correlated to the age of the population, whose location in rural (%ADIs, 0.256 %-0.768 %) suffered more serious exposures than in urban (%ADIs, 0.231 %-0.699 %). High potential risk amplification should be continuously emphasized given the increasing applications and persistent fate of butralin, especially for vulnerable rural children.

Invasion History of the Pinewood Nematode Bursaphelenchus xylophilus Influences the Abundance of Serratia sp. in Pupal Chambers and Tracheae of Insect-Vector Monochamus alternatus.

Pine wilt disease (PWD) has caused extensive mortality in pine forests worldwide. This disease is a result of a multi-species interaction among an invasive pinewood nematode (PWN) Bursaphelenchus xylophilus, its vector Monochamus sp. beetle, and the host pine tree (Pinus sp.). In other systems, microbes have been shown to attenuate negative impacts on invasive species after the invasion has reached a certain time point. Despite that the role of PWD associated microbes involved in the PWD system has been widely studied, it is not known whether similar antagonistic "hidden microbial players" exist in this system due to the lack of knowledge about the potential temporal changes in the composition of associated microbiota. In this study, we investigated the bacteria-to-fungi ratio and isolated culturable bacterial isolates from pupal chambers and vector beetle tracheae across five sampling sites in China differing in the duration of PWN invasion. We also tested the pathogenicity of two candidate bacteria strains against the PWN-vector beetle complex. A total of 118 bacterial species belonging to 4 phyla, 30 families, and 54 genera were classified based on 16S sequencing. The relative abundance of the genus Serratia was lower in pupal chambers and tracheae in newly PWN invaded sites (<10 years) compared to the sites that had been invaded for more than 20 years. Serratia marcescens strain AHPC29 was widely distributed across all sites and showed nematicidal activity against PWN. The insecticidal activity of this strain was dependent on the life stage of the vector beetle Monochamus alternatus: no insecticidal activity was observed against final-instar larvae, whereas S. marcescens was highly virulent against pupae. Our findings improved the understanding of the temporal variation in the microbial community associated with the PWN-vector beetle complex and the progress of PWD and can therefore facilitate the development of biological control agents against PWN and its vector beetle.

Microhabitat Governs the Microbiota of the Pinewood Nematode and Its Vector Beetle: Implication for the Prevalence of Pine Wilt Disease.

Our understanding of environmental acquisition of microbes and migration-related alteration of microbiota across habitats has rapidly increased. However, in complex life cycles, such as for many parasites, exactly how these microbes are transmitted across multiple environments, such as hosts and habitats, is unknown. Pinewood nematode, the causal agent of the globally devastating pine wilt disease, provides an ideal model to study the role of microbiota in multispecies interactions because its successful host invasion depends on the interactions among its vector insects, pine hosts, and associated microbes. Here, we studied the role of bacterial and fungal communities involved in the nematode's life cycle across different micro- (pupal chamber, vector beetle, and dispersal nematodes) and macrohabitats (geographical locations). We identified the potential sources, selection processes, and keystone taxa involved in the host pine-nematode-vector beetle microbiota interactions. Nearly 50% of the microbiota in vector beetle tracheae and ~60% that of third-stage dispersal juveniles were derived from the host pine (pupal chambers), whereas 90% of bacteria of fourth-stage dispersal juveniles originated from vector beetle tracheae. Our results also suggest that vector beetles' tracheae selectively acquire some key taxa from the microbial community of the pupal chambers. These taxa will be then enriched in the dispersal nematodes traveling in the tracheae and hence likely transported to new host trees. Taken together, our findings contribute to the critical information toward a better understanding of the role of microbiota in pine wilt disease, therefore aiding the knowledge for the development of future biological control agents. IMPORTANCE Our understanding of animal microbiota acquisition and dispersal-mediated variation has rapidly increased. In this study, using the model of host pine-pinewood nematode-vector beetle (Monochamus sp.) complex, we disentangled the routes of microbial community assembly and transmission mechanisms among these different participants responsible for highly destructive pine wilt disease. We provide evidence that the microhabitat is the driving force shaping the microbial community of these participants. The microbiota of third-stage dispersal juveniles (LIII) of the nematodes collected around pupal chambers and of vector beetles were mainly derived from the host pine (pupal chambers), whereas the vector-entering fourth-stage dispersal juveniles (LIV) of the nematodes had the simplest microbiota community, not influencing vector's microbiota. These findings enhanced our understanding of the variation in the microbiota of plants and animals and shed light on microbiota acquisition in complex life cycles.

Parallel Evolution of C-Type Lectin Domain Gene Family Sizes in Insect-Vectored Nematodes.

The dispersal stage of pathogens is crucial for the successful spread and infection of their hosts. Some plant-parasitic nematodes (PPNs) have evolved specialized dispersal stages to reach healthy hosts by being carried out by insect vectors. Because gene gain and loss is a major factor contributing to the evolution of novel characteristics, it is essential to clarify the gene family characteristics among nematodes with different dispersal modes to disentangle the evolution of insect-mediated dispersal. Here, the size of the C-type lectin (CTL) family genes of insect-vectored nematodes was found to be drastically reduced compared with those of self-dispersing nematodes, whereas the diversity of their functional domains was significantly higher. The gene family sizes of vector-dispersed nematodes were only a twentieth of the size of that of a self-dispersing (i.e., without a biotic vector) nematode model Caenorhabditis elegans, and these genes were inactive during the dispersal stage. Phylogenetic analysis showed that some CTL genes of vector-borne PPNs shared higher homology to the animal parasitic nematodes compared with other PPNs. Moreover, homology modeling predicted that the CTLs of insect-vectored nematodes bear remarkable structural similarity to the lectin genes of their vector's immune system. Because CTL genes are important sugar-binding proteins for the innate immune response of C. elegans, the loss of some CTL genes of vector-transmitted PPNs might be responsible for their parallel adaptations to a mutualistic relationship with their vector. These results expand our understanding of the evolutionary benefits of vector-mediated transmission for the nematode and vector-nematode co-evolution.

A new bacteria-free strategy induced by MaGal2 facilitates pinewood nematode escape immune response from its vector beetle.

Symbiotic microbes play a crucial role in regulating parasite-host interactions; however, the role of bacterial associates in parasite-host interactions requires elucidation. In this study, we showed that, instead of introducing numerous symbiotic bacteria, dispersal of 4th-stage juvenile (JIV ) pinewood nematodes (PWNs), Bursaphelenchus xylophilus, only introduced few bacteria to its vector beetle, Monochamus alternatus (Ma). JIV showed weak binding ability to five dominant bacteria species isolated from the beetles' pupal chamber. This was especially the case for binding to the opportunistic pathogenic species Serratia marcescens; the nematodes' bacteria binding ability at this critical stage when it infiltrates Ma for dispersal was much weaker compared with Caenorhabditis elegans, Diplogasteroides asiaticus, and propagative-stage PWN. The associated bacterium S. marcescens, which was isolated from the beetles' pupal chambers, was unfavorable to Ma, because it caused a higher mortality rate upon injection into tracheae. In addition, S. marcescens in the tracheae caused more immune effector disorders compared with PWN alone. Ma_Galectin2 (MaGal2), a pattern-recognition receptor, was up-regulated following PWN loading. Recombinant MaGal2 protein formed aggregates with five dominant associated bacteria in vitro. Moreover, MaGal2 knockdown beetles had up-regulated prophenoloxidase gene expression, increased phenoloxidase activity, and decreased PWN loading. Our study revealed a previously unknown strategy for immune evasion of this plant pathogen inside its vector, and provides novel insights into the role of bacteria in parasite-host interactions.

Major ascaroside pheromone component asc-C5 influences reproductive plasticity among isolates of the invasive species pinewood nematode.

Pheromones are communication chemicals and regulatory signals used by animals and represent unique tools for organisms to mediate behaviors and make "decisions" to maximize their fitness. Phenotypic plasticity refers to the innate capacity of a species to tolerate a greater breadth of environmental conditions across which it adapts to improve its survival, reproduction, and fitness. The pinewood nematode, Bursaphelenchus xylophilus, an invasive nematode species, was accidentally introduced from North America into Japan, China, and Europe; however, few studies have investigated its pheromones and phenotypic plasticity as a natural model. Here, we demonstrated a novel phenomenon, in which nematodes under the condition of pheromone presence triggered increased reproduction in invasive strains (JP1, JP2, CN1, CN2, EU1, and EU2), while it simultaneously decreased reproduction in native strains (US1 and US2). The bidirectional effect on fecundity, mediated by presence/absence of pheromones, is henceforth termed pheromone-regulative reproductive plasticity (PRRP). We further found that synthetic ascaroside asc-C5 (ascr#9), the major pheromone component, plays a leading role in PRRP and identified 2 candidate receptor genes, Bxydaf-38 and Bxysrd-10, involved in perceiving asc-C5. These results suggest that plasticity of reproductive responses to pheromones in pinewood nematode may increase its fitness in novel environments following introduction. This opens up a new perspective for invasion biology and presents a novel strategy of invasion, suggesting that pheromones, in addition to their traditional roles in chemical signaling, can influence the reproductive phenotype among native and invasive isolates. In addition, this novel mechanism could broadly explain, through comparative studies of native and invasive populations of animals, a potential underlying factor behind of the success of other biological invasions.

VPTMdb: a viral posttranslational modification database.

In viruses, posttranslational modifications (PTMs) are essential for their life cycle. Recognizing viral PTMs is very important for a better understanding of the mechanism of viral infections and finding potential drug targets. However, few studies have investigated the roles of viral PTMs in virus-human interactions using comprehensive viral PTM datasets. To fill this gap, we developed the first comprehensive viral posttranslational modification database (VPTMdb) for collecting systematic information of PTMs in human viruses and infected host cells. The VPTMdb contains 1240 unique viral PTM sites with 8 modification types from 43 viruses (818 experimentally verified PTM sites manually extracted from 150 publications and 422 PTMs extracted from SwissProt) as well as 13 650 infected cells' PTMs extracted from seven global proteomics experiments in six human viruses. The investigation of viral PTM sequences motifs showed that most viral PTMs have the consensus motifs with human proteins in phosphorylation and five cellular kinase families phosphorylate more than 10 viral species. The analysis of protein disordered regions presented that more than 50% glycosylation sites of double-strand DNA viruses are in the disordered regions, whereas single-strand RNA and retroviruses prefer ordered regions. Domain-domain interaction analysis indicating potential roles of viral PTMs play in infections. The findings should make an important contribution to the field of virus-human interaction. Moreover, we created a novel sequence-based classifier named VPTMpre to help users predict viral protein phosphorylation sites. VPTMdb online web server (http://vptmdb.com:8787/VPTMdb/) was implemented for users to download viral PTM data and predict phosphorylation sites of interest.

Metabotropic glutamate receptor 5 regulates synaptic plasticity in a chronic migraine rat model through the PKC/NR2B signal.

BACKGROUND: The mechanism of chronic migraine (CM) is complex, central sensitization is considered as one of the pathological mechanism. Synaptic plasticity is the basis of central sensitization. Metabotropic glutamate receptor 5 (mGluR5) plays a vital role in the synaptic plasticity of the central nervous system. However, whether mGluR5 can promote the central sensitization by regulating synaptic plasticity in CM is unknown. METHODS: Male Wistar rats were used to establish a CM rat model, and the expression of mGluR5 mRNA and protein were detected by qRT-PCR and western blot. The allodynia was assessed by mechanical and thermal thresholds, and central sensitization was assessed by expression of the phosphorylation of cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) at Serine 133(pCREB-S133) and c-Fos. The synaptic-associated protein postsynaptic density protein 95 (PSD), synaptophysin (Syp), and synaptophysin-1(Syt-1), synaptic ultrastructure, and dendritic spines were detected to explore synaptic plasticity. The expression of PKC, total NR2B(tNR2B), and phosphorylation of NR2B at Tyr1472(pNR2B-Y1472) were detected by western blot. RESULTS: We found that the expression of mGluR5 was upregulated in CM rats. Downregulated the mGluR5 with MPEP alleviated the allodynia and reduced the expression of CGRP, pCREB-S133, c-Fos, PSD, Syp and Syt-1 and synaptic transmission. Moreover, the administration of MPEP inhibited the upregulation of PKC and pNR2B-Y1472. CONCLUSIONS: These results indicate that mGluR5 contributes to central sensitization by regulating synaptic plasticity in CM through the PKC/NR2B signal, which suggests that mGluR5 may be a potential therapeutic candidate for CM.