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N-terminal acetylation (N -acetylation) is one of the most common protein modifications and plays crucial roles in viability and stress responses in animals and plants. However, very little is known about N -acetylation of viral proteins. Here, we identified the Thr residue at position 2 (Thr-2) in the βC1 protein encoded by the betasatellite of tomato yellow leaf curl China virus (TYLCCNB-βC1) as a novel N -acetylation site. Furthermore, the effects of TYLCCNB-βC1 N -acetylation on its function as a pathogenicity factor were determined via N -acetylation mutants in Nicotiana benthamiana plants. We found that N -acetylation of TYLCCNB-βC1 is critical for its self-interaction in the nucleus and viral pathogenesis, and that removal of N -acetylation of TYLCCNB-βC1 attenuated tomato yellow leaf curl China virus-induced symptoms and led to accelerated degradation of TYLCCNB-βC1 through the ubiquitin-proteasome system. Our data reveal a protective effect of N -acetylation of TYLCCNB-βC1 on its pathogenesis and demonstrate an antagonistic crosstalk between N -acetylation and ubiquitination in this geminiviral protein. • N-terminal acetylation (N -acetylation) is one of the most common protein modifications and plays crucial roles in viability and stress responses in animals and plants. • The Thr residue at position 2 (Thr-2) in the βC1 protein encoded by the betasatellite of tomato yellow leaf curl China virus (TYLCCNB-βC1) as a novel N -acetylation site. • N -acetylation of TYLCCNB-βC1 is critical for viral pathogenesis. [ABSTRACT FROM AUTHOR]

Vesicle-associated membrane protein (VAMP) belongs to the receptor protein on the membrane of the secretory transport vesicle and involves in host immune function. The intracellular pathogen Spiroplasma eriocheiris could cause Eriocheir sinensis tremor disease. In a previous study, it was found E. sinensis VAMP (EsVAMP) was differently expressed in S. eriocheiris infection by proteomics analysis. This study mainly aims at the function of EsVAMP in the process of the S. eriocheiris infection. The length of EsVAMP gene was 1681 bp, which contained a 395 bp open reading frame, 90 bp 5′-non-coding region (UTR) and 1277 bp 3′-UTR. The results of qPCR showed that EsVAMP was expressed highly in hemocytes and nerves, followed by gills, intestines and hepatopancreas, and lowly expressed in heart and muscles. EsVAMP in hemocytes was up-regulated after S. eriocheiris infection. After EsVAMP over-expression and S. eriocheiris infection, the RAW264.7 cell morphology and cell viability of the experiment group were significantly better than the control group. Meanwhile, the copy number of S. eriocheiris in the experiment group was significantly lower than that in the control group. After EsVAMP and pCMV-Cre-mCherry were ligated and transfected into RAW264.7 cells, it was found that EsVAMP and lysosome co-localized. Meanwhile, the phagocytosed inactivated S. eriocheiris number and phagocytosed efficiency in RAW264.7 cells were increased significantly. The interference experiment was carried out by synthesizing EsVAMP dsRNA to verify that the EsVAMP transcriptions were successfully suppressed. The S. eriocheiris copy number and the mortality of crab increased significantly after EsVAMP RNAi and S. eriocheiris infection. Meanwhile, the phagocytosed inactivated S. eriocheiris number and phagocytosed efficiency in hemocytes decreased significantly after EsVAMP RNAi and S. eriocheiris infection. These results showed that VAMP was involved in the cell phagocytosis to resist pathogen infection. • A vesicle-associated membrane protein, EsVAMP, was obtained from Eriocheir sinensis and related with Spiroplasma eriocheiris infection. • The cell morphology, cell viability was induced to resist S. eriocheiris infection after EsVAMP over-expression in RAW264.7 cells. • EsVAMP and lysosome was co-localized to increase the phagocytosis of RAW264.7 cells after EsVAMP over-expression in RAW264.7 cells. • The S. eriocheiris copy number and the mortality of crab was increased significantly and the hemocytes phagocytosis to inactivated S.eriocheiris decreased significantly after EsVAMP RNAi. [ABSTRACT FROM AUTHOR]

Fresh-cut produce yields mechanical damage and limits quality preservation. This research identifies the defence responses of fresh-cut broccoli (W (whole flower), F (floret), QF (quarterly-cut floret), FS (shredded floret)) during storage (0, 1, 2, 3 days) at room temperature. Consequently, changes during storage mainly occurred earlier (day 1), while fresh-cut changes primarily appeared at intense damage (FS). Uniquely accumulated quinic acid, 4-methoxyglucobrassicin, etc. were found in FS, while citric acid and glucose were exceptionally formed in W, F and QF. Importantly, a different glucosinolate breakdown system was identified in broccoli storage, encoded by myrosinases instead of TGGs in the pre-harvest system. Significant changes were also observed in the glucosinolates breakdown and de-novo biosynthesis as regulated by genes like PEN2/3, NIT, MYB122 and CYP81F1. Conclusively, candidate metabolites and genes identified in this study provide a reference for the precise preservation of cruciferous vegetables. • Unique organic acids were identified in different fresh-cut levels. • A new glucosinolate degrading system was discovered in postharvest broccoli. • Chlorogenic acid metabolism response immediately in fresh-cut broccoli. [ABSTRACT FROM AUTHOR]

Stress-activated protein kinase (SAPK) interacting protein 1 (SIN1) is an essential component of mTORC2. Previous studies have shown that SIN1 is a key regulator of Akt pathway which plays an important role in various pathological conditions including cancer. While its effects and mechanisms on the progression of NSCLC remain unknown. In this study, we report that SIN1 is able to promote the growth and migration of NSCLC cells both in vitro and in vivo. Overexpression of SIN1 promoted A549 and H1299 cells proliferation by both MTT and colony formation assays. Consistently, knockdown of SIN1 inhibited the proliferation of these cells. In transwell assay, overexpression of SIN1 increased the migration of A549 and H1299 cells, while SIN1 knockdown reduced their migration. In a tumor xenograft model, overexpression of SIN1 promoted tumor growth of A549 cells in vivo, while SIN1 knockdown suppresses the tumor growth. We also found a mechanistic link between SIN1 and H3K4me3, H3K4me3 is involved in SIN1 upregulation. Moreover, SIN1 can significantly promote the in vitro migration and invasion of NSCLC cells via induction epithelial mesenchymal transition (EMT) process, which subsequently leads to transcriptional downregulation of epithelial marker E-cadherin and upregulation of mesenchymal markers N-cadherin and Vimentin expression. Together, our results reveal that SIN1 plays an important role in NSCLC and SIN1 is a potential biomarker and a promising target in the treatment of NSCLC. [ABSTRACT FROM AUTHOR]

This study aimed to develop a fermentation process that allows the concomitant production of dextran (a texture-enhancing agent) and vitamin B12 (in situ fortification) for bread applications. Mixed fermentation of soya flour or rice bran using Propionibacterium freudenreichii DSM 20271 and Weissella confusa A16 with added sucrose resulted in substantial quantities of dextran (5.6–5.8% dry matter) and B12 (7.9–8.9 μg/100 g fresh weight), together with antifungal metabolites (e.g. acetic and propionic acids). In addition to an extended mould-free shelf life, the bread containing 50% (dough weight) fermented soya flour or rice bran not only contained adequate levels of B12 but also exhibited improved texture and sensory quality compared to the control, such as a higher loaf volume, a softer crumb, and a more cohesive and moister mouthfeel. The mixed fermentation reduced the beany or cooked-rice flavour but increased sour and cheesy flavours and aftertaste. Dextran produced during the fermentation process exhibited a masking effect on the beany and sour notes and aftertaste, consistently with reduced levels of green or grassy volatiles (e.g. hexanal, heptanal, (E , E)-2,4-decadienal, and 2-pentylfuran). Overall, the mixed fermentation method using P. freudenreichii DSM 20271 and W. confusa A16 showed potential for B12 fortification of bread products with high sensory quality. • Fermentation simultaneously produced dextran, vitamin B12, and propionate. • This led to improved texture quality and microbial shelf life of fortified bread. • Dextran exhibited a flavour-masking effect e.g. on beany and sour notes. • This was linked to less beany/green volatile compounds based on SPME-GC-MS. • Consuming 100 g of fortified bread would meet the recommended daily intake for B12. [ABSTRACT FROM AUTHOR]

The effects of limonene exposure on the growth of Microcystis aeruginosa and the release of toxic intracellular microcystin (MCY) were tested by evaluating the results obtained from the batch culture experiments with M. aeruginosa FACHB-905. The time series of cell as well as intracellular and extracellular MCY concentrations were evaluated during 5d of the incubation. After exposure to limonene, the number of cells gradually diminished; the net log cell reduction after 5d of the exposure was 3.0, 3.6, and 3.8log when the initial cell densities were set at 1.6×107, 1.1×106 and 4.1×105 cell/mL, respectively. Limonene was found to significantly influence the production and release of MCY. As the limonene exposure could inhibit the increase in the number of cells, the increase in the total MCY concentration in the medium was also inhibited. In the presence of limonene, the intracellular MCY was gradually released into the medium through a gradual reduction in the number of cells. The extracellular MCY concentration in the medium was significantly higher in the limonene-exposed samples than in the control samples, which confirmed that limonene cannot decompose the extracellular MCY. [Copyright &y& Elsevier]

Highlights: [•] USP22 was accumulated in nucleus. [•] We identified of a functional USP22 NLS. [•] The KRRK amino acid residues are indispensable in NLS. [•] The KRRK motif is conserved in USP22 homologues. [Copyright &y& Elsevier]

In order to prepare the kiwi wine with high nutritional characteristics and low alcohol content, the physicochemical properties, organic acids, monomer phenols, water-soluble vitamins and aroma of kiwi wine fermented by Saccharomyces cerevisiae and Wickerhamomyces anomalus in different inoculation method were analyzed. The results showed that the alcohol content of the three kiwi wines ranged from 5.3 to 5.5% (v/v). The lactic acid content of COF (inoculated with a mix of S. cerevisiae and W. anomalus) low-alcohol kiwi wine was 10.99 mg/mL. The quercetin and catechin contents of WSF (sequential inoculation with W. anomalus followed by S. cerevisiae) low-alcohol kiwi wine were significantly lower than those in the other kiwi wines. The aroma in COF and WSF low-alcohol kiwi wines were predominately from W. anomalus , while those in SWF (sequential inoculation with S. cerevisiae followed by W. anomalus) low-alcohol kiwi wine were predominately from S. cerevisiae. • Quinine and citric acid were the major organic acids in low-alcohol kiwi wine. • The major phenols of kiwi wines were protocatechuic acid, catechin and quercetin. • The aromas present in COF and WSF were predominately from W. anomalus. • The WSF low-alcohol kiwi wine had the highest sensory score. • The alcohol content of the three kiwi wines ranged from 5.3 to 5.5% (v/v). [ABSTRACT FROM AUTHOR]

To compensate the absence of viscoelastic gluten network, water-binding and viscosity-forming ingredients, hydrocolloids, are commonly used in gluten-free doughs. Currently, hydrocolloids are often E coded while many consumers prefer "clean-label" alternatives. For instance, microbial and cereal β-glucans are known to produce viscosity. In this study, the technological potential of these β-glucans as natural hydrocolloids was evaluated in gluten-free oat baking. Microbial β-glucan was produced by fermentation with P. claussenii using oat bran concentrate (OBC) as a raw material. Oat β-glucan was studied as various forms of OBC: as untreated OBC, pre-treated OBC suspension, and water-extractable fraction of OBC suspension (oat β-glucan extract). The baking properties after natural hydrocolloid additions were compared to a reference oat bread which was baked with common commercial hydrocolloids (psyllium, HPMC). The levels of hydrocolloids in the doughs were optimised by a test baking method. Among the hydrocolloids studied, the best bread properties were achieved with oat β-glucan extract as a hydrocolloid, as the breads resulted in similar specific volume and 37% lower staling rate compared to oat breads baked with commercial hydrocolloids. Thus, oat β-glucan extract provided a feasible and "clean-label" alternative for commercial hydrocolloids in gluten-free oat baking. • Oat β-glucan extract functioned well as a hydrocolloid in gluten-free oat baking. • Water-extraction increased β-glucan solubilisation and some structure was formed. • Additionally, removal of water-insoluble matter from oat bran concentrate was crucial. • With oat β-glucan extract, psyllium and HPMC could be replaced. • Oat β-glucan extract improved crumb properties and storage stability of oat bread. [ABSTRACT FROM AUTHOR]

Trichosanthes kirilowii has been mainly grown for use in traditional Chinese medicine. In this study, cucurbit mild mosaic virus (CuMMV) belonging to the genus Fabavirus was identified from T. kirilowii plants. CuMMV possesses a segmented, bipartite linear single-stranded RNA genome composed of RNA1 and RNA2. Sequence analysis showed that each genomic segment shares the highest sequence similarity with those of CuMMV isolated from pumpkin. A full-length infectious cDNA clone of CuMMV was further constructed and was found to induce typical symptoms in T. kirilowii , Cucumis sativus , C. melo , Citrullus lanatus , and Cucurbita pepo. The sap inoculum derived from the infectious cDNA clone of CuMMV could be mechanically transmitted and reproduce similar symptoms in the tested plants. This is the first report on the construction of a biologically active, full-length infectious cDNA clone of CuMMV, which will provide a useful tool in understanding CuMMV-encoded proteins and plant-CuMMV interactions. • The complete genome sequence of cucurbit mild mosaic virus (CuMMV) were cloned from Trichosanthes kirilowii. • An infectious cDNA clone of CuMMV was constructed and demonstrated to be highly infectious to T. kirilowii and other four cucurbitaceous plants. • CuMMV is mechanically transmissible to infect T. kirilowii and tested plants. [ABSTRACT FROM AUTHOR]

Excessive residual sulfur dioxide in black fungus is unfavorable to human health and has become an urgent problem to be solved. The purpose of our study is to evaluate the effect of non-thermal treatments on the degradation of residual sulfur dioxide in black fungus and their impacts on nutritional composition, and to explore whether non-thermal desulfurization changes the appearance characteristics (color and microstructure) of black fungus. After the synergistic effect of electrolyzed water and ultrasound (EWU), the degradation rate of sulfur dioxide was relatively ideal (degradation rate = 83.5%), and there was no loss of total sugar (TS) and total phenol (TP) in black fungus. EWU desulfurization caused minor damage to microstructure of black fungus, making its color darker. These results provide a reliable technical guarantee for EWU to remove sulfur dioxide from black fungus. • Non-thermal treatments effectively removed excessive sulfur dioxide in black fungus. • Electrolyzed water–ultrasonic synergy removed 83.5% of sulfur dioxide. • Electrolyzed water–ultrasonic synergy slightly damaged the appearance. • Electrolyzed water–ultrasonic synergy hardly affected main nutrients. [ABSTRACT FROM AUTHOR]

• The critical period of formaldehyde generation was identified and controlled. • RAA had the highest comprehensive score and high endogenous formaldehyde. • Total sugar and flavonoid are negatively correlated with endogenous formaldehyde. • GGT and C-S lyase together promote endogenous formaldehyde production. • Cys and Glu inhibited the production of endogenous formaldehyde. To maintain quality and reduce endogenous formaldehyde content in the edible fungus, Auricularia auricula , the critical period of formaldehyde generation was identified and controlled. In this study, different growth stages of A. auricula were used to assess the nutritional composition (polyphenol monomers, 5′-nucleotides, organic acids and water-soluble vitamins) and related enzyme activities; the correlation between quality and endogenous formaldehyde content was also analysed. The results showed that formaldehyde was positively correlated with soluble protein and melanin levels; it was negatively correlated with the total sugar and flavonoid contents. Among the four growth stages, ripe A. auricula (RAA) had the highest endogenous formaldehyde content. The activities of γ-glutamyl transpeptidase (GGT) and l -cysteine sulphoxide lyase (C-S lyase) were highest in RAA; they promoted endogenous formaldehyde production. RAA is a critical period for interventions to reduce endogenous formaldehyde contents. Experiments to distinguish the effects of 10 exogenous substances on GGT and C-S lyase indicated that cysteine (Cys) and glutamic acid (Glu) inhibited the production of endogenous formaldehyde by 17.13% and 14.89%, respectively. This study provides a new perspective for controlling safety during the growth of edible fungi and maintaining the quality of processed products. [ABSTRACT FROM AUTHOR]

Skp1-Cul1-F-box protein (SCF) ubiquitin E3 ligases play important roles in cancer development and serve as a promising therapeutic target in cancer therapy. Brusatol (Bru), a known Nrf2 inhibitor, holds promise for treating a wide range of tumors; however, the direct targets of Bru and its anticancer mode of action remain unclear. In our study, 793 Bru-binding candidate proteins were identified by using a biotin-brusatol conjugate (Bio-Bru) followed by streptavidin-affinity pull down-based mass spectrometry. We found that Bru can directly bind to Skp1 and disrupt the interactions of Skp1 with the F-box protein Skp2, leading to the inhibition of the Skp2-SCF E3 ligase. Bru inhibited both proliferation and migration via promoting the accumulation of the substrates p27 and E-cadherin; Skp1 overexpression attenuated while Skp1 knockdown enhanced these effects of Bru in non-small cell lung cancer (NSCLC) cells. Moreover, Bru binding to Skp1 also inhibited the β-TRCP-SCF E3 ligase. In both subcutaneous and orthotopic NSCLC xenografts, Bru significantly inhibited the growth and metastasis of NSCLC through targeting SCF complex and upregulating p27 and E-cadherin protein levels. These data demonstrate that Bru is a Skp1-targeting agent that may have therapeutic potentials in lung cancer. [Display omitted] • Biotin-brusatol conjugate and mass spectrometry were employed to identify that brusatol can target Skp1, an adaptor protein of SCF E3 ligase. • Brusatol binding to Skp1 leads to the inhibition of SCF E3 ligase such as Skp2-SCF E3 ligase, leading to the accumulation of E-cadherin and p27. • Brusatol at low concentrations (1–2 mg/kg) inhibits proliferation and metastasis of NSCLC xenografts via inhibiting SCF E3 ligase. [ABSTRACT FROM AUTHOR]

Macrophyte overgrowth in eutrophic lakes can hasten the decline of shallow water bodies, yet the impact of macrophyte deposition on sediment phosphorus (P) accumulation in the ice-on season remains unclear. Comparative analyses of P variations among 13 semi-connected sub-lakes in Wuliangsu Lake in China, a typical MDE lake, considered external flow and macrophyte decomposition as driving forces. Sediment P fractions and water total phosphorus (TP) were analyzed at 35 sampling points across three ice-on season stages, along with macrophyte TP content to assess debris contributions. Our findings reveal that phosphorus accumulation occurs during the ice-on season in the MDE lake, with an average TP content increase of 16 mg/kg. However, we observed a surprisingly small sediment nutrient accumulation ratio (ΔTP/ΔTN=0.006) compared to macrophyte nutrient levels before decomposition. Further analysis of the dominant species, Potamogeton pectinatus , indicates that a significant portion (55%) of macrophyte phosphorus is released before the ice-on season. This highlights the critical importance of timing macrophyte harvesting to precede the phosphorus leaching process, which has implications for lake management and ecosystem restoration efforts. Additionally, our research demonstrates similar transformations among different sediment fractions as previously reported. Macrophyte debris decomposition likely serves as the primary source of Residual P (Res-P) or TP accumulation. In addition, Ca-bound P (Ca–P) generally showed a decrease, which mainly caused by its transformation to Fe/Al-bound P (Fe/Al–P), Exchange-P (Ex-P), and sometimes to Res-P. However, we emphasize the significant impacts of flow dynamics on Ca–P transport and transformations. Its hydrodynamic action increases water dissolved oxygen, which accelerates the transformation of Ca–P to more easily released Fe/Al–P and Ex-P. Furthermore, hydrodynamic transport also leads to upstream Ca–P transport to downstream. This underscores the necessity of considering flow dynamics when estimating phosphorus variations and formulating phosphorus restoration strategies. [Display omitted] • Phosphorus (P) cycle in macrophyte-dominated eutrophic lakes remain understudied. • Comparatively analyzing P variations among 13 sub-lakes during ice-on season. • Residual P contributes to the P accumulation caused by macrophyte decomposition. • Fifty-five percent macrophyte P have been leached before the ice-on season. • Macrophyte harvesting remains necessary but should be done before P leaching. [ABSTRACT FROM AUTHOR]

Ralstonia solanacearum is a soil-borne pathogen that causes bacterial wilt worldwide. The virulence-attenuated mutants were able to combat the soil-borne plant diseases. In this study, we screened the virulence-attenuated mutant PRS-84-4-49 of Ralstonia solanacearum and demonstrated that this strain showed a significant biocontrol effect against patchouli bacterial wilt. Three putative virulence-attenuated mutants obtained in our previous preliminary screen were individually tested for their pathogenicity to patchouli plants. Mutant PRS-84-4-49 showed significantly less virulence to patchouli plants than the other investigated mutants. The virulence-attenuated mutant PRS-84-4-49 was then evaluated for its potential to control patchouli bacterial wilt. The results revealed that the biocontrol treatment significantly reduced disease severity compared with the inoculated control plants, their highest disease incidence were 33% and 63%, respectively, at 5 days post-inoculation. Mutant PRS-84-4-49 exhibited less motility and produced fewer biofilms than the wild-type strain. Therefore, our results demonstrate that virulence-attenuated mutant of Ralstonia solanacearum has potential as biological control agent capable of suppressing patchouli bacterial wilt. • This work investigated the role of virulence-attenuated mutant PRS-84-4-49 of Ralstonia solanacearum in suppressing patchouli bacterial wilt. • The Ralstonia solanacearum mutant exhibited less motility and produced fewer biofilms than the wild-type strain. [ABSTRACT FROM AUTHOR]

EEF1A2 encodes protein elongation factor 1-alpha 2, which is involved in Guanosine triphosphate (GTP)-dependent binding of aminoacyl-transfer RNA (tRNA) to the A-site of ribosomes during protein biosynthesis and is highly expressed in the central nervous system. De novo mutations in EEF1A2 have been identified in patients with extensive neurological deficits, including intractable epilepsy, globe developmental delay, and severe intellectual disability. However, the mechanism underlying phenotype variation is unknown. Using next-generation sequencing, we identified a novel and a recurrent de novo mutation, c.294C > A; p.(Phe98Leu) and c.208G > A; p.(Gly70Ser), in patients with Lennox–Gastaut syndrome. The further systematic analysis revealed that all EEF1A2 mutations were associated with epilepsy and intellectual disability, suggesting its critical role in neurodevelopment. Missense mutations with severe molecular alteration in the t-RNA binding sites or GTP hydrolysis domain were associated with early-onset severe epilepsy, indicating that the clinical expression was potentially determined by the location of mutations and alteration of molecular effects. This study highlights the potential genotype–phenotype relationship in EEF1A2 and facilitates the evaluation of the pathogenicity of EEF1A2 mutations in clinical practice. • We identified two de novo mutations in EEF1A2 in patients with LGS. • c.294C > A was novel and c.208G > A was a recurrent de novo mutation. • All EEF1A2 mutations were associated with epilepsy and intellectual disability. • Mutations in key functional domains were associated with early-onset severe epilepsy. • Systematic analysis highlights the potential genotype–phenotype relationship. [ABSTRACT FROM AUTHOR]