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Amoxicillin-clavulanate is the most common cause of idiosyncratic drug-induced liver injury (DILI). Drug-specific CD4+ T cells have been detected in patients with DILI, suggestive of an immune etiology. Furthermore, genetic associations including the human leucocyte antigen (HLA) DRB1*1501-DQB1*0602 haplotype influence susceptibility. see more Amoxicillin forms protein adducts that are postulated to activate T cells, by conjugating with lysine residues. However, a role for such adducts has not been described. This study aimed to (1) investigate whether amoxicillin-modified HLA-DRB1*1501-DQB1*0602 binding peptides selectively activate DILI patient T cells and (2) define the nature of the T-cell response with respective to antigen structure. Peptides carrying lysine residues for amoxicillin binding in positions (KP) 2-6 and anchors for the HLA-DRB1*1501-DQB1*0602 haplotype were designed. The amoxicillin-modified peptides were characterized by mass spectrometry prior to culturing with patient peripheral blood mononuclear cell. T-cell clones were then tested for specificity with amoxicillin, unmodified- and amoxicillin-modified peptides, and structural variants. Amoxicillin-modified KP-2 and KP-3 peptide-specific CD4+ clones proliferated and secreted interferon gamma (IFN-γ), interleukin (IL)-10, perforin and/or IL-17/IL-22 in a dose-dependent manner and displayed no cross-reactivity with amoxicillin, unmodified peptide or with positional derivatives. The T cells response was HLA class II restricted and the amoxicillin-modified peptides bound selectively to HLA-DRB1*1501 and/or DQB1*0602. To conclude, we show that amoxicillin-modified peptides bind to both components of the risk haplotype to stimulate DILI patient T cells and describe the importance of the position of nucleophilic lysine residue in the HLA binding peptide sequence.Both the mechanisms of monolignol transport and the transported form of monolignols in developing xylem of trees are unknown. We tested the hypothesis of an active, plasma membrane-localized transport of monolignol monomers, dimers, and/or glucosidic forms with membrane vesicles prepared from developing xylem and lignin-forming tissue-cultured cells of Norway spruce (Picea abies L. Karst.), as well as from control materials, comprising non-lignifying Norway spruce phloem and tobacco (Nicotiana tabacum L.) BY-2 cells. Xylem and BY-2 vesicles transported both coniferin and p-coumaryl alcohol glucoside, but inhibitor assays suggested that this transport was through the tonoplast. Membrane vesicles prepared from lignin-forming spruce cells showed coniferin transport, but the Km value for coniferin was much higher than those of xylem and BY-2 cells. Liquid chromatography-mass spectrometry analysis of membrane proteins isolated from spruce developing xylem, phloem, and lignin-forming cultured cells revealed multiple transporters. These were compared with a transporter gene set obtained by a correlation analysis with a selected set of spruce monolignol biosynthesis genes. Biochemical membrane vesicle assays showed no support for ABC-transporter-mediated monolignol transport but point to a role for secondary active transporters (such as MFS or MATE transporters). In contrast, proteomic and co-expression analyses suggested a role for ABC transporters and MFS transporters.Nitrogen (N) fertilizer maximizes the growth of oilseed rape (Brassica napus L.) by improving photosynthetic performance. Elucidating the dynamic relationship between fluorescence and plant N status could provide a non-destructive diagnosis of N status and the breeding of N-efficient cultivars. The aim of this study was to explore the impacts of different N treatments on photosynthesis at a spatial-temporal scale and to evaluate the performance of three fluorescence techniques for the diagnosis of N status. One-way ANOVA and linear discriminant analysis were applied to analyze fluorescence data acquired by a continuous excitation chlorophyll fluorimeter (OJIP transient analysis), pulse amplitude-modulated chlorophyll fluorescence (PAM-ChlF), and multicolor fluorescence (MCF) imaging. The results showed that the maximum quantum efficiency of PSII photochemistry (Fv/Fm) and performance index for photosynthesis (PIABS) of bottom leaves were sensitive to N status at the bolting stage, whereas the red fluorescence/far-red fluorescence ratio of top leaves was sensitive at the early seedling stage. Although the classification of N treatments by the three techniques achieved comparable accuracies, MCF imaging showed the best potential for early diagnosis of N status in field phenotyping because it had the highest sensitivity in the top leaves, at the early seedling stage. The findings of this study could facilitate research on N management and the breeding of N-efficient cultivars.Uridine diphosphate (UDP)-dependent glycosyltransferases catalyse the glycosylation of small molecules and play important roles in maintaining cell homeostasis and regulating plant development. Glycosyltransferases are widely distributed, but their detailed roles in regulating plant growth and development are largely unknown. In this study, we identified a UDP-glycosyltransferase, UGT85A53, from Camellia sinensis, the expression of which was strongly induced by various abiotic stress factors and its protein product was distributed in both the cytoplasm and nucleus. Ectopic overexpression of CsUGT85A53 in Arabidopsis resulted in an early-flowering phenotype under both long- and short-day conditions. The transcript accumulation of the flowering repressor genes FLC and ABI5, an activator of FLC in ABA-regulated flowering signaling, were both significantly decreased in transgenic Arabidopsis compared with wild-type plants. The decreased expression level of FLC might be associated with an increased level of DNA methylation that was observed in CsUGT85A53-overexpressing (OE) plants. Biochemical analyses showed that CsUGT85A53 could glucosylate ABA to form inactive ABA-glycoside in vitro and in planta. Overexpression of CsUGT85A53 in Arabidopsis resulted in a decreased concentration of free ABA and increased concentration of ABA-glucoside. The early-flowering phenotype in the CsUGT85A53-OE transgenic lines was restored by ABA application. Furthermore, CsUGT85A53-OE plants displayed an ABA-insensitive phenotype with higher germination rates compared with controls in the presence of low concentrations of exogenous ABA. Our findings are the first to identify a UGT in tea plants that catalyses ABA glucosylation and enhance flowering transition as a positive regulator.
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