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V-domain Ig suppressor of T cell activation (VISTA), a novel immune checkpoint regulatory molecule, suppresses T cell mediated immune responses. The aim of the present study was to profile the immunological expression, clinical significance and correlation of VISTA in human oral squamous cell carcinoma (OSCC). Human tissue microarrays, containing 165 primary OSCCs, 48 oral epithelial dysplasias and 43 normal oral mucosae, were applied to investigate the expression levels of VISTA, CD8, cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), programmed death ligand 1 (PD-L1), PI3Kα p110, IL13Rα2, phospho-STAT3 at tyrosine 705 (p-STAT3) and myeloid-derived suppressor cell (MDSC) markers (CD11b and CD33) by immunohistochemistry and digital pathology analysis. The results demonstrated that the protein level of VISTA was significantly higher in human OSCC specimens, and that VISTA expression in primary OSCCs was correlated with lymph node status. VISTA expression did not serve as an independent predictor for poor prognosis, while patient subgroup with VISTA high and CD8 low expression (22/165) had significantly poorer overall survival compared with other subgroups based on the multivariate and Cox hazard analyses among the primary OSCC patients in the present cohort. Additionally, the expression of VISTA was significantly correlated with PD-L1, CTLA-4, IL13Rα2, PI3K, p-STAT3, CD11b and CD33 according to Pearson's correlation coefficient test. Taken together, the results indicated that the VISTA high and CD8 low group, as an immunosuppressive subgroup, might be associated with a poor prognosis in primary OSCC. These findings indicated that VISTA might be a potential immunotherapeutic target in OSCC treatment. [ABSTRACT FROM AUTHOR]

Checkpoint blockade therapy utilizing monoclonal antibodies to reactivate T cells and recover their antitumor activity makes an epoch in cancer immunotherapy. The role of B7-H4, a novel negative immune checkpoint, in oral squamous cell carcinoma (OSCC) has still not been elucidated. In this study, tissue samples from human OSCC, which contains 165 primary OSCC, 48 oral epithelial dysplasia and 43 normal oral mucosa specimens, and Tgfbr1/ Pten 2cKO mice OSCC model were stained with B7-H4 antibody to analyze the correlations between B7-H4 expression and clinicopathological characteristics. Kaplan-Meier analysis was used to compare the survival of patients with high B7-H4 expression and patients with low B7-H4 expression. We found B7-H4 is highly expressed in human OSCC tissue, and the B7-H4 expression level was associated with the clinicopathological parameters containing pathological grade and lymph node status. Moreover, we confirmed that B7-H4 was overexpressed in Tgfbr1/ Pten 2cKO mice OSCC model. Our data also indicated that patients with high B7-H4 expression had poor overall survival compared with those with low B7-H4 expression. Furthermore, this study demonstrated that B7-H4 was positively associated with PD-L1, CD11b, CD33, PI3Kα p110, and p-S6 (S235/236). Taken together, these findings suggest B7-H4 is a potential target in the treatment of OSCC. [ABSTRACT FROM AUTHOR]

Abstract: Cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) is considered to be a key enzyme in lignin biosynthesis, but little was known about CADs in tea plants (Camellia sinensis). A full-length cDNA sequence (CsCAD2) was isolated by suppressive subtractive hybridization (SSH) in Ectropis oblique feeding-induced tea plants, and another two full-length cDNA sequences (CsCAD1 and CsCAD3) were obtained from a transcriptome obtained by deep sequencing. However, they showed only 20–54% identities. Phylogenetic analysis revealed that they belonged to three different families. DNA gel blotting analysis revealed that two copies of CsCAD1 and CsCAD2 genes existed in tea genome, but CsCAD3 likely had only one copy. Recombinant proteins of these CsCADs were produced in Escherichia coli. The activity of purified recombinant CsCAD2 protein was up to 0.43μmolmin−1 mg−1. However, the other two recombinant proteins had lower activities, probably due to incomplete refolding. qRT-PCR analysis indicated that while CsCAD3 was strongly up-regulated in tea plants after E. oblique attack and mechanical damage, CsCAD1 and CsCAD2 showed only moderate or no changes in transcript levels. Treatment of defence-related hormones methyl jasmonate (MeJA) and salicylic acid (SA) elevated the expression of CsCAD1 and CsCAD2, but decreased the transcript abundance of CsCAD3. The transcript levels of CsCAD2 did not change after applying abscisic acid (ABA), whereas CsCAD1 and CsCAD3 were induced. These results suggested that these three CsCAD genes in tea plants may play a role in defense against insects and pathogens and adaptation to abiotic stresses and these genes likely have divergant functions. [Copyright &y& Elsevier]

Pyridine compounds, including nicotinic acid and nicotinamide, are key metabolites of both the salvage pathway for NAD and the biosynthesis of related secondary compounds. We examined the in situ metabolic fate of [carbonyl-C]nicotinamide, [2-C]nicotinic acid and [carboxyl-C]nicotinic acid riboside in tissue segments of tea ( Camellia sinensis) plants, and determined the activity of enzymes involved in pyridine metabolism in protein extracts from young tea leaves. Exogenously supplied C-labelled nicotinamide was readily converted to nicotinic acid, and some nicotinic acid was salvaged to nicotinic acid mononucleotide and then utilized for the synthesis of NAD and NADP. The nicotinic acid riboside salvage pathway discovered recently in mungbean cotyledons is also operative in tea leaves. Nicotinic acid was converted to nicotinic acid N-glucoside, but not to trigonelline ( N-methylnicotinic acid), in any part of tea seedlings. Active catabolism of nicotinic acid was observed in tea leaves. The fate of [2-C]nicotinic acid indicates that glutaric acid is a major catabolite of nicotinic acid; it was further metabolised, and carbon atoms were finally released as CO. The catabolic pathway observed in tea leaves appears to start with the nicotinic acid N-glucoside formation; this pathway differs from catabolic pathways observed in microorganisms. Profiles of pyridine metabolism in tea plants are discussed. [ABSTRACT FROM AUTHOR]

Abstract: Theanine synthetase (TS) is an enzyme involved in theanine biosynthesis in tea plants. Recent studies have revealed that theanine biosynthesis, derived from nitrogen metabolism in tea (Camellia sinensis L.) plants, could be influenced by salt treatment. We have characterized CsTS at the molecular and biochemical level. The expression pattern of CsTS protein was examined by western blot using a self-prepared polyclonal antibody with high specificity and sensitivity. The effect of salt treatment on the levels of theanine synthesis was investigated in this study. Levels of theanine and the total free amino acids were gradually increased in shoots, and reached the maximum on the 8th day after treatment (DAT). The immunoblotting analysis suggested the accumulation of CsTS protein had increased gently up to 8 DAT, and subsequently declined, both in roots and shoots, which is one of the main evidences that resulted in the variation of theanine concentration under salt treatment. Together, these data revealed that theanine synthesis takes place both in root and shoot and CsTS accumulation is positively affected by salt treatment. [Copyright &y& Elsevier]

To determine the metabolic profiles of purine nucleotides and related compounds in leaves and roots of tea (Camellia sinensis), we studied the in situ metabolic fate of 10 different 14C-labeled precursors in segments from tea seedlings. The activities of key enzymes in tea leaf extracts were also investigated. The rates of uptake of purine precursors were greater in leaf segments than in root segments. Adenine and adenosine were taken up more rapidly than other purine bases and nucleosides. Xanthosine was slowest. Some adenosine, guanosine and inosine was converted to nucleotides by adenosine kinase and inosine/guanosine kinase, but these compounds were easily hydrolyzed, and adenine, guanine and hypoxanthine were generated. These purine bases were salvaged by adenine phosphoribosyltransferase and hypoxanthine/guanine phosphoribosyltransferase. Salvage activity of adenine and adenosine was high, and they were converted exclusively to nucleotides. Inosine and hypoxanthine were salvaged to a lesser extent. In situ 14C-tracer experiments revealed that xanthosine and xanthine were not salvaged, although xanthine phosphoribosyltransferase activity was found in tea extracts. Only some deoxyadenosine and deoxyguanosine was salvaged and utilized for DNA synthesis. However, most of these deoxynucleosides were hydrolyzed to adenine and guanine and then utilized for RNA synthesis. Purine alkaloid biosynthesis in leaves is much greater than in roots. In situ experiments indicate that adenosine, adenine, guanosine, guanine and inosine are better precursors than xanthosine, which is a direct precursor of a major pathway of caffeine biosynthesis. Based on these results, possible routes of purine metabolism are discussed. [ABSTRACT FROM AUTHOR]

Abstract: The distribution of phenolic compounds in young and developing leaves, stems, main and lateral roots and cotyledons of 8-week-old tea (Camellia sinensis) seedlings was investigated using HPLC-MS2. Fourteen compounds, flavan-3-ols, chlorogenic acids, and kaempferol-O-glycosides, were identified on the basis of their retention time, absorbance spectrum, and MS fragmentation pattern. The major phenolics were (−)-epigallocatechin-3-O-gallate and (−)-epicatechin-3-O-gallate, located principally in the green parts of the seedlings. Considerable amounts of radioactivity from [ring-14C]phenylalanine were incorporated in (−)-epicatechin, (−)-epigallocatechin, (−)-epicatechin-3-O-gallate and (−)-epigallocatechin-3-O-gallate, by tissues of young and developing leaves and stems. Expression of genes encoding enzymes involved in flavan-3-ol biosynthesis, CHS, CHI, F3H, F3′5′H, DFR, ANS, ANR and LAR was investigated. Transcripts of all genes, except LAR, were more abundant in leaves and stems than in roots and cotyledons. No significant difference was found in the amount of transcript of LAR. These findings indicate that in tea seedlings flavan-3-ols are produced by a naringenin-chalcone→naringenin→dihydrokaempferol pathway. Dihydrokaempferol is a branch point in the synthesis of (−)-epigallocatechin-3-O-gallate and other flavan-3-ols which can be formed by routes beginning with either a flavonoid 3′-hydroxylase mediated conversion of the flavonol to dihydroquercetin or a flavonoid 3′,5′-hydroxylase-catalysed conversion to dihydromyricetin with subsequent steps involving sequential reactions catalysed by dihydroflavanol 4-reductase, anthocyanidin synthase, anthocyanidin reductase and flavan-3-ol gallate synthase. [Copyright &y& Elsevier]

Abstract: ATP sulfurylase, the first enzyme in the sulfate assimilation pathway of plants, catalyzes the formation of adenosine phosphosulfate from ATP and sulfate. Here we report the cloning of two cDNAs encoding ATP sulfurylase (APS1 and APS2) from Camellia sinensis. They were isolated by RT-PCR and RACE-PCR reactions. The expression of APS1 and APS2 are correlated with the presence of ATP sulfurylase enzyme activity in cell extracts. APS1 is a 1415-bp cDNA with an open reading frame predicted to encode a 360-amino acid, 40.5kD protein; APS2 is a 1706-bp cDNA with an open reading frame to encode a 465-amino acid, 51.8kD protein. The predicted amino acid sequences of APS1 and APS2 have high similarity to ATP sulfurylases of Medicago truncatula and Solanum tuberosum, with 86% and 84% identity respectively. However, they share only 59.6% identity with each other. The enzyme extracts prepared from recombinant Escherichia coli containing Camellia sinensis APS genes had significant enzyme activity. [Copyright &y& Elsevier]

The phenotype of albino tea plants (ATPs) is significantly influenced by temperature regimes and light conditions, which alter certain components of the tea leaves leading to corresponding phenotypic changes. However, the regulatory mechanism of temperature-dependent changes in photosynthetic pigment contents and the resultant leaf colors remain unclear. Here, we examined the chloroplast microstructure, shoot phenotype, photosynthetic pigment content, and the expression of pigment synthesis-related genes in three tea genotypes with different leaf colors under different temperature conditions. The electron microscopy results revealed that all varieties experienced the most severe chloroplast damage at 15 °C, particularly in albino cultivar Baiye 1 (BY), where chloroplast basal lamellae were loosely arranged, and some chloroplasts were even empty. In contrast, the chloroplast basal lamellae at 35 °C and 25 °C were neatly arranged and well-developed, outperforming those observed at 20 °C and 15 °C. Chlorophyll and carotenoid measurements revealed a significant reduction in chlorophyll content under low temperature treatment, peaking at ambient temperature followed by high temperatures. Interestingly, BY showed remarkable tolerance to high temperatures, maintaining relatively high chlorophyll content, indicating its sensitivity primarily to low temperatures. Furthermore, the trends in gene expression related to chlorophyll and carotenoid metabolism were largely consistent with the pigment content. Correlation analysis identified key genes responsible for temperature-induced changes in these pigments, suggesting that changes in their expression likely contribute to temperature-dependent leaf color variations. [ABSTRACT FROM AUTHOR]

Objectives: To examine the predictive value of dual-layer spectral detector CT (DLCT) for spread through air spaces (STAS) in clinical lung adenocarcinoma. Methods: A total of 225 lung adenocarcinoma cases were retrospectively reviewed for demographic, clinical, pathological, traditional CT, and spectral parameters. Multivariable logistic regression analysis was carried out based on three logistic models, including a model using traditional CT features (traditional model), a model using spectral parameters (spectral model), and an integrated model combining traditional CT and spectral parameters (integrated model). Receiver operating characteristic (ROC) curve analysis and decision curve analysis (DCA) were performed to assess these models. Results: Univariable analysis showed significant differences between the STAS and non-STAS groups in traditional CT features, including nodule density (p < 0.001), pleural indentation types (p = 0.006), air-bronchogram sign (p = 0.031), the presence of spiculation (p < 0.001), long-axis diameter of the entire nodule (LD) (p < 0.001), and consolidation/tumor ratio (CTR) (p < 0.001). Multivariable analysis revealed that LD > 20 mm (odds ratio [OR] = 2.271, p = 0.025) and CTR (OR = 24.208, p < 0.001) were independent predictors in the traditional model, while electronic density (ED) in the venous phase was an independent predictor in the spectral (OR = 1.062, p < 0.001) and integrated (OR = 1.055, p < 0.001) models. The area under the curve (AUC) for the integrated model (0.84) was the highest (spectral model, 0.83; traditional model, 0.80), and the difference between the integrated and traditional models was statistically significant (p = 0.015). DCA showed that the integrated model had superior clinical value versus the traditional model. Conclusions: DLCT has added value for STAS prediction in lung adenocarcinoma. Clinical relevance statement: Spectral CT has added value for spread through air spaces prediction in lung adenocarcinoma so may impact treatment planning in the future. Key Points: • Electronic density may be a potential spectral index for predicting spread through air spaces in lung adenocarcinoma. • A combination of spectral and traditional CT features enhances the performance of traditional CT for predicting spread through air spaces. [ABSTRACT FROM AUTHOR]

Abstract: The theanine content of the leaves of 27 species or varieties of Theaceae plants was investigated. Theanine was present in 21 species or varieties, but in much lower amounts (<0.2 μmol/g fresh weight) than the quantity detected in Camellia sinensis var. sinensis. The major free amino acids in leaves of four species belonging to the genera Schima and Eurya, were glutamic acid, aspartic acid, glutamine, asparagine, alanine and proline and content of these amino acids is similar to or higher than theanine. Accumulation of free amino acids in these plants was generally lower than in C. sinensis var. sinensis. The biosynthetic activity of theanine, assessed by the incorporation of radioactivity from [14C]ethylamine, was detected in seedlings of two species of Schima. The theanine biosynthetic activity in roots was higher than that of leaves. [Copyright &y& Elsevier]

Tea is a healthy beverage made from the leaves of the tea plant [Camellia sinensis (L.) O. Kuntze]. The tea plant is a perennial evergreen plant that is widely distributed in tropical and subtropical regions. PR proteins (pathogenesis-related proteins, PRs) are defense-related proteins induced under pathogenesis-related conditions. Currently, there are a few studies on PRs in plants. Tea leaf color mutants have been of wide interest to scientists as special materials. A tea plant displaying variegated spontaneous albinism found in a local tea plantation in Huangshan City, Anhui Province, China, was used as the material sample in this study. An analysis of preexisting transcriptomic and proteomic data revealed the significant upregulation of three classes of PRs. In order to investigate the correlation between PRs and variegated tea plants, a series of studies were conducted on PR genes. The results revealed the identification of 17 CsPR1, 3 CsPR4 and 31 CsPR5 genes in tea plants using bioinformatics methods, and their structures and promoter sequences. The expression of three PR1, two PR4 and one PR5 genes was determined to be induced in stress treatment experiments involving mechanical damage, insect bites, low temperature treatment, and fungal infections. Additionally, the ribonuclease activity of CsPR4 was successfully verified. This is the first study to report the ribonuclease activity of CsPR4 in tea plants. The results can serve as a reference for future studies on PRs in tea plants, offering new insights into information on albinism in tea plants. [ABSTRACT FROM AUTHOR]

Immunotherapy with immune checkpoint molecule-specific monoclonal antibody have obtained encouraging results from preclinical studies and clinical trials, which promoted us to explore whether this kind of immunotherapy could be applicable to head and neck squamous cell carcinoma (HNSCC). Lymphocyte activation gene-3 (LAG-3) is an immune checkpoint control protein that negatively regulates T cells and immune response. Here, using the human tissue samples, we report these findings that LAG-3 is overexpressed on tumor-infiltrating lymphocytes (TILs;p< 0.001) and its overexpression correlates with the high pathological grades, lager tumor size and positive lymph node status in human primary HNSCC. Survival analysis identifies LAG-3 as a prognostic factor independent of tumor size and pathological grades for primary HNSCC patients with negative lymph node status (p= 0.014). Study in immunocompetent genetically defined HNSCC mouse model reports that LAG-3 is upregulated on CD4+T cells, CD8+T cells and CD4+Foxp3+regulatory T cells (Tregs).In vivostudy, administration of LAG-3-specific antibody retards tumor growth in a way associated with enhanced systemic antitumor response by potentiating the antitumor response of CD8+T cells and decreasing the population of immunosuppressive cells. Taken together, our results offer a preclinical proof supporting the immunomodulatory effects of LAG-3 and suggest a potential therapeutic target of immunotherapy for HNSCC. [ABSTRACT FROM AUTHOR]

Integrin β4 (ITGB4) is a member of the integrin family, which plays a crucial role in mediating cell adhesion to the extracellular matrix. Recent studies have demonstrated that ITGB4 is involved in tumorigenesis and metastasis during the development of cancer. However, the role of ITGB4 in oral squamous cell carcinoma (OSCC) remains unclear. A Multiplex immunohistochemistry (OPAL™, mIHC) assay was employed to stain ITGB4, ALDH1, PD-L1, cytokeratin (CK), CD8 and PD-1 in a human OSCC tissue microarray, containing 26 normal oral epithelium samples, 21 oral epithelium dysplasia samples and 76 OSCC samples. The expression pattern and clinicopathological characteristics of ITGB4 were analyzed and compared with those of PD-1, PD-L1, ALDH1 and CD8. The correlation between subgroups of tumor cells, including ITGB4+PD-L1+ and ITGB4+ALDH1+, and subgroups of T cells, including CD8+ and CD8+PD-1+, was evaluated using two-tailed Pearson's statistics. A Kaplan–Meier curve was built, and a log-rank test was performed to analyze the survival rate of different subgroups. The mIHC staining results show that ITGB4 was mostly expressed in the tumor cells, with a significant increase in the OSCC specimens compared with normal oral epithelium and oral epithelium dysplasia. The paired analysis, conducted between the OSCC tumor tissue and normal paracancer mucosa, confirmed the results. The study further revealed that ITGB4+PD-L1+ cancer cells, but not ITGB4+ALDH1+ cancer cells, were significantly associated with the infiltration of CD8+ T cells (positivity p = 0.005, positive number p = 0.03). Additionally, ITGB4+PD-L1+ tumor cells were positively correlated with CD8+PD-1+ T cells (positivity p = 0.02, positive number p = 0.03). Most intriguingly, the subgroup of ITGB4/PD-L1high with CD8/PD-1high displayed the best prognosis compared with the other considered subgroups. The results show that the expression of ITGB4 was increased in OSCC compared with normal oral mucosa. Furthermore, a specific subgroup with high levels of expression of ITGB4/PD-L1 and CD8/PD-1 was found to have a relatively better prognosis compared with the other subgroups. Ultimately, this study sheds light on the potential role of ITGB4 in OSCC and provides a basis for further investigation. [ABSTRACT FROM AUTHOR]

ABSTRACT Objective The specific purposes of this study were (1) to undertake a thorough systematic review and meta-analysis based only on randomized clinical trials (RCTs) to compare the rates of survival and complications of short implants to those of long implants; (2) to compare the surgical time and cost of short implants to those of long implants. Methods RCTs were identified from the major electronic databases (MEDLINE, Embase and Cochrane Library) using the keywords 'dental implant,' 'short implant' and 'atrophic maxilla,' and a quantitative meta-analysis was conducted. The survival rate of implants and complications were the primary outcome measures, and other parameters assessed included costs and surgical time. Results Seven RCTs that met the inclusion criteria included 554 implants (265 implants in the short implant group). There was no significant difference in survival rate between two groups (RR: 1.00; 95% CI: [0.97, 1.03]; p = .96; seven trials, 554 participants). Compared with long implant group, the short implant group had a lower complications and the effect measure was significant (RR: 0.58; 95% CI: [0.37, 0.90]; p = .02; seven trials, 554 participants). Conclusion This systematic review showed that no difference between the survival rates of short implants (5-8 mm) and long implants (>8 mm); complications in short implants are lower than that in long implants. However, further studies are required to substantiate our findings. [ABSTRACT FROM AUTHOR]

In this study, quantitative taste evaluation based on sense and electronic tongue (ET) combined with chemical composition analysis reveals the taste quality difference of Keemun black tea (KBT) with natural withering, sun withering and warm-air withering treatments. The results showed that there were significant differences between the taste of the three finished teas, both in sensory and ET evaluation, indicating a stronger bitterness and astringency in the sun withering and a weaker bitterness and astringency in the warm-air withering. The quantitative results of LC-MS and HPLC on the taste compounds indicated that the total amount of catechins in sun withering was 20.85 mg/g, which was 1.90 mg/g and 5.06 mg/g higher than that in natural withering and warm air withering, respectively. This may be because sunlight promotes the conversion of theanine to catechins. The correlation results indicated that there was a quantitative and effective relationship between taste and tasting compounds and that there was a synergistic effect (R ≥ 0.8) between sweet and astringent compounds. This study emphasised a comparative evaluation in methods and in the taste effect of different withering methods, especially providing a theoretical basis for the quality control and flavour improvement of KBT. • Withering methods played a vital role in taste quality of KBT. • Sun withering improved the catechins content. • Warm-air withering enhanced the umami and sweetness of tea infusion. • Catechins and flavonoid glycosides were the main astringency contributors to KBT. [ABSTRACT FROM AUTHOR]

Unlike that of other crops, the growth of tea plants can be promoted by aluminum, but its regulation mechanism remains unclear. Some endophytes can also promote growth of plant hosts. In this paper, tea roots treated with aluminum were used to study the growth-promoting traits and aluminum tolerance of endophytes. Meta-16S rDNA analysis revealed that Burkholderia was enriched in tea roots after aluminum treatment, and it was the dominant strain for hydroponic tea roots and field tea roots. Actinomycetes constituted the dominant strains in hydroponic tea seedlings treated with aluminum. Sixteen endophytic bacteria, including 12 strains of Firmicutes , 2 strains of Proteobacteria and 2 strains of Actinomycetes , were isolated and identified from hydroponic tea roots treated with different aluminum concentrations. Growth-promoting activity analysis showed that the isolated endophytic bacteria all had more than one plant growth-promoting trait. Among them, B4 (Bacillus nealsonii), B8 (Brevibacterium frigoritolerans) and A2 (Nocardia nova) bacteria each had three growth-promoting traits. Aluminum tolerance ability analysis indicated that endophyte A1 (Leifsonia shinshuensis) had the strongest aluminum tolerance ability, up to 200 mg l−1 aluminum. Plant–bacteria interactions showed that endophytes A1, A2 and B4 and their synthetic community all had a growth-promoting effect on the growth of wheat lateral roots. Moreover, endophytes A1 and B4 alleviated aluminum stress in wheat. Endophyte A1 also promoted the growth of tea cuttings, especially lateral roots, with/without aluminum. Taken together, aluminum enhanced the distribution of aluminum-tolerant and growth-promoting bacteria, thereby promoting the growth of tea roots. This study provides a new aspect for research on the mechanism by which aluminum promotes tea plant growth. [ABSTRACT FROM AUTHOR]

Abstract: The metabolic fate of [carbonyl-14C]nicotinamide was surveyed in leaf disks of seven mangrove species, Bruguiera gymnorrhiza, Rhizophora stylosa, Kandelia obovata, Sonneratia alba, Pemphis acidula, Lumnitzera racemosa and Avicennia marina, with and without 250mM NaCl. Uptake of [14C]nicotinamide by leaf disks was stimulated by 250mM NaCl in K. candel, R. stylosa, A. marina and L. racemosa. [Carbonyl-14C]nicotinamide was converted to nicotinic acid and was utilised for the synthesis of nucleotides and nicotinic acid conjugates. Formation of nicotinic acid by the deaminase reaction was rapid; there was little accumulation of nicotinamide in the disks 3h after administration. Radioactivity from [carbonyl-14C]nicotinamide was incorporated into pyridine nucleotides (mainly NAD and NADP) in all mangrove leaves, and the rates varied from 2% (in L. racemosa) to 15% (S. alba) of the total radioactivity taken up. NaCl generally reduced nicotinic acid salvage for NAD and NADP. In all mangrove leaf disks, the most heavily labelled compounds (up to 70% of total radioactivity) were trigonelline (N-methylnicotinic acid) and/or nicotinic acid N-glucoside. Trigonelline was formed in all mangrove plants, but N-glucoside synthesis was found only in leaves of A. marina and K. obovata. In A. marina, incorporation of radioactivity into N-glucoside (51%) was much greater than incorporation into trigonelline (2%). In general, NaCl stimulates the synthesis of these pyridine conjugates. The rate of decarboxylation of nicotinic acid in roots of A. marina seedlings was much greater than for the corresponding reaction observed in leaves. [Copyright &y& Elsevier]

Crush–tear–curl (CTC) black tea is a popular beverage, owing to its unique taste characteristics and health benefits. However, differences in the taste quality and chemical profiles of CTC black tea from different geographical regions remain unclear. In this study, 28 CTC black tea samples were collected from six geographical regions and analyzed using electronic tongue and ultrahigh performance liquid chromatography‐Orbitrap‐mass spectroscopy. The e‐tongue analysis indicated that each region's CTC black tea has its own relatively prominent taste characteristics: Sri Lanka (more umami and astringent), North India (more umami), China (more sweetness and astringent), South India (moderate umami and sweetness), and Kenya (moderate umami and astringent). Based on multivariate statistical analysis, 78 metabolites were tentatively identified and used as potential markers for CTC black tea of different origins, mainly including amino acids, flavone/flavonol glycosides, and pigments. Different metabolites, which contributed to the taste characteristics of CTC black tea, were clarified by partial least squares regression correlation analysis. Our findings may serve as useful references for future studies on origin traceability and quality characteristic determination of CTC black teas. Practical Application: This study provides useful references for future studies on the origin traceability and taste characteristic determination of CTC black teas from different geographical regions. [ABSTRACT FROM AUTHOR]

Drying plays a key role in the formation of the final aroma of black tea. However, the contribution to its aroma remains unclear. Herein, headspace solid-phase microextraction (HP-SPME) and solvent-assisted flavor evaporation (SAFE) coupled with GC-MS were used to detect the volatiles of tea samples (before or after drying) from three cultivars. We found that the drying step caused increase in sweet and fruity aromas and decrease in mint-like and green aromas. 89 Differential volatiles were identified with variable importance in projection (VIP) over 1. Furthermore, 25 decreased aroma-active volatiles were obtained with relative odor activity values (rOAVs) over 1, showing that green, flowery, fruity, sweet, and mint-like decreased significantly by 62.27%, 64.88%, 67.56%, 58.26%, and 69.72% after drying step. (Z)-3-Hexen-1-ol (rOAV = 36.93), linalool (rOAV = 301.56) and methyl salicylate (rOAV = 5.50) were the odorants for the green, flowery, fruity, and mint-like odor. Moreover, other 3 increased aroma-active volatiles showed that flowery, fruity, and sweet volatiles increased significantly after drying step. β -Damascenone (rOAV = 43.49) was the main odorant for fruity and sweet aroma. The 28 differential volatiles were associated with carotenoid degradation, lipid degradation, and glycoside hydrolysis. This study reveals the contribution of drying to the aroma formation of black tea. • The drying step enhanced the sweet and fruity aromas of black tea. • The drying step weakened the green and mint-like aromas of black tea. • β -Damascenone was the main odorant for fruity and sweet aroma enhancement. • Increased aroma-active volatiles were mainly related to the carotenoid degradation. [ABSTRACT FROM AUTHOR]

[Display omitted] • Over-fired drying can significantly improve the quality of LAGP tea. • LC–MS, HPLC and GC–MS revealed the taste and aroma compounds due to TSD and PLD. • A total of 5 compounds related to taste were selected by DOT > 1. • Concentrations of volatiles were 45.46% higher after PLD treatment. Lu'an Guapian (LAGP) tea, a representative Chinese roasted green tea, undergoes significant changes in taste and aroma during over-fired drying. However, limited studies have been conducted on these effects. This study employed metabolomics and sensory histology techniques to analyze non-volatile and volatile compounds the second drying and pulley liquefied gas drying (PLD) samples. The results revealed that after PLD, the samples exhibited lower umami, bitterness, and astringency; whereas floral, sweet, roasted, cooked corn-like, and cooked chestnut-like aromas became stronger. Among them, the content of (−)-epigallocatechin gallate, glutamic acid, and theogallin, which were closely related to taste, decreased by 4.5 %, 12.3 %, and 10.4 %, respectively. Eight key aroma components were identified as the main contributors to the sample aroma changes: (E)- β -ionone, dimethyl sulfide, (E,E)-2,4-heptadienal, geraniol, linalool, benzeneacetaldehyde, 2-ethyl-3,5-dimethylpyrazine, and hexanal. This study provides a theoretical basis for enhancing the quality of LAGP teas. [ABSTRACT FROM AUTHOR]

Jinmudan (JM) tea, produced using Camellia sinensis, cultivar JM, is popular among consumers because of its strong floral and fruity aroma. Aroma profiles of white, green, black, and oolong teas, which were obtained from JM and Tanyangcaicha (TY, control and indigenous variety), were investigated. Sensory evaluation was used to describe the quality characteristics of the tea samples. A combination of headspace solid-phase microextraction, gas chromatography–mass spectrometry (HS-SPME/GC–MS), and chemometrics was used to extract and analyze the volatiles in the samples; 162 volatiles were identified and categorized on the basis of their chemical structures. The proportions of chemical categories differed among the teas. The same type of tea obtained from JM and TY differed significantly in concentrations and categories of volatiles. This study will improve our understanding of the profile of volatile compounds in JM teas and create a basis for investigating the biological characteristics of JM teas. [ABSTRACT FROM AUTHOR]

Objective: Adenoid cystic carcinoma (AdCC) and mucoepidermoid carcinoma (MEC) are the most frequent malignancies in the salivary glands. The purpose of this study was to explore the roles of sine oculis homeobox homolog 1 (SIX1), malic enzyme 2 (ME2), and AP‐2 complex subunit mu (AP2M1) in AdCC and MEC, as well as the potential relationship between SIX1, ME2, AP2M1, and proliferation marker cyclin D1. Material and methods: Immunohistochemistry was performed on human salivary gland tissue microarrays that contained 76 normal salivary glands (NSGs), 14 pleomorphic adenomas (PMAs), 81 AdCCs, and 52 MECs. Results: We observed that the expression levels of SIX1 and ME2 were significantly elevated in AdCC and MEC when compared with NSG and PMA. In addition, we detected that AP2M1 was overexpressed in AdCC and MEC when compared with NSG. We also found that SIX1 and AP2M1 were positively associated with cyclin D1. Conclusions: These results may prove that SIX1 and AP2M1 are involved in the proliferation of AdCC and MEC to cause tumor growth. [ABSTRACT FROM AUTHOR]

The current method for processing bud yellow tea usually results in astringent taste that is not sufficiently yellowing. Therefore, two factors in the production phase that affect tea quality were studied: number of times the tea leaves undergo the yellowing process and predrying of leaves to reduce their water content before a second yellowing process. We then evaluated the colour and nonvolatile compounds of the produced tea. Results revealed that gallated catechins and the polyphenol–amino acid ratio decreased significantly, and the colour of the tea changed from green to yellow after two yellowing processes. Predrying leaves to a lower water content resulted in the accumulation of free amino acids and soluble sugars, and an increase in the brightness and yellow colour of tea infusion. Bud yellow tea with the highest taste evaluation score and yellowness was obtained when the leave samples underwent two yellowing processes and the water content in the leaves was 40%. [ABSTRACT FROM AUTHOR]

BACKGROUND: Non‐volatile compounds play a key role in the quality and price of Keemun black tea (KBT). The non‐volatile compounds in KBT samples from different producing areas normally vary greatly. The development of rapid methods for tracing the geographical origin of KBT is useful. In this study, we develop models for the discrimination of KBT's geographical origin based on non‐volatile compounds. RESULTS: Seventy‐two KBT samples were collected from five towns in Anhui province to determine 13 KBT compounds by high‐performance liquid chromatography (HPLC). Analysis of variance showed that the content of 13 compounds in KBT indicated significant differences (P < 0.05) among five towns. Three multivariate statistical models including principal component analysis (PCA), soft independent modeling of class analogy (SIMCA), and linear discriminant analysis (LDA) were built to discriminate origin. Principal component analysis effectively extracted three principal components, namely theaflavins, galloylated catechins, and simple catechins. The high sensitivity (64.5%–99.2%) was achieved of SIMCA model. To establish the discriminant functions, six variables (gallic acid, (+)‐catechin, (−)‐epigallocatechin gallate, theaflavin‐3‐gallate, theaflavin‐3,3′‐di‐gallate, and total theaflavins) were chosen from 13 variables, and LDA was applied. This gave a satisfactory overall correct classification rate (94.4%) and cross‐validation rate (88.9%) for KBT samples. CONCLUSION: The results showed that HPLC analysis together with chemometrics is a reliable approach for tracing KBT and guaranteeing its authenticity. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Global warming has multifarious effects on crop growth and productivity. Nonetheless, the effects of moderate-high temperatures and melatonin on tea yield and quality remain unclear. In this study, we found that melatonin, a universal growth stimulatory molecule, not only promotes photosynthesis and biomass accumulation in tea plants (Camellia sinensis L.) but also improves tea quality under sub high temperature (SHT). SHT increased the dry biomass and photosynthesis by 40.8% and 28.1%, respectively, and exogenous melatonin caused a further improvement. Moreover, SHT increased the total polyphenol concentrations and decreased the free amino acid concentrations, leading to a significant increase (68.2%) in polyphenol to free amino acid ratio. However, melatonin decreased the polyphenol to free amino acid ratio by delicately improving the concentrations of polyphenols and amino acids. Consistent with the total polyphenol, melatonin increased the concentrations of (-)-catechin, (-)-gallocatechin (GC), and (-)-epigallocatechin-3-gallate (EGCG) in tea leaves. The qRT-PCR analysis revealed that melatonin increased the transcript levels of catechins biosynthesis genes, such as CsCHS, CsCH1, CsF3H, CsDFR, CsANS, CsLAR , and CsANR under SHT. Meanwhile, the theanine concentration was decreased by SHT, which was attributed to the attenuated expression of CsGS, CsGOGAT, CsGDH , and CsTS1. Nonetheless, melatonin significantly increased those transcripts and the content of theanine under SHT. Melatonin also increased the caffeine content by inducing the expression of CsTIDH, CssAMS , and CsTCS1. These results suggest that melatonin could positively alter tea growth and quality by modulating the photosynthesis and biosynthesis of polyphenols, amino acids, and caffeine in tea leaves under SHT. [ABSTRACT FROM AUTHOR]

Objectives: Dysregulation of immune cells in the tumor microenvironment is a hallmark of head and neck squamous cell carcinoma (HNSCC). Increased infiltration of pDCs has been reported in the microenvironment of HNSCC. However, the precise immunological role of pDC and the therapeutic effects of pDC depletion in HNSCC need to be further investigated.Materials and Methods: CD317 antibodies were applied for depleting pDCs in an immunocompetent transgenic HNSCC mouse model. Tumor volume was monitored. Flow cytometric analysis was conducted for studying the immune profile changes after pDC depletion. In addition, immunohistochemical staining was carried out in a human HNSCC tissue microarray for detecting the infiltration of pDCs. We also analyzed the survival implication of pDCs and its correlation with other immune related markers in human HNSCC.Results: pDC depletion in the transgenic HNSCC mouse model significantly delayed tumor growth. After pDCs were depleted, T cells were markedly revitalized, and the proportions of regulatory T cells (Tregs) and monocytic myeloid-derived suppressor cells (MDSCs) were decreased. In human HNSCC microenvironment, pDC infiltration was upregulated and its high infiltration conferred a poor prognosis. Moreover, pDC infiltration was closely correlated with the expression of Foxp-3, PD-1, TIM-3, and LAG-3.Conclusion: Our findings demonstrated that pDCs play a negative immunomodulatory role in HNSCC and may present as a target for effective immunotherapy. [ABSTRACT FROM AUTHOR]

BACKGROUND Keemun black tea (KBT) is one of the most popular tea beverages in China as a result of its unique flavor and potential health benefits. The geographical origin of KBT influences its quality and price. The present study aimed to apply a head‐space solid phase microextraction approach and gas chromatography–mass spectrometry combined with chemometric analysis to profile the volatile compounds of KBT collected from five production areas. RESULTS: Thirty‐one peaks were detected in 61 KBT samples. Hierarchical cluster analysis, principal component analysis (PCA), k‐nearest neighbor (k‐NN) and stepwise linear discriminant analysis (SLDA) were employed to visualize the volatile fractions. The results of unsupervised statistical tools were compared using a test for similarities and distinctions, which showed that different sources may be associated. A satisfying combination of average recognition (91.7%) and cross‐validation prediction abilities (84.6%) was obtained for the PCA–k‐NN. Among all of the statistical tools, SLDA provided promising results, with 100% recognition and 96.4% prediction ability. CONCLUSION: The results obtained in the present study indicate that the volatile compounds can be used as indicators to identify the geographical origin of KBT. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract Expression of the family with sequence similarity 3 member C (FAM3C) is necessary for the epithelial-mesenchymal transition (EMT). However, the expression level and clinicopathological significance of FAM3C in oral squamous cell carcinoma (OSCC) has not been thoroughly elucidated to date. We performed immunohistochemical staining on human OSCC specimens with FAM3C, co-inhibitory immune checkpoints, EMT markers, and cancer stem cells (CSCs) markers to analyze the expression levels and clinicopathological features of FAM3C in OSCC. There were 210 primary OSCC specimens, 69 oral epithelial dysplasia and 42 normal oral mucosae in our human OSCC tissue microarrays cohort. We observed that FAM3C expression was upregulated in OSCC compared with normal mucosa and epithelial dysplasia (P < 0.001). Moreover, patients with higher FAM3C expression levels had a worse prognosis than those with lower expression levels (P < 0.05). Also, FAM3C expression was positively correlated with the immune checkpoints PD-L1, VISTA, and B7-H4, the EMT marker Slug and the CSC markers SOX2 and ALDH1. In conclusion, these findings suggested that overexpression of FAM3C in human OSCC may predict a poor prognosis for OSCC patients. [ABSTRACT FROM AUTHOR]

• Increase of roasting raised odor attributes of fried, crispy rice, smoky-burnt. • The change in odor attributes is related to the content of heterocyclic compounds. • Metabolites such as amino acids and catechins contribute to forming taste of LYT. • Variations in non-volatiles and volatiles among different LYTs were elaborated. Roasting is essential for processing large-leaf yellow tea (LYT). However, the effect of the roasting on the metabolic and sensory profiles of LYT remains unknown. Herein, the metabolomics and sensory quality of LYT at five roasting degrees were evaluated by liquid/gas chromatography mass spectrometry and quantitative descriptive analysis. A higher degree of roasting resulted in a significantly stronger crispy rice, fried rice, and smoky-burnt aroma (p < 0.05), which is closely associated with heterocyclic compound accumulation (concentrations: 6.47 ± 0.27 – 1065.00 ± 5.58 µg/g). Amino acids, catechins, flavonoid glycosides and N -ethyl-2-pyrrolidone-substituted flavan-3-ol varied with roasting degree. The enhancement of crispy-rice and burnt flavor coupled with the reduction of bitterness and astringency. Correlations analysis revealed the essential compounds responsible for roasting degree, including 2,3-diethyl-5-methylpyrazine, hexanal, isoleucine, N -ethyl-2-pyrrolidone-substituted flavan-3-ol (EPSF), and others. These findings provide a theoretical basis for improving the specific flavors of LYT. [ABSTRACT FROM AUTHOR]

Withering leads to the formation of high-quality flavor in black tea. This study investigated the effects of natural withering (NW), sun withering (SW), and warm-air withering (WW) on the flavor compounds of withered leaves by using target metabolomic and transcriptomic approaches. The results revealed that 4282, 4770, and 1246 differentially expressed genes (DEGs) were identified in the NW, SW, and WW samples, respectively. The expression of DEGs involved in volatile compound and flavonoid biosynthesis was the highest in the WW sample, which may be the main reason for the significant increase in volatile compound and catechin contents in the WW sample. The withering methods mainly affected the contents of non-proteinaceous amino acids, such as theanine. Withering methods significantly affect the expression levels of DEGs involved in metabolic pathways; this provides a systematic explanation for flavor formation in black tea. • Different withering methods affect the flavor formation of black tea. • The essence of the withering method is the type of stress. • Warm-air withering increased the content of flavor compounds. • Comprehensive explanation of the mechanism of action of withering method of black tea. [ABSTRACT FROM AUTHOR]

[Display omitted] • Over-fired drying methods affect the aroma quality of Lu'an Guapian (LAGP). • (E)- β -ionone is the most crucial aroma active compound in LAGP. • Pulley charcoal drying significantly enhanced the floral and fruity aroma of LAGP. Over-fired drying, a crucial process in the production of Lu'an Guapian (LAGP) tea, greatly enriches the tea's aroma. In this study, the aroma compounds of LAGP tea processed through pulley charcoal drying (PCD), roller drying (RD), roller-conveyor drying (RCD), and hot air drying (HD) were analyzed using gas chromatography–mass spectrometry. A subsequent analysis of aroma extraction dilution analysis and odor activity values revealed that (E)- β -ionone, dimethyl sulfide, (E,E)-2,4-heptadienal, geraniol, linalool, benzeneacetaldehyde, coumarin, 2-ethyl-3,5-dimethyl-pyrazine, indole, hexanal, (Z)-jasmone, and (Z)-3-hexen-1-ol were the key contributors to the samples' aroma variation. Moreover, a quantitative descriptive analysis and aroma recombination and omission experiments analysis revealed that (E)- β -ionone is the most critical contributor to the formation of floral aroma in tea processed using PCD, whereas (E,E)-2,4-heptadienal is responsible for the more pronounced fresh aroma in tea processed using HD. In addition, 2-ethyl-3,5-dimethyl-pyrazine contributes to the formation of a roasted aroma in tea processed using RD and RCD. The study results provide a theoretical basis for choosing the processing method, especially for drying, to obtain high-quality LAGP tea. [ABSTRACT FROM AUTHOR]

The immune system plays a critical role in the establishment, development, and progression of head and neck squamous cell carcinoma (HNSCC). As treatment with single-immune checkpoint agent results in a lower response rate in patients, it is important to investigate new strategies to maintain favorable anti-tumor immune response. Herein, the combination immunotherapeutic value of CTLA4 blockade and SFKs inhibition was assessed in transgenic HNSCC mouse model. Our present work showed that tumor growth was not entirely controlled when HNSCC model mice were administered anti-CTLA4 chemotherapeutic treatment. Moreover, it was observed that Src family kinases (SFKs) were hyper-activated and lack of anti-tumor immune responses following anti-CTLA4 chemotherapeutic treatment. We hypothesized that activation of SFKs is a mechanism of anti-CTLA4 immunotherapy resistance. We, therefore, carried out combined drug therapy using anti-CTLA4 mAbs and an SFKs’ inhibitor, dasatinib. As expected, dasatinib and anti-CTLA4 synergistically inhibited tumor growth in Tgfbr1/Pten 2cKO mice. Furthermore, dasatinib and anti-CTLA4 combined to reduce the number of myeloid-derived suppressor cells and Tregs, increasing the CD8+ T cell-to-Tregs ratio. We also found that combining dasatinib with anti-CTLA4 therapy significantly attenuated the expression of p-STAT3Y705 and Ki67 in tumoral environment. These results suggest that combination therapy with SFKs inhibitors may be a useful therapeutic approach to increase the efficacy of anti-CTLA4 immunotherapy in HNSCC. [ABSTRACT FROM AUTHOR]

Abstract: A major challenge for traditional cancer therapy, including surgical resection, chemoradiotherapy, and immunotherapy, is how to induce tumor cell death and leverage the host immune system at the same time. Here, a myeloid‐derived suppressor cell (MDSC) membrane‐coated iron oxide magnetic nanoparticle (MNP@MDSC) to overcome this conundrum for cancer therapy is developed. In this study, MNP@MDSC demonstrates its superior performance in immune evasion, active tumor‐targeting, magnetic resonance imaging, and photothermal therapy (PTT)‐induced tumor killing. Compared with red blood cell membrane‐coated nanoparticles (MNPs@RBC) or naked MNPs, MNP@MDSCs are much more effective in active tumor‐targeting, a beneficial property afforded by coating MNP with membranes from naturally occurring MDSC, thus converting the MNP into “smart” agents that like to accumulate in tumors as the source MDSCs. Once targeted to the tumor microenvironment, MNPs@MDSC can act as a PTT agents for enhanced antitumor response by inducing immunogenic cell death, reprogramming the tumor infiltrating macrophages, and reducing the tumor's metabolic activity. These benefits, in combination with the excellent biocompatibility and pharmacological kinetics characteristics, make MNP@MDSC a promising, multimodal agent for cancer theranostics. [ABSTRACT FROM AUTHOR]

Objectives: The aims of this study were to investigate the relationship between Golgi phosphoprotein 2 (GOLPH2) and oral squamous cell carcinoma (OSCC) and explore the clinical significance of GOLPH2 in OSCC.Methods: Tissue microarrays from human OSCC samples were stained for GOLPH2 expression and clinicopathologic features. Kaplan-Meier analysis was used to compare the survival of patients with high GOLPH2 expression and patients with low GOLPH2 expression.Results: We found GOLPH2 is highly expressed in OSCC tissue, and the GOLPH2 expression in metastatic lymph nodes is higher than in tumor tissue. Our data indicate that patients with higher GOLPH2 expression have poor overall survival compared with those with lower GOLPH2 expression. This study demonstrated that GOLPH2 was associated with CD44, SOX2, Slug, B7-H3, B7-H4, TIM3, and VISTA.Conclusions: These findings suggest GOLPH2 is a potential marker for estimating the patient's prognosis and may be a target for molecular-targeted therapy against OSCC. [ABSTRACT FROM AUTHOR]

The NLRP3 inflammasome is a critical innate immune pathway responsible for producing active interleukin (IL)-1β, which is associated with tumor development and immunity. However, the mechanisms regulating the inflammatory microenvironment, tumorigenesis and tumor immunity are unclear. Herein, we show that the NLRP3 inflammasome was over-expressed in human HNSCC tissues and that the IL-1β concentration was increased in the peripheral blood of HNSCC patients. Additionally, elevated NLRP3 inflammasome levels were detected in tumor tissues of Tgfbr1/Pten 2cKO HNSCC mice, and elevated IL-1β levels were detected in the peripheral blood serum, spleen, draining lymph nodes and tumor tissues. Blocking NLRP3 inflammasome activation using MCC950 remarkably reduced IL-1β production in an HNSCC mouse model and reduced the numbers of myeloid-derived suppressor cells (MDSCs), regulatory T cells (Tregs) and tumor-associated macrophages (TAMs). Moreover, inhibiting NLRP3 inflammasome activation increased the numbers of CD4+ and CD8+ T cells in HNSCC mice. Notably, the numbers of exhausted PD-1+ and Tim3+ T cells were significantly reduced. A human HNSCC tissue microarray showed that NLRP3 inflammasome expression was correlated with the expression of CD8 and CD4, the Treg marker Foxp3, the MDSC markers CD11b and CD33, and the TAM markers CD68 and CD163, PD-1 and Tim3. Overall, our results demonstrate that the NLRP3 inflammasome/IL-1β pathway promotes tumorigenesis in HNSCC and inactivation of this pathway delays tumor growth, accompanied by decreased immunosuppressive cell accumulation and an increased number of effector T cells. Thus, inhibition of the tumor microenvironment through the NLRP3 inflammasome/IL-1β pathway may provide a novel approach for HNSCC therapy. [ABSTRACT FROM AUTHOR]

Phenolic compounds, such as flavanols, flavones, flavonols, anthocyanins, and phenolic acids, contribute to a plant’s adaptation to their ecological niches. These compounds serve as attractants of organisms beneficial to the plants or as defence against other biotic and abiotic factors during the plant evolution. In this paper, the accumulation patterns of phenolic compounds in the eight evergreen woody core eudicots indicated divergent and convergent evolution of plant secondary metabolites. The results indicated some phenolic acids and flavonols were distributed in all the studied eight evergreen woody plants, within Oleaceae, Theaceae, Caprifoliaceae and Celastraceae, with different accumulation profiles. The flavan-3-ols, monomeric catechins, and polymeric proanthocyanidins, were undetectable in Osmanthus fragrans , Ligustrum lucidum, and Fraxinus mariesii of Oleaceae, but highly accumulated in the other five species, especially in Camellia sinensis of Theaceae. It indicated Camellia sinensis evolved great ecological adaptive capacity to prevent herbivores, insect pests, and microbial pathogens. [ABSTRACT FROM AUTHOR]

Nanoparticles possess the potential to revolutionize cancer diagnosis and therapy. The ideal theranostic nanoplatform should own long system circulation and active cancer targeting. Additionally, it should be nontoxic and invisible to the immune system. Here, the authors fabricate an all-in-one nanoplatform possessed with these properties for personalized cancer theranostics. Platelet-derived vesicles (PLT-vesicles) along with their membrane proteins are collected from mice blood and then coated onto Fe3O4 magnetic nanoparticles (MNs). The resulting core-shell PLT-MNs, which inherit the long circulation and cancer targeting capabilities from the PLT membrane shell and the magnetic and optical absorption properties from the MN core, are finally injected back into the donor mice for enhanced tumor magnetic resonance imaging (MRI) and photothermal therapy (PTT). Meanwhile, it is found that the PTT treatment impels PLT-MNs targeting to the PTT sites (i.e., tumor sites), and exactly, in turn, the enhanced targeting of PLT-MNs to tumor sites can improve the PTT effects. In addition, since the PLT membrane coating is obtained from the mice and finally injected into the same mice, PLT-MNs exhibit stellar immune compatibility. The work presented here provides a new angle on the design of biomimetic nanoparticles for personalized diagnosis and therapy of various diseases. [ABSTRACT FROM AUTHOR]

[Display omitted] • IDA was composed of phenylboronic acids, pyrocatechol violet, and alizarin red S. • TP, TC, and EGCG were key quality indicators of black tea fermentation. • SVM model achieved accurate qualitative evaluation of fermentation degrees. • SCARS-SVR model achieved accurate prediction of TP, TC, and EGCG. • In-situ quality monitoring of black tea fermentation was realized. Herein, a new indicator-displacement array (IDA) sensor was developed for the quality evaluation of black tea fermentation. On the principle of the reversible covalent binding of phenylboronic acid and catechol, phenylboronic acids were selected as acceptors for targeted binding to polyphenols. Pyrocatechol violet and alizarin red S were used as indicators of the reaction. The IDA sensors have sensitive differential responses to fermented tea samples, achieving an assessment of the fermentation degree with accuracies of 80.39–88.00% by support vector machine (SVM). In addition, the key polyphenol components of the fermentation process were accurately predicted by the IDA and SVM regression with ratio of prediction to deviation values of 1.55–1.72, 2.03–2.21, and 2.03–2.08 for total polyphenols, total catechins, and epigallocatechin-3-O-gallate, respectively. In conclusion, the developed IDA sensor is capable of the in-situ quality monitoring of black tea fermentation, with the advantages being cost-effectiveness, sensitivity, and rapidity. [ABSTRACT FROM AUTHOR]

Fixation is the key step of green tea processing, usually with moisture content as the quality evaluation indicator. Near-infrared (NIR) spectroscopy has been widely used in tea moisture detection but is mostly realized in the laboratory, and online detection in tea processing is limited. This study explores the possibility of calibration transfer from lab-based hyperspectral imaging (HSI) to online NIR to enable the rapid detection of moisture during the fixation step. Raw spectral data were preprocessed, and direct standardization (DS) and partial least-squares regression (PLS) were used for data calibration and modeling, respectively. The results showed that there were observable differences in the spectral data of the same sample obtained by different instruments. The prediction model based on HSI failed to be transferred directly to the online NIR and has a large root mean square error of prediction (RMSEP) value. Spectral preprocessing with a standard normal variate (SNV) transformation reduced the differences between the instruments. When 40 standard samples were used, the established transferable SNV-DS-PLS model based on HSI achieved the optimal prediction performance with a correlation coefficient of prediction (Rp), RMSEP, and ratio of prediction to deviation (RPD) of 0.94, 2.24%, and 2.76, respectively, for the 50 validated tea samples obtained from online NIR. In conclusion, the model transfer across instruments was successfully realized, and tea moisture detection was brought from the laboratory to the tea factory, which provided a reference model for online moisture detection for other agricultural products. • Calibration transfer from HSI to online NIR for tea processing was optimized. • Online detection of moisture during green tea fixation was achieved. • Model built based on HSI fails to be transferred directly to the online NIR. • Transferable SNV-DS-PLS model gave the optimal performance for moisture. [ABSTRACT FROM AUTHOR]

• Data of fermented tea samples was collected at-line by an industrial camera. • Catechins, theaflavins and chlorophylls were chosen as key quality indicators. • Correlations between color variables and quality indicators were established. • Spatial and temporal distribution of indicators was mapped and visualized. Fermentation is a key black tea processing step and makes an important contribution to quality formation. Current approaches to fermentation monitoring are costly or laboratory-based. Here, we first evaluated the potential of at-line computer vision for detecting fermentation quality in a tea factory. A self-built industrial camera was used to collect tea samples at various fermentation durations. The correlations of color variables that were extracted from the images with key quality indicators in the tea samples were verified. Subsequently, partial least-squares regression models based on the color variables showed high prediction accuracy with residual prediction deviation values of 4.13, 3.53, and 3.39 for catechins, theaflavins and chlorophylls, respectively. Finally, the spatial and temporal distributions of indicators during fermentation were mapped to visualize the fermentation quality. This study realized low-cost, at-line and real-time detection for black tea fermentation, which provides technical support for the industrial and intelligent production of black tea. [ABSTRACT FROM AUTHOR]

Cumulative evidence suggests that constitutively activated signal transducer and activator of transcription (STAT3) may contribute to sustaining immunosuppressive status, and that inhibiting STAT3 signaling represents a potential strategy to improve antitumor immunity. In the present study, we observed that high levels phosphorylated of STAT3 are significantly associated with the markers for both myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs) in human head and neck squamous cell carcinoma (HNSCC). Additionally, we showed that targeting STAT3 signaling with a tolerable selective inhibitor S3I-201 significantly decreased immature myeloid cells such as MDSCs, TAMs and iDCs in genetically defined mice HNSCC model. These findings highlight that targeting STAT3 signaling may be effective to enhance antitumor immunity via myeloid suppressor cells in HNSCC. [ABSTRACT FROM AUTHOR]

• Different withering methods affect the aroma quality of Keemun black tea. • Sun withering significantly enhanced the floral and fruity of Keemun black tea. • Methional, geraniol and linalool are the most active-aroma in Keemum black tea. • A more comprehensive profile of Keemun aroma was obtained. Withering is a key process that affects the aroma of Keemun black tea (KBT). In this study, the aroma composition of KBT through natural withering, sun withering, and warm-air withering was analysed using gas chromatography–mass spectrometry. The results revealed significant differences in the three samples. Gas chromatography–olfactometry and aroma extract dilution analysis were performed with screening through a relative odour activity value (rOAV) > 1. In total, 11 aroma-active compounds (geraniol, (Z)-4-heptenal, 1-octen-3-ol, (E)- β -ionone, 3-methylbutanal, linalool, β -damascenone, (E , E)-2,4-decadienal, methional, (E , E)-2,4-nonadienal, and (E)-2-nonenal) were found to be responsible for the differences in aroma caused by different withering methods. Linalool (rOAV, 161) and geraniol (rOAV, 785) were responsible for the higher flowery and fruity aromas when sun withering was applied, whereas methional (rOAV, 124) contributed to the intense roasty aroma when warm-air withering was employed. Moreover, our results were verified by quantitative descriptive analysis and addition experiments. [ABSTRACT FROM AUTHOR]

• DNA methylation is the most common epigenetic modification that plays a crucial role in cancer tumorigenesis, progression, and therapeutic resistance. • The high expression of DNMT1 in OSCC may predict an immunosuppressive tumor microenvironment (TME) and a worse prognosis in OSCC. • DNMT1 inhibitor treatment may improve anti-tumor immunity by suppressing the tumor immune microenvironment. Epigenetic alterations, including DNA methylation, play crucial roles in the tumor. Epigenetic drugs like DNA methyltransferase-1 (DNMT1) inhibitors have been exhibited positive effects in cancer treatment. However, the role of DNMT1 in oral squamous cell carcinoma (OSCC) is less clearly described. What is more, the effects on the immune microenvironment of DNMT1 have not become appreciated. In this research, we determine the expression levels of DNMT1 and the association of prognosis by analyzing human OSCC tissue microarrays. Two different types of immunocompetent mouse OSCC models were established to explore the effects of DNMT1 inhibitor on the tumor microenvironment(TME). We identified DNMT1 was highly expressed both in human and mouse OSCC tissues. The expression levels of DNMT1 was also correlated with the immunosuppressive molecules and tumor-promoter such as VISTA, PD-L1, B7-H4, and PAK2, indicating a worse prognosis. Of particular concern is that DNMT1 inhibition improved TME and delayed tumor growth by decreasing myeloid-derived suppressor cells (MDSCs) and increasing tumor-infiltrating T cells. Our data suggests that DNMT1 play a key role in OSCC and has a possible immunotherapeutic marker treatment. [ABSTRACT FROM AUTHOR]

[Display omitted] • We evaluated the genes involved in catechins biosynthesis in C. sinensis. • In total, 47,717 unigenes were obtained from two cultivars of C. sinensis. • KEGG database analysis identified 13 key enzymes involved in catechins biosynthesis. • The structures of LAR and ANR were spatially modeled. • Our findings established a basis for further studies of catechins biosynthesis. Camellia sinensis (L.) O. Kuntze is used to produce tea, a beverage consumed worldwide. Catechins are major medically active components of C. sinensis and can be used clinically to treat hyperglycaemia, hypertension, and cancer. In this study, we aimed to identify the genes involved in catechins biosynthesis. To this end, we analysed transcriptome data from two different cultivars of C. sinensis using DNBSEQ technology. In total, 47,717 unigenes were obtained from two cultivars of C. sinensis , of which 9429 were predicted as new unigenes. In our analyses of the Kyoto Encyclopedia of Genes and Genomes database, 212 unigenes encoding 13 key enzymes involved in catechins biosynthesis were identified; the structures of leucoanthocyanidin reductase and anthocyanidin reductase were spatially modelled. Some of these key enzymes were verified by real-time quantitative polymerase chain reaction, and multiple genes encoding plant resistance proteins or transcription factors were identified and analysed. Furthermore, two microRNAs involved in the regulation of catechins biosynthesis were explored. Differentially expressed genes involved in the flavonoid biosynthesis pathway were identified from pairwise comparisons of genes from different cultivars of tea plants. Overall, our findings expanded the number of publicly available transcript datasets for this valuable plant species and identified candidate genes related to the biosynthesis of C. sinensis catechins, thereby establishing a foundation for further in-depth studies of catechins biosynthesis in varieties or cultivars of C. sinensis. [ABSTRACT FROM AUTHOR]

• JMD black tea aroma profiles and its characteristic aromas were found. • JMD black tea has a strong fruity flavor. • Jasmine lactone is the most important fruity flavor substance. • Tea cultivar affect the aroma formation. Jinmudan (JMD) black tea is processed from the JMD cultivar (Camellia sinensis (L.) O. Kuntze cv. Jinmudan). Dried JMD black tea (JMD-D) received a score of 7.43 in the quantitative descriptive analysis, indicating that it had the strongest fruity flavor. Many volatile compounds (129) were identified by solvent assisted flavor evaporation and gas chromatograph mass spectrometry. In total, 26 aroma active compounds, including (Z)-methyl epi-jasmonate (flavour dilution factor (FD): 128–1024), benzeneacetaldehyde (FD: 512–1024), linalool (FD: 256–1024), geraniol (FD: 256–512), jasmine lactone (FD: 64–1024), were identified in JMD-D and controls through an aroma extract dilution analysis and gas chromatography-mass spectrometry olfactometry. On the basis of the FD values, odor activity values, and relative quantification results, the volatiles of fruit flavor derived from lipids, such as jasmine lactone, were prominent in JMD-D compared to the controls. During the processing of the four types of black tea, the volatile fraction concentration increased mainly from the fresh leaf to the fermentation stage and decreased in the drying stage. [ABSTRACT FROM AUTHOR]

• An ecologically insensitive variegated tea plant was studied in this research. • The component, tissue structure, gene and protein expressions were analyzed. • Reduced catechins and increased amino acids were found in variegated tea leaves. • Related genes and proteins were down-regulated in flavonoid biosynthetic pathway. • The high expression of CLH1, showed an active chlorophyll degradation in variegated leaves. Globally, tea plant (Camellia sinensis (L.) O. Kuntze) is a crucial commercial crop. In this study, a variegated tea plant identified in Huangshan, China. The variegated and residual normal leaves were used as the research material. The variegated leaves of the plant were ecologically insensitive. The components, tissue structure, gene expression patterns, and protein expression profiles of the variegated tea plant were analyzed. Ultrastructural analysis of the variegated leaves revealed that the chloroplasts of the leaves were abnormally developed, with no obvious thylakoids. Compared with normal leaves, the albino leaves had lower total catechins content and higher total free amino acid content. Through RNA sequencing and proteomic analysis, chlorophyll biosynthesis and degradation, photosynthesis, chloroplast development, and flavonoid biosynthesis of the variegated leaves were affected. The less chlorophyll content in the variegated tea leaves could probably attributed to the destruction of chloroplast structure. Besides, the relatively high expression and enzyme activity of CLH, showed an active chlorophyll degradation; the lower expression of Lhcb1, Lhcb3 ; and POR in chlorophyll synthesis, might be related to the low chlorophyll in variegated leaves. In terms of flavonoid biosynthesis, expression of the related genes was down-regulated and the target protein content of ANR, CHI) was also reduced. These results may provide insight into the catechins mechanism of such variegated phenotypes. [ABSTRACT FROM AUTHOR]

C-repeat/dehydration-responsive element binding factors (CBFs) can induce the expression of a suite of cold-responsive genes to increase plant cold tolerance, and inducer of CBF expression 1 (ICE1) is a major activator for CBF. In the present study, we isolated the full-length cDNAs of ICE1 and CBF from Camellia sinensis, designated as CsICE1 and CsCBF1, respectively. The deduced protein CsICE1 contains a highly conserved basic helix-loop-helix (bHLH) domain and C-terminal region of ICE1-like proteins. CsCBF1 contains all conserved domains of CBFs in other plant species and can specifically bind to the C-repeat/dehydration-responsive element (CRT/DRE) as confirmed by electrophoretic mobility shift assay. The transcription of CsICE1 had no apparent alteration after chilling treatment (4°C). CsCBF1 expression was not detected in normal temperature (20°C) but was induced immediately and significantly by low temperature (4°C). Our results suggest that ICE1-CBF cold-response pathway is conserved in tea plants. CsICE1 and CsCBF1, two components of this pathway, play roles in cold responses in tea plants. [ABSTRACT FROM AUTHOR]

• Low-cost, portable instruments were used to evaluate the tea processing quality. • A satisfactory classification rate (89.19%) was achieved using data fusion. • Quantitative model showed high prediction performance for the theaflavin content. • Our results provide templates for at-line monitoring of tea processing quality. Intelligent identification of black tea fermentation quality is becoming a bottleneck to industrial automation. This study presents at-line rapid detection of black tea fermentation quality at industrial scale based on low-cost micro-near-infrared spectroscopy (NIRS) and laboratory-made computer vision system (CVS). High-performance liquid chromatography and a spectrophotometer were used for determining the content of catechins and theaflavins, and the color of tea samples, respectively. Hierarchical cluster analysis combined with sensory evaluation was used to group samples through different fermentation degrees. A principal component analysis–support vector machine (SVM) model was developed to discriminate the black tea fermentation degree using color, spectral, and data fusion information; high accuracy (calibration = 95.89%, prediction = 89.19%) was achieved using mid-level data fusion. In addition, SVM model for theaflavins content prediction was established. The results indicated that the micro-NIRS combined with CVS proved a portable and low-cost tool for evaluating the black tea fermentation quality. [ABSTRACT FROM AUTHOR]

Withering, or "spreading" in Chinese word, is a key process in Chinese green tea production. However, the underlying mechanisms of this treatment remains unclear. To investigate the effect of withering treatment on the quality of postharvest tea leaves, UHPLC-Orbitrap-MS metabolomics and tandem mass tag (TMT)-based proteomics analysis were performed. A total of 69 metabolites were identified, and up to 136 different expressed proteins were obtained through pair-wise comparisons. Comprehensive metabolomics and proteomics analyses revealed a monotonic metabolic shift ascribed to the tea leaves' response to dehydration stress caused by harvesting. Multiple amino acids were increased in level due to protein degradation; the biosynthesis of catechins and flavonoids were inhibited; the level of glycosidically bound volatiles were decreased attributed to the metabolism enhancement. These metabolic changes provide a material basis for later processes in green tea production and help with the quality formation of green tea. • Metabolomics and proteomics were used for tea withering study. • A comprehensive view was acquired by combining metabolomics and proteomics results. • Amino acids and glycosidically bound volatiles were mostly affected during withering. • Withered leaves present a monotonic metabolic shift ascribed to multiple stresses. [ABSTRACT FROM AUTHOR]