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Three new α-pyrone derivatives cytospotones A-C (1-3) and a new cyclohexenone derivative cytospotone D (4) together with four known α-pyrones were isolated from the endophytic fungus Cytospora sp. A879 of Pogostemon cablin (Blanco) Benth. The structures of 1-4 were elucidated primarily by spectroscopic methods (1D, 2D NMR and HRESIMS), ECD spectra analyses, and ECD calculations. Furthermore, the four new compounds (1-4) were evaluated for their anti-inflammatory and α-glucosidase inhibitory activities. The results showed that compound 1 had moderate inhibitory effect on LPS-induced NO production in RAW 264.7 macrophages., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Post-translational modifications (PTMs), such as phosphorylation and ubiquitination, play key roles in signal transduction and protein homeostasis. The crosstalk of PTMs greatly expands the components of proteome and protein functions. Multi-level proteome analysis, which involves proteome investigations of total lysate and PTMs in this context, provides a comprehensive approach to explore the PTM crosstalk of a biological system under diverse disturbances. However, multi-level proteome practice remains technically challenging. Here we intended to build a strategy for multi-level proteome analysis, in which we focus on the serial profiling the total proteome, ubiquitinome and phosphoproteome from the microscale of starting material. We started by evaluating five common lysis buffers and found that the sodium deoxycholate buffer provided the best overall performance. We then developed an approach for serial enrichment and profiling of the multi-level proteome. To expand the depth of identification, we customized the variable windows to perform data-independent acquisition (DIA) sequencing for each proteome. In total, we identified 6465 proteins, ∼20,000 GlyGly sites (class 1), and ∼ 19,000 phosphosites (class 1) sequentially using 1 mg of HeLa digest by three DIA measurements. We applied this strategy to analyze MG132-treated HeLa cells and observed the crosstalk between ubiquitination and phosphorylation. Our method can be referenced for other multi-level proteome studies with microscale samples. SIGNIFICANCE: Lysis buffer containing sodium deoxycholate provided the best overall performance in multi-level proteome analysis. One step of ubiquitination enrichment before phosphorylation enrichment does not reduce the reproducibility of phosphoproteome. Customized isolation windows were established for DIA analysis on each level of proteome. Combined the serial enrichment approach and the customized single-shot DIA method enabled the multi-level proteome of microscale protein samples., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Climate change has substantially affected plant phenology and growth on the Qinghai-Tibetan Plateau (QTP), while it remains unclear how plant phenology and growth impact the plant biomass under climate change. We used long-term data (from 1997 to 2017) for four plants, Stipa purpurea, Artemisia scoparia, Kobresia humilis and Astragalus laxmannii in the alpine meadow to examine the relationships among multiple climate factors, vegetative growth, reproductive growth, intrinsic growth rate and biomass. The order of returning to green determines the growth strategies of different plants, the earliest plants to green (p < 0.05) (e.g., Stipa purpurea and Artemisia scoparia) would choose the strategy of vegetative growth (p < 0.05); the earlier plants (p < 0.05) (e.g., Kobresia humilis) would be regulated by both vegetative growth and reproductive growth (p < 0.05); while the latest plant to green (p < 0.05) such as Astragalus laxmannii, would choose intrinsic growth rate rather than growing season (P < 0.05). Temperature was the most important drivers for key phenological phases and growth patterns of four species, different factors play a role in different stages of the growth period, i.e., in the early and late stage is the soil temperature, while in the middle stage is the average temperature or the maximum temperature, and all the optimum thresholds were >30 day. These findings provide the in-situ evidences of long-term changes in phenology and its associated growth on the biomass of alpine plants on the QTP in the era of climate change., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Increased nitrogen (N) deposition can affect ecosystem processes and thus influence plant eco-physiological processes in grasslands. However, how N deposition affects eco-physiological processes of leguminous and non-leguminous forbs in alpine grasslands is understudied. A long-term field experiment using a range of simulated N deposition rates (0, 8, 24, 40, 56, and 72 kg N ha -1  year -1 ) was established to examine the effects of N deposition on various eco-physiological parameters in leguminous and non-leguminous forbs in an alpine meadow of the Qinghai-Tibetan Plateau. We found that the responses of leguminous and non-leguminous forbs to simulated N deposition varied. Net photosynthetic rate of leguminous and non-leguminous forbs exhibited different response patterns, but chronic increases in simulated N deposition rates may lead to negative effects in both functional groups. Neither functional group responded differently in aboveground biomass under the highest N addition level (72 kg N ha -1  year -1 ) compared to the control. Differences in aboveground biomass of leguminous forbs were observed at intermediate N levels. Short-term simulated N deposition significantly promoted N uptake of both functional groups. In leguminous forbs, simulated N deposition affected net photosynthetic rates (P N ) and aboveground biomass (AGB) mainly via stomatal conductance (gs), water use efficiency (WUE), and plant N uptake. In non-leguminous forbs, simulated N deposition affected P N and AGB mainly through WUE and plant N uptake. Our findings suggest that leguminous and non-leguminous forbs have differential response mechanisms to N deposition, and compared with non-leguminous forbs, leguminous forbs are more sensitive to continuing increased N deposition. The obvious decline trend in photosynthetic capacity in leguminous forbs is likely to exacerbate the already divergent ecological processes between leguminous and non-leguminous forbs. More importantly, these changes are likely to alter the future composition, function, and stability of alpine meadow ecosystems., Competing Interests: Declaration of competing interest No conflict of interest exists in the submission of this manuscript, and the manuscript is approved by all authors for publication. I would like to declare on behalf of my co-authors that the work described was original research that has not been published previously, and not under consideration for publication elsewhere, in whole or in part. All the authors listed have approved the manuscript that is enclosed., (Copyright © 2020. Published by Elsevier B.V.)

Qinghai-Tibetan Plateau (QTP), >4000 m known as the "third pole of the earth" and is highly sensitive to nitrogen (N) deposition, understanding the effects of N deposition on multifaceted plant diversity (taxonomy diversity, functional diversity and phylogenetic diversity) in the alpine grasslands of Qinghai-Tibet Plateau are vital for the conservation of alpine plant diversity and the sustainability of alpine grasslands ecosystem services. We added N of different gradients to test the effects of soil acidification, soil eutrophication, and phosphorus limitation independently, and interactively on the multifaceted plant richness and evenness in both an alpine meadow and an alpine steppe of the QTP. We found that all the p-value of taxonomy diversity, functional diversity and phylogenetic diversity were >0.05 and values of R 2 of fixed factors by nitrogen addition gradients was low (<0.10). In contrast to the alpine steppe, diversity of alpine meadow is more sensitive to soil factors than alpine steppe. Soil acidification caused by nitrogen deposition changed taxonomic evenness (p < 0.05), while eutrophication associated with nitrogen deposition altered taxonomic richness and phylogenetic evenness (p < 0.05) in the alpine meadow and functional richness (p < 0.05) in the alpine steppe. These findings suggest that the effects of N deposition on the multifaceted plant diversity (taxonomic, functional and phylogenetic diversity) varied with N deposition gradients and ecosystem types. Rational adaptation and mitigation techniques should be considered for different types of alpine grasslands on the QTP according to their different responses to the nitrogen deposition gradients in the future., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Peroxisome proliferator-activated receptor alpha (PPARα) controls lipid homeostasis through regulation of lipid transport and catabolism. PPARα activators are clinically used for hyperlipidemia treatment. The role of PPARα in bile acid (BA) homeostasis is beginning to emerge. Herein, Ppara-null and hepatocyte-specific Ppara-null (Ppara ∆Hep ) as well as the respective wild-type mice were treated with the potent PPARα agonist Wy-14,643 (Wy) and global metabolomics performed to clarify the role of hepatocyte PPARα in the regulation of BA homeostasis. Levels of all serum BAs were markedly elevated in Wy-treated wild-type mice but not in Ppara-null and Ppara ∆Hep mice. Gene expression analysis showed that PPARα activation (1) down-regulated the expression of sodium-taurocholate acid transporting polypeptide and organic ion transporting polypeptide 1 and 4, responsible for the uptake of BAs into the liver; (2) decreased the expression of bile salt export pump transporting BA from hepatocytes into the bile canaliculus; (3) upregulated the expression of multidrug resistance-associated protein 3 and 4 transporting BA from hepatocytes into the portal vein. Moreover, there was a notable increase in the compositions of serum, hepatic and biliary cholic acid and taurocholic acid following Wy treatment, which correlated with the upregulated expression of the Cyp8b1 gene encoding sterol 12α-hydroxylase. The effects of Wy were identical between the Ppara ∆Hep and Ppara-null mice. Hepatocyte PPARα controlled BA synthesis and transport not only via direct transcriptional regulation but also via crosstalk with hepatic farnesoid X receptor signaling. These findings underscore a key role for hepatocyte PPARα in the control of BA homeostasis., (Published by Elsevier B.V.)

Lithocarols A-F (1-6) possessing novel highly-oxygenated isobenzofuran core, together with a related known compound isoprenylisobenzofuran A (7) were isolated from the marine-derived fungus Phomopsis lithocarpus FS508. Among them, lithocarols A-E (1-5) represent the first examples of poly-ketal derivatives in tenellone family. The structures for all these compounds were fully elucidated by spectroscopic analysis, X-ray diffraction, and electronic circular dichroism calculations. Their cytotoxic assay disclosed that compounds 1-4 displayed moderate growth inhibitory effect against four human tumor cell lines with the IC 50 values ranging from 10.5 to 38.7 μM., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Temperate desert is one of the globally important biomes with unique and valuable biodiversity, which might be threatened by environmental stresses and human disturbance associated with rapid development. However, few studies have documented the spatial distribution of the multifaceted plant diversity of the temperate desert and their relationships with external impacting factors. We sampled multifaceted plant species diversity including taxonomic diversity, functional diversity and phylogenetic diversity in the Alashan Desert along Beijing-Xinjiang Expressway (G6) in Northern China to identify the key factors and process which regulate the multifaceted plant diversity of the temperate desert. We found that the dynamics of species richness, functional richness, and phylogenetic richness along the elevational gradient corresponded to the unimodal model. Species phylogenetic development shifted from aggregation to divergence, while species functional traits were the opposite along the elevational gradient. The sites at an elevation around 1200-1400 m were the key habitats for the occurrence of high plant diversity including species richness, functional richness and phylogenetic richness. There were no significant differences (p > 0.05) in plant diversity at different distances from the road (500 m, 1000 m and 1500 m) and human disturbances (the distance from the nearest human settlements). Temperature, temperature variability, precipitation, precipitation variability, soil physical and chemistry properties showed no significant effects on plant diversity. It was concluded that evolutionary history and functional traits, not environmental or anthropogenic factors were the key determinants of the pattern of multifaceted plant diversity., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Sesquiterpene synthases are key enzymes for biosynthesis of sesquiterpene compounds and are important for agarwood formation in Aquilaria sinensis.The As-sesTPS gene encoding a novel sesquiterpene synthase was expressed in Escherichia coli strain BL21 (DE3) as an inclusion body and purified by Ni affinity chromatography. The molecular weight of the protein was lower than the theoretical value. Amino acid sequencing results indicated that the 27.2kDa-recombinant protein was a truncated sesquiterpene synthase from chemically induced A. sinensis. After refolding, the truncated As-SesTPS protein catalyzed the conversion of farnesyl pyrophosphate (FPP) to nerolidol which is a characteristic component of agarwood. The optimal reaction pH for the As-SesTPS protein was 8.0, and the optimal temperature was 30°C. The values of Km and Vmax of As-SesTPS protein towards FPP were 0.0548mM, 42.83μmol/mg.min, respectively. The results of qPCR and iTRAQ demonstrated the much higher expression level of As-SesTPS gene in agarwood than that in whitewood. This study provides a foundation for elucidating the mechanism of agarwood formation in A. sinensis and the potential of the novel gene for improving the quality of artificial agarwood., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Agarwood is a resinous material formed in wounded Aquilaria sinensis in China, which is widely used as an effective traditional Chinese medicine (TCM). This study is aimed to use gas chromatography-mass spectrometry combined with chemometric methods to create reliable criteria for accurate identification of natural agarwood and artificial agarwood, as well as for quality evaluation of artificial agarwood. Natural agarwood and artificial agarwood (stimulated by formic acid or formic acid plus fungal inoculation) were used as standards and controls for the gas chromatography-mass spectrometry (GC-MS) and multivariate analysis. The identification criteria developed were applied to commercial agarwood. A reliable criteria including correlation coefficient of GC-MS fingerprint of natural agarwood and 22 markers of metabolism in natural and artificial agarwood was constructed. Compared with chemically stimulated agarwood (formic acid) and in terms of the 22 markers, artificial agarwood obtained by formic acid stimulation and fungal inoculation were much closer to natural agarwood. The study demonstrates that the chemical components of artificial agarwood obtained by comprehensive stimulated method (formic acid plus fungal inoculation) are much closer to the natural agarwood than those obtained by chemically stimulated method (formic acid), as times goes by. A reliable criteria containing correlation coefficient of GC-MS fingerprint of natural agarwood and 22 metabolism markers can be used to evaluate the quality of the agarwood. As an application case, three samples were identified as natural agarwood from the 25 commercial agarwood by using the evaluation method., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Depression is a common complex psychiatric disorder but its pathophysiological mechanism is not yet fully understood. Metabonomics by GC-MS and multivariate statistical analysis were used to select potential biomarkers associated with CUMS (chronic unpredictable mild stress) depression. The dynamic metabolic changes in rat serum were investigated to find potential disease biomarkers and to investigate the pathology of depression induced by the CUMS depression model. The changes in behavior and serum metabolic profiles were investigated during a three-week CUMS exposure. Serum samples were collected on days 0, 6, 9, 12, 15 and 21, and the serum metabolic profiling was carried out using GC-MS, followed by multivariate analysis. The potential biomarkers were screened from metabolites by principal component analysis and correlation analysis. The peak area of potential biomarkers was used to identify changes in depression in rats and describe their dynamics. Exposure to CUMS for three weeks caused depression-like behavior in rats, as indicated by significant decreases in weight gain, sucrose consumption, ambulation number and rearing numbers. Six potential biomarkers in serum, including glycine (Gly), glutamic acid (Glu), fructose, citric acid, glucose and hexadecanoic acid, were subjected to screening by metabonomics and multivariate statistical analysis. It was found that fructose, glucose and Gly were increased in the model group, while hexadecanoic acid, Glu and citric acid were reduced in the model group. According to the results of principal component analysis and correlation analysis, the correlation coefficient between the behavior scores and potential biomarkers in serum were all more than 0.9. This result suggests that the progression of depression may be associated with perturbation of glycometabolism, amino acid metabolism and energy metabolism. Gly, Glu, fructose, citric acid, glucose and hexadecanoic acid appear to be suitable quantitative diagnostic biomarkers for depression. The representative and unique nature of these biomarkers needs to be verified by pharmacological experiments, including molecular pharmacology investigations of enzymes or genes., (Copyright © 2013 Elsevier Inc. All rights reserved.)

In many cases nanostructures present forbidden spots in their electron diffraction patterns when they are observed by transmission electron microscopy (TEM). To interpret their TEM and high resolution transmission electron microscopy (HRTEM) images properly, an understanding of the origin of these spots is necessary. In this work we comment on the origin of the forbidden spots observed in the [111] and [112] electron diffraction patterns of flat gold triangular nanoparticles. The forbidden spots were successfully indexed as corresponding to the first laue Zone (FOLZ) and the HRTEM images presented a contrast produced by the interference of the zero-order Laue zone (ZOLZ) and FOLZ spots. We discuss the use of the forbidden spots in the study of the structure of nanoparticles and show that they are related to the shape and incompleteness of layers in the very thin particles.

Semiconductor nanocrystals, which have unique optical and electronic properties, have potential for applications in the emerging field of nanoelectronics. To produce nanocrystals cheaply and efficiently, biological methods of synthesis are being explored. We found that E. coli, when incubated with cadmium chloride and sodium sulfide, have the capacity to synthesize intracellular cadmium sulfide (CdS) nanocrystals. The nanocrystals are composed of a wurtzite crystal phase with a size distribution of 2-5 nm. Nanocrystal biosynthesis increased about 20-fold in E. coli cells grown to stationary phase compared to late logarithmic phase. Our results highlight how different genetic and physiological parameters can enhance the formation of nanocrystals within bacterial cells.