High-throughput screening of novel TFEB agonists in protecting against acetaminophen-induced liver injury in mice.

Macroautophagy (referred to as autophagy hereafter) is a major intracellular lysosomal degradation pathway that is responsible for the degradation of misfolded/damaged proteins and organelles. Previous studies showed that autophagy protects against acetaminophen (APAP)-induced injury (AILI) via selective removal of damaged mitochondria and APAP protein adducts. The lysosome is a critical organelle sitting at the end stage of autophagy for autophagic degradation via fusion with autophagosomes. In the present study, we showed that transcription factor EB (TFEB), a master transcription factor for lysosomal biogenesis, was impaired by APAP resulting in decreased lysosomal biogenesis in mouse livers. Genetic loss-of and gain-of function of hepatic TFEB exacerbated or protected against AILI, respectively. Mechanistically, overexpression of TFEB increased clearance of APAP protein adducts and mitochondria biogenesis as well as SQSTM1/p62-dependent non-canonical nuclear factor erythroid 2-related factor 2 (NRF2) activation to protect against AILI. We also performed an unbiased cell-based imaging high-throughput chemical screening on TFEB and identified a group of TFEB agonists. Among these agonists, salinomycin, an anticoccidial and antibacterial agent, activated TFEB and protected against AILI in mice. In conclusion, genetic and pharmacological activating TFEB may be a promising approach for protecting against AILI.

Identification, flavor characteristics and molecular docking of umami taste peptides of Xuanwei ham.

To better understand the palatable properties of Xuanwei ham, the aqueous extract was isolated, analyzed and combin with sensory evaluation. Of umami-tasting activity and umami-enhancing impact, four new peptides (MDAIKKMQ, RKYEEVAR, YVGDEAQSKRG, and VNVDEVGGEALGR) were extracted and identified by ultrafiltration, gel separation, reverse performance liquid chromatography, and nano-LC-MS / MS. Sensory evaluation results showed that all of them had umami activity and enhanced umami taste, among which VNVDEVGGEALGR had the best effect. These peptides' umami taste thresholds ranged from 0.25 to 0.8 mg/mL. The MSG solution's umami taste threshold ranged from 0.125 to 0.5 mg/mL. Molecular docking results showed that the four umami peptides could be embedded into the binding pocket of the T1R3 cavity of the umami taste receptor T1R1/T1R3, wherein the binding sites Asp219, Asp150, and Thr179 may play crucial roles, and Glu222, Asp108, Glu217 and Glu148 play auxiliary roles. Hydrogen bonding and hydrophobic interactions were the most prominent interaction forces. This study helps to clarify the flavor characteristics of Xuanwei ham and could improve new processing technology.

Flame atomic absorption spectrometry combined with surface-modified magnetic mesoporous silica microspheres by polyethyleneimine for enrichment, isolation and determination of Cu2+ in preserved eggs after high-temperature digestion.

A new efficient magnetic solid-phase extractant based on a surface-modified magnetic mesoporous silica microsphere referred as MMSM-PEI was synthesised and used for the enrichment and isolation of copper ions (Cu2+) in preserved eggs. The physicochemical properties and morphology of MMSM-PEI were characterized by X-ray diffraction (XRD) spectroscopy, Fourier transform infrared spectroscopy (FT-IR), vibration sample magnetometry (VSM), scanning electron microscopy (SEM) and thermos-gravimetric analyses (TGA). The concentrations of trace Cu2+ in the preserved egg were determined by flame atomic absorption spectroscopy (FAAS). The effects of important parameters were examined. The most suitable pH values and temperature for adsorbing Cu2+ were 6.5 and 25 °C, respectively. According to the determination of Cu2+ in egg white, egg yolk and the outer coating mixture (TOCM) of preserved eggs, the spiked recovery and RSD were 94.1-103.8% and 0.96-4.35%, respectively. The limit of detection (LOD) and the limit of quantitation (LOQ) were 0.14 mg/kg and 0.46 mg/kg, respectively. The developed method improved the sensitivity and accuracy of FAAS for the determination of Cu2+ and it could be applied to the determination of trace Cu2+ in real samples.

Effective preparation of magnetic molecularly imprinted polymer nanoparticle for the rapid and selective extraction of cyfluthrin from honeysuckle.

Cyfluthrin is a widely used pesticide. In this study, a sensitive and efficient magnetic molecularly imprinted polymer (MMIP) was prepared by surface molecular imprinting, which used functionalized Fe3O4 particles as magnetic cores. Cyfluthrin was extracted and enriched using magnetic molecularly polymer for analyzing pesticide residue of Chinese herbal medicines. The crystal type, microstructure, particle size, saturation magnetization, and characteristic functional groups of the synthesized MMIPs were analyzed by analysis equipment. The results of isothermal adsorption and kinetic adsorption indicated that MMIPs reached adsorption equilibrium at 30 min, with a maximum capacity of 4.9 mg g-1, which had good adsorption performance, while selective adsorption experiments showed that MMIPs had higher affinity for cyfluthrin. Under the optimized conditions, the limit of detection (LOD) and the limit of quantification (LOQ) were 32.987 ng ml-1 and 109.955 ng ml-1, respectively. And linear range (30-3000ng ml-1) of cyfluthrin with correlation coefficient R2=0.9979, and MMIPs were used in honeysuckle, the recoveries were 91.5%∼97.2%, and RSD was 5.35%∼8.32% (n = 3). It is indicated that the magnetic molecularly imprinted polymer can be used as an effective material for the specific separation of cyfluthrin from honeysuckle.

Four new compounds from Neoboletus magnificus.

Four new compounds, compounds 1, 2, 4, 6, along with two known compounds 3, 5, were isolated from the methanol extract of the fruiting body of Neoboletus magnificus. The structures of compounds were elucidated by HRMS and NMR spectroscopic methods. The in vitro anti-inflammatory activity of the isolated compounds was evaluated.

Magnetic molecularly imprinted polymer combined with high performance liquid chromatography for selective extraction and determination of the metabolic content of quercetin in rat plasma.

In this study, a novel magnetic solid phase extraction adsorbent was prepared, namely Fe3O4@SiO2 as the carrier, quercetin as the template molecule, acrylamide (AM) as the functional monomer, ethylene glycol dimethacrylamide (EGDMA) as the crosslinker, 2,2'-azobisisobutyronitrile (AIBN) as the initiator to prepare magnetic molecularly imprinted polymer (MMIPs) with specific adsorption of quercetin by surface molecular imprinting technique. MMIPs were characterized by Fourier transform infrared spectrometer (FT-IR), X-ray diffraction (XRD) and transmission electron microscope (TEM). The binding properties of MMIPs were evaluated by isothermal adsorption, kinetic adsorption and selective adsorption experiments. The results showed that the saturated adsorption capacity and adsorption time were 17.9 µmol/g and 20 min, respectively, and showed high selectivity for quercetin. Under optimized conditions, the limit of detection (LOD) and limit of quantitation (LOQ) were 55.327 ng/mL and 184.413 ng/mL, respectively. The recovery and RSD were 85.34-95.67% and 3.36-6.59, respectively. We successfully combined magnetic solid phase extraction with high performance liquid chromatography (MSPE-HPLC-UV) for the enrichment, separation and detection of quercetin in rat plasma. The results showed that the peak and half-life of quercetin in plasma were 1.5 h (32.310 µg/mL) and 3 h, respectively. The method was simple, rapid and efficient, and could be applied to the separation and detection of quercetin in complex matrices, and also provides a basis for the separation and identification of other natural chemical components.

Draft Genome Sequence of Fungus Clonostachys rosea Strain YKD0085.

Here, we report the draft genome sequence of Clonostachys rosea (strain YKD0085). The functional annotation of C. rosea provides important information related to its ability to produce secondary metabolites. The genome sequence presented here builds the basis for further genome mining.

A Rapid Centrifugation-Assisted Solid-Phase Extraction and Liquid Chromatography Method for Determination of Loureirin A and Loureirin B of Dragon's Blood Capsules in Rat Plasma and Urine After Oral Administration.

A simple, sensitive and rapid centrifugation-assisted solid-phase extraction (SPE) with high-performance liquid chromatography (SPE-HPLC) method was developed for simultaneous determination of the metabolites loureirin A and loureirin B from Dragon's blood in rat plasma and urine. The development of the extraction procedure included optimization of some important extraction phases. After evaluation, the metabolites of Dragon's blood were extracted by centrifugation-assisted SPE and separated by using HPLC. This method showed good linearity (r(2) > 0.99), and in the rat plasma and urine, the recoveries were 93.1 and 95.7% for loureirin A and were 90.1 and 94.2% for loureirin B. The relative standard deviation (RSD) values of intraday and interday precision in rat plasma and urine for loureirin A were <3.84 and 2.01%, respectively. The RSD values of the intraday and interday precision in rat plasma and urine for loureirin B were below 4.25 and 5.83%, respectively. Thus, the established method is suitable for metabolism studies of loureirin A and loureirin B in rat plasma and urine.

Utility of 4-chloro-7-nitrobenzofurazan for the spectrofluorimetric determination of butylated hydroxyanisole and propyl gallate in foodstuffs.

UNLABELLED: A spectrofluorimetric method is presented for the determination of 2 synthetic phenolic antioxidants (SPAs), butylated hydroxyanisole (BHA), and propyl gallate (PG) in foodstuffs. The proposed method is based on the derivatization of SPAs with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in phosphate buffer of pH 9.0 to yield a highly fluorescent brown product. The optimum experimental conditions have been studied carefully. Linear calibration curves were obtained over the concentration range of 0.20 to 40 µg mL⁻¹ for BHA, and 0.80 to 50 µg mL⁻¹ for PG, using NBD-Cl reagent. The detection limits were 18 ng mL⁻¹ for BHA, 55 ng mL⁻¹ for PG. Intra-day and inter-day relative standard deviations at 3 different concentrations were determined. The high recovery values indicate the accuracy of the proposed methods, and low relative standard deviation values indicate good precision. The results presented in this report show that the applied spectrofluorimetric method is acceptable for the determination of the 2 SPAs in the foodstuffs. Other SPAs, tertiary butyl hydroquinone and butylated hydroxytoluene in foodstuffs do not interfere with the proposed method. PRACTICAL APPLICATIONS: In this spectrofluorimetric method, NBD-Cl as a derivation agent is used to detect synthetic phenolic antioxidants. The method specificity has been greatly improved; there was no interference from other commonly used phenolic substances.

Three new indole alkaloids from Trigonostemon lii.

Three unprecedented indole alkaloids, trigonoliimines A-C (1-3) with a unique polycyclic system, were isolated from the leaves of Trigonostemon lii Y. T. Chang. The structures of 1-3 were determined by the spectroscopic, computational, and CD exciton chirality approaches. Trigonoliimine A showed modest anti-HIV-1 activity (EC(50) = 0.95 microg/mL, TI = 7.9).

Carboline alkaloids from Trigonostemon lii.

Three types with six new carboline alkaloids, trigonostemonines A-F (1- 6), were isolated from the roots and stems of Trigonostemon lii. Their structures were elucidated by spectroscopic analyses. It is the first time that beta-carboline alkaloids have been reported in Trigonostemon species. Trigonostemonine F (6) exhibited moderate cytotoxic activity against HL-60 with an IC (50) value of 16 microM.

Two new norbisabolane sesquiterpenoid glycosides from Glochidion coccineum.

Two new norbisabolane sesquiterpenoid glycosides, glochicoccinosides A (1) and B (2), together with two known compounds, have been isolated from the rhizomes of Glochidion coccineum. Their structures were elucidated by the combination of 1D NMR, 2D NMR, and MS spectral analysis, as well as chemical evidence. Cytotoxic activities and the antioxidant effect of these compounds were evaluated, but none of them showed activity.

Tarennane and tarennone, two novel chalcone constituents from Tarenna attenuata.

Two novel chalcone constituents, tarennane (1) and tarennone (2), together with nine known compounds, were isolated from the whole plant of Tarenna attenuata. By analysis of physical and spectroscopic data, the structures of new compounds were elucidated as (E)-4-[3-(4-hydroxy-3-methoxyphenyl)-acryl]-3,4,5-trimethoxycyclohexa-2,5-dienone (1) and 1,2-bis(3,5-dimethoxy-4-hydroxyphenyl)-3-hydroxypropan-1-one (2). These two compounds were tested for antioxidant activities in the MTT and DPPH assays. Compound 1 revealed potent antioxidant activities against H2O2-induced impairment in PC12 cells, but neither of them showed DPPH radical-scavenging activity with IC50 values of 181.1 and 210.3 microM, respectively.