Data_Sheet_1_Exploring the diversity, bioactivity of endophytes, and metabolome in Synsepalum dulcificum.docx

Synsepalum dulcificum exhibits high edible and medicinal value; however, there have been no reports on the exploration of its endophyte resources. Here, we conducted analyses encompassing plant metabolomics, microbial diversity, and the biological activities of endophytic metabolites in S. dulcificum. High-throughput sequencing identified 4,913 endophytic fungal amplicon sequence variants (ASVs) and 1,703 endophytic bacterial ASVs from the roots, stems, leaves, flowers, and fruits of S. dulcificum. Fungi were classified into 5 phyla, 24 classes, 75 orders, 170 families, and 313 genera, while bacteria belonged to 21 phyla, 47 classes, 93 orders, 145 families, and 232 genera. Furthermore, there were significant differences in the composition and content of metabolites in different tissues of S. dulcificum. Spearman’s correlation analysis of the differential metabolites and endophytes revealed that the community composition of the endophytes correlated with plant-rich metabolites. The internal transcribed spacer sequences of 105 isolates were determined, and phylogenetic analyses revealed that these fungi were distributed into three phyla (Ascomycota, Basidiomycota, and Mucoromycota) and 20 genera. Moreover, 16S rDNA sequencing of 46 bacteria revealed they were distributed in 16 genera in three phyla: Actinobacteria, Proteobacteria, and Firmicutes. The antimicrobial activities (filter paper method) and antioxidant activity (DPPH and ABTS assays) of crude extracts obtained from 68 fungal and 20 bacterial strains cultured in different media were evaluated. Additionally, the α-glucosidase inhibitory activity of the fungal extracts was examined. The results showed that 88.6% of the strains exhibited antimicrobial activity, 55.7% exhibited antioxidant activity, and 85% of the fungi exhibited α-glucosidase inhibitory activity. The research suggested that the endophytes of S. dulcificum are highly diverse and have the potential to produce bioactive metabolites, providing abundant species resources for developing antibiotics, antioxidants and hypoglycemic drugs.</p

Syntheses and characterizations of two silver(I) coordination polymers constructed from bipyrazole and dicarboxylate ligands

<div><p>Two silver(I) compounds, [Ag(H₂mbpz)(Hchda)]_n (<b>1</b>) and [Ag₂(H₂mbpz)₂(oba)]_n (<b>2</b>) (where H₂mbpz = 3,3′,5,5′-tetramethyl-4,4′-bipyrazole, H₂chda = <i>trans</i>-cyclohexane-dicarboxylic acid and H₂oba = 4,4′-oxy-bis-benzoic acid), have been synthesized and characterized by single-crystal X-ray diffraction analyses. In both cases, the Ag(I) centers are linked by H₂mbpz ligands to form 1-D Ag(I)-H₂mbpz chains; then, the Ag(I)-H₂mbpz chains connect with two neighboring congeners through Ag⋯Ag interactions, forming 2-D supramolecular layers. In <b>1</b>, the dicarboxylate is monodeprotonated and linked to form an anion chain through strong hydrogen bonding interaction. Such anion chains attach on two sides of the supramolecular layers via Ag–O bonds to form a neutral sandwich-like layered network. In <b>2</b>, the dicarboxylate is fully deprotonated and adopts a <i>μ</i>₂-<i>η</i>1:<i>η</i>1 mode to link two adjacent supramolecular layers to form a twofold interpenetrating 3-D supramolecular network. The thermal stability and luminescence of <b>1</b> were also studied.</p></div

Dopant-Assisted Positive Photoionization Ion Mobility Spectrometry Coupled with Time-Resolved Thermal Desorption for On-Site Detection of Triacetone Triperoxide and Hexamethylene Trioxide Diamine in Complex Matrices

Peroxide explosives, such as triacetone triperoxide (TATP) and hexamethylene trioxide diamine (HMTD), were often used in the terrorist attacks due to their easy synthesis from readily starting materials. Therefore, an on-site detection method for TATP and HMTD is urgently needed. Herein, we developed a stand-alone dopant-assisted positive photoionization ion mobility spectrometry (DAPP-IMS) coupled with time-resolved thermal desorption introduction for rapid and sensitive detection of TATP and HMTD in complex matrices, such as white solids, soft drinks, and cosmetics. Acetone was chosen as the optimal dopant for better separation between reactant ion peaks and product ion peaks as well as higher sensitivity, and the limits of detection (LODs) of TATP and HMTD standard samples were 23.3 and 0.2 ng, respectively. Explosives on the sampling swab were thermally desorbed and carried into the ionization region dynamically within 10 s, and the maximum released concentration of TATP or HMTD could be time-resolved from the matrix interference owing to the different volatility. Furthermore, with the combination of the fast response thermal desorber (within 0.8 s) and the quick data acquisition software to DAPP-IMS, two-dimensional data related to drift time (TATP: 6.98 ms, <i>K</i>₀ = 2.05 cm² V^–1 s^–1; HMTD: 9.36 ms, <i>K</i>₀ = 1.53 cm² V^–1 s^–1) and desorption time was obtained for TATP and HMTD, which is beneficial for their identification in complex matrices

M084 exhibits anxiolytic-like effects in mice.

<p>(A) In the light/dark transition test, M084 treatment (10 mg/kg, 2 hrs) resulted in an increase in the number of entries in the light chamber. (B&C) In elevated plus maze (EPM) test, animals treated with M084 had more entries in all areas (B) and spent more time in the open arms and central area and less time in the closed arms (C) as compared to those treated with vehicle. Diazepam, a known antianxiety agent, was used as a positive control (1.5 mg/kg, 2 hrs). Results are shown as mean ± SEM. N = 12/group, * <i>p</i> < 0.05, ** <i>p</i> < 0.01.</p

M084 exhibits antidepressant-like effects in mice.

<p>(A) The chemical structure of M084. (B&C) M084 reduced immobility time in forced swim test (FST, B) and tail suspension test (TST, C). Amitriptyline, a known antidepressant, was used as a positive control (10 mg/kg, 2 hrs). (D-F) In locomotor activity test, mice treated with vehicle and M084 (10 mg/kg, 2 hrs) showed no difference over the average speed (D), distance in central area and total distance traveled (E), and time in central area (F). Results are presented as mean ± SEM. N = 10–12/group, * <i>p</i> < 0.05, ** <i>p</i> < 0.01.</p

M084 treatment enhances BDNF-TrkB signaling in the prefrontal cortex (PFC).

<p>(A&B) mRNA levels of <i>BDNF</i> and <i>c-fos</i> in hippocampus (<b>A</b>) and PFC (B) of vehicle and M084-treated mice. Amitriptyline was used as a control for known antidepressant. (C&D) mRNA levels of <i>BDNF</i> and <i>c-fos</i> in hippocampus (<b>C</b>) and PFC (D) of vehicle and M084-treated mice subjected to CUS. Control animals were treated with vehicle only without CUS. (E&G) Representative western blots for mature BDNF, p-AKT (S473), total AKT, p-ERK1/2 (T202/204), and total ERK1/2 of PFC (E) and hippocampus (G). (F&H) The statistical results of protein levels of mature BDNF (normalized to that of GAPDH), p-AKT (S473)/total AKT, p-ERK1/2 (T202/204)/total ERK1/2 in PFC (F) and hippocampus (H). For all bar graphs, results are normalized to the control group with only the vehicle treatment and shown as mean ± SEM. N = 6/group,* <i>p</i> < 0.05, ** <i>p</i> < 0.01.</p

M084 treatment reverses the CUS-induced pathological behaviors.

<p>(A) M084 (10 mg/kg, 2 hrs) and amitriptyline (10 mg/kg, 7 days) treatment decreased immobility time in the FST of CUS-exposed mice. (B) In novelty suppressed feeding test (NSFT), CUS increased the latency to feed, while M084 treatment reversed such an effect. (C-E) In locomotor activity test, mice subjected CUS displayed moderate changes over the average speed (C), distance traveled in central area and the total distance traveled (D), and time in central area (E). M084 treatment had no effect on these changes induced by CUS. Results are showed as mean ± SEM. N = 12/group, * <i>p</i> < 0.05, ** <i>p</i> < 0.01, *** <i>p</i> < 0.001.</p

Distribution of M084 in serum, brain and CSF of mice.

<p>Serum, brain, and CSF samples were collected at 2 hours after treatment of M084 or vehicle. Concentrations of M084 were determined with LC-MS/MS analysis. (A) M084 can efficiently cross the blood-brain barrier at pharmacologically relevant doses (10 mg/kg, 2 hrs). Results are shown as mean ± SEM. (B) M084 calibration curve. Correlation coefficient = 0.999654, weighting factor = 1/x. (C-F) Representative LC–MS/MS chromatogram of blank brain sample, serum, brain and CSF, respectively.</p

Pyrazolopyrimidines as Potent Stimulators for Transient Receptor Potential Canonical 3/6/7 Channels

Transient receptor potential canonical 3/6/7 (TRPC3/6/7) are highly homologous receptor-operated nonselective cation channels. Despite their physiological significance, very few selective and potent agonists are available for functional examination of these channels. Using a cell-based high throughput screening approach, a lead compound with the pyrazolopyrimidine skeleton was identified as a TRPC6 agonist. Synthetic schemes for the lead and its analogues were established, and structural–activity relationship studies were carried out. A series of potent and direct agonists of TRPC3/6/7 channels were identified, and among them, <b>4m</b>–<b>4p</b> have a potency order of TRPC3 > C7 > C6, with <b>4n</b> being the most potent with an EC₅₀ of <20 nM on TRPC3. Importantly, these compounds exhibited no stimulatory activity on related TRP channels. The potent and selective compounds described here should be suitable for evaluation of the roles of TRPC channels in the physiology and pathogenesis of diseases, including glomerulosclerosis and cancer