Discovery of urea-based pleuromutilin derivatives as potent gram-positive antibacterial agents.

There is an urgent need to discover new antibacterial drugs and provide new treatment options for clinical antimicrobial resistance (AMR) pathogen infections. Inspired by the structural insights from analyzing the co-crystal structure of lefamulin with the ribosomes of S. aureus, a series of novel pleuromutilin derivatives of phenylene sulfide incorporated with urea moiety were designed and synthesized. The structure-activity relationship (SAR) study revealed that derivatives with urea in the meta position of phenylene sulfide had optimal antibacterial activities in vitro. Among them, 21h was the most potent one against Methicillin-resistant Staphylococcus aureus (MRSA) and clinical AMR Gram-positive bacteria with minimum inhibitory concentrations (MICs) in the range of 0.00195-0.250 µg/mL. And it possessed low resistance frequency, prolonged Post-Antibiotic Effect and the capability to overcome lefamulin-induced resistance. Furthermore, 21h exhibited potent antibacterial activity in vivo in both the thigh infection model and trauma infection model, representing a promising lead for the development of new antibiotics against Gram-positive pathogens, especially for AMR bacteria.

Paspalines C-D and Paxillines B-D: New Indole Diterpenoids from Penicillium brefeldianum WZW-F-69.

Five new indole diterpenoids named paspaline C-D (1-2) and paxilline B-D (3-5), as well as eleven known analogues (6-16), were identified from fungus Penicillium brefeldianum strain WZW-F-69, which was isolated from an abalone aquaculture base in Fujian province, China. Their structures were elucidated mainly through 1D- and 2D-NMR spectra analysis and ECD comparison. Compound 1 has a 6/5/5/6/6/8 hexacyclic ring system bearing 2,2-dimethyl-1,3-dioxocane, which is rare in natural products. Compound 2 has an unusual open F-ring structure. The cytotoxic activities against 10 cancer cell lines and antimicrobial activities against model bacteria and fungi of all compounds were assayed. No compound showed antimicrobial activity, but at a concentration of 1 µM, compounds 1 and 6 exhibited the highest inhibition rates of 71.2% and 83.4% against JeKo-1 cells and U2OS cells, respectively.

Pentacyclic Cytochalasins and Their Derivatives from the Endophytic Fungus Phomopsis sp. xz-18.

Eight new cytochalasins 1-8 and ten known analogs 9-18 were isolated from the endophytic fungus Phomopsis sp. xz-18. The planar structures of the cytochalasins were determined by HR-ESI-MS and NMR analysis. Compounds 1, 2, 9 and 10 were 5/6/6/7/5-fused pentacyclic cytochalasins; compounds 3 and 4 had conjugated diene structures in the macrocycle; and compound 6 had a ß,γ-unsaturated ketone. The absolute configuration of 6 was confirmed for the first time by the octant rule. The acid-free purification process proved that the pentacyclic system was a natural biosynthetic product and not an acid-mediated intramolecular cyclized artifact. The new compounds did not exhibit activities against human cancer cell lines in cytotoxicity bioassays or antipathogenic fungal activity, but compounds 1, 3 and 4 showed moderate antibacterial activity in disk diffusion assays.

Discovery and optimization of 4-oxo-2-thioxo-thiazolidinones as NOD-like receptor (NLR) family, pyrin domain-containing protein 3 (NLRP3) inhibitors.

Aberrant activation of NLRP3 inflammasome is present in a subset of acute and chronic inflammatory diseases. The NLRP3 inflammasome has been recognized as an attractive therapeutic target for developing novel and specific anti-inflammatory inhibitors. Cellular structure-activity relationship-guided optimization resulted in the identification of 4-oxo-2-thioxo-thiazolidinone derivative 9 as a selective and direct small-molecule inhibitor of NLRP3 with IC50 of 2.4 µM, possessing favorable ex vivo and in vivo pharmacokinetic properties. Compound 9 may represent a lead for the development of anti-inflammatory therapeutics for treating NLRP3-driven diseases.

Mycoepoxydiene suppresses HeLa cell growth by inhibiting glycolysis and the pentose phosphate pathway.

Upregulation of glycolysis and the pentose phosphate pathway (PPP) is a major characteristic of the metabolic reprogramming of cancer and provides cancer cells with energy and vital metabolites to support their rapid proliferation. Targeting glycolysis and the PPP has emerged as a promising antitumor therapeutic strategy. Marine natural products are attractive sources for anticancer therapeutics, as evidenced by the antitumor drug Yondelis. Mycoepoxydiene (MED) is a natural product isolated from a marine fungus that has shown promising inhibitory efficacy against HeLa cells in vitro. We used a proteomic approach with two-dimensional gel electrophoresis (2-DE) coupled with mass spectrometry to explore the cellular targets of MED and to unravel the molecular mechanisms underlying the antitumor activity of MED in HeLa cells. Our proteomic data showed that triosephosphate isomerase (TPI) and 6-phosphogluconolactonase (PGLS), which participate in glycolysis and the PPP, respectively, were significantly downregulated by MED treatment. Functional studies revealed that the expression levels of several other enzymes involved in glycolysis and the PPP, including hexokinase 2 (HK2), phosphofructokinase 1 (PFKM), aldolase A (ALDOA), enolase 1 (ENO1), lactate dehydrogenase A (LDHA), and glucose-6-phosphate dehydrogenase (G6PD), were also reduced in a dose-dependent manner. Moreover, the LDHA and G6PD enzymatic activities in HeLa cells were inhibited by MED, and overexpression of these downregulated enzymes rescued HeLa cells from the growth inhibition induced by MED. Our data suggest that MED suppresses HeLa cell growth by inhibiting glycolysis and the PPP, which provides a mechanistic basis for the development of new therapeutics against cervical cancer.

Structure Determinants of Lagunamide A for Anticancer Activity and Its Molecular Mechanism of Mitochondrial Apoptosis.

Marine natural products are served as attractive source of anticancer therapeutics, with the great success of "first-in-class" drugs, such as Yondelis, Halaven, and Brentuximab vendotin. Lagunamides A-C from marine cyanobacterium, Lyngbya majuscula, exhibit exquisite growth inhibitory activities against cancer cells. In this study, we have systematically investigated the structure-activity relationships (SARs) of a concise collection of lagunamide A and its analogues constructed by total chemical synthesis against a broad panel of cancer cells derived from various tissues or organs, including A549, HeLa, U2OS, HepG2, BEL-7404, BGC-823, HCT116, MCF-7, HL-60, and A375. The R configuration of lagunamide A at C-39 position was found to be the structure determinant for anticancer activity. Further molecular mechanism study in A549 cells revealed that lagunamide A induced caspase-mediated mitochondrial apoptosis. Accompanied with the dissipation of mitochondrial membrane potential (Δφm) and overproduction of reactive oxygen species (ROS), lagunamide A led to mitochondrial dysfunction and finally caused cell death. Moreover, both anti- and pro-apoptotic B-cell lymphoma 2 (Bcl-2) family proteins participated in lagunamide A-induced mitochondrial apoptosis, especially myeloid cell leukemia-1 (Mcl-1). Overexpression of Mcl-1 partly rescued A549 cells from lagunamide A-induced apoptosis. This study suggests that lagunamide A may exert anticancer property through mitochondrial apoptosis. Together, our findings would provide insightful information for the design of new anticancer drugs derived from lagunamides.

Additional New Cytotoxic Triquinane-Type Sesquiterpenoids Chondrosterins K-M from the Marine Fungus Chondrostereum sp.

By the method of ¹H NMR prescreening and tracing the diagnostic proton signals of the methyl groups, three additional new triquinane-type sesquiterpenoids-chondrosterins K-M (1-3) and the known sesquiterpenoid anhydroarthrosporone (4)-were isolated from the marine fungus Chondrostereum sp. Their structures were elucidated on the basis of MS, 1D, and 2D NMR data. Chondrosterin K is a rare hirsutane sesquiterpenoid, in which a methyl group was migrated from C-2 to C-6 and has a double bond between C-2 and C-3. Compounds 1-3 showed significant cytotoxicities against various cancer cell lines in vitro.

Aspertetranones A-D, Putative Meroterpenoids from the Marine Algal-Associated Fungus Aspergillus sp. ZL0-1b14.

Aspertetranones A-D (1-4), four new highly oxygenated putative rearranged triketide-sesquiterpenoid meroterpenes, were isolated from the marine algal-associated fungus Aspergillus sp. ZL0-1b14. On the basis of a comprehensive spectroscopic analysis, the planar structures of aspertetranones were determined to possess an unusual skeleton in the terpenoid part. The relative and absolute configurations of the aspertetranones were assigned on the basis of NOESY analysis, X-ray crystallography, and circular dichroism spectroscopy. Compounds 1-4 were evaluated for anti-inflammatory activity in LPS-stimulated RAW264.7 macrophages. Aspertetranone D exhibited an inhibitory effect against IL-6 production with 69% inhibition at 40 µM.

Two new spirooxindole alkaloids from rhizosphere strain Streptomyces sp. xzqh-9.

Two new spirooxindole alkaloids spindomycins A (1) and B (2) were isolated from rhizosphere strain Streptomyces sp. xzqh-9. Their structures were elucidated by comprehensive spectroscopic analyses of NMR and MS data. The absolute configurations of 1 and 2 were determined by experimental and theoretical calculation of electronic circular dichroism (ECD). Antitumor, lactate dehydrogenase, and tyrosine kinase inhibitory activities of two compounds were evaluated, while only spindomycin B (2) exhibited weak inhibitory activity against tyrosine kinase Bcr-Abl.

Three new asperentin derivatives from the algicolous fungus Aspergillus sp. F00785.

Three new asperentin-type compounds, 6-O-α-d-ribosylasperentin (1) and 6-O-α-d-ribosyl-8-O-methylasperentin (2) and 5-hydroxyl-6-O-methylasperentin (3), along with asperentin (4) and its known analogues (5-9), were isolated from a halotolerant Aspergillus sp. strain F00785, an endotrophic fungus from marine alga. Their structures were determined using extensive NMR and HRESIMS spectroscopic analysis, including the X-ray crystallographic data for the assignment of the absolute configurations of compound 9. Compound 4 exhibited highly potent inhibitory activity against crop pathogens, Colletotrichum gleosporioides Penz. and Colletotrichum gleosporioides (Penz.) Sacc.

Use of laser speckle contrast imaging to reveal changes in temperature and blood perfusion in the skin of healthy subjects after administration of heated moxa sticks and daiwenjiu ointment.

OBJECTIVE: To investigate the influence of heated moxa sticks ("moxibustion") and Daiwenjiu ointment (DO) on changes in temperature and blood perfusion volume on the skin of the backs of healthy subjects. METHODS: DO was spread on the left side of the body, and the right side of the body was treated with a heated moxa stick. Images denoting blood perfusion and body temperature were collected 7-8 cm lateral to the spinous process of the sixth thoracic vertebra using laser speckle contrast imaging (LSCI). Data obtained from eight-frame images were analyzed and used to calculate the mean blood perfusion volume. Simultaneously, blood-perfusion images were collected from the body surface and used to compare the change in blood flow on the body surface and the actual position of imaging. RESULTS: After moxibustion, a rapid increase in blood perfusion volume and body temperature was noted in the local skin surface. The maximum blood perfusion volume and body temperature was noted at 20 min (P < 0.05). At 80-110 min after the spreading of DO, a gradual increase was noted in blood perfusion volume (P < 0.05) and body temperature (P < 0.05) compared with the baseline level. The maximum blood perfusion volume was at 110 min. CONCLUSION: Using LSCI, these data revealed a rapid and sharp increase in blood perfusion volume and body temperature after treatment with moxibustion, but the respective changes seen in the DO group were gradual and moderate.

Three nona-2,7-dienoic acid derivatives from saltern derived Micromonospora sp. FXY415.

Three new compounds, apocimycin A-C, were identified from a saltern-derived Micromonospora sp. strain FXY415, isolated from Dongshi saltern, Fujian, China. Their planar structures and relative configuration were confirmed mainly by analysis of 1D- and 2D- NMR spectra. Three compounds belong to 4,6,8-trimythyl nona-2,7-dienoic acid derivatives, apocimycin A also has a phenoxazine nucleus. Apocimycin A-C exhibited weak cytotoxic and antimicrobial activities. Our research showed again that microbial communities in extreme environments are a potential resource in looking for new and bioactive led compounds.

Exploring Diversity through Dimerization in Natural Products by a Rational Tandem Mass-Based Molecular Network Strategy.

The step- and atom-efficient dimerization strategy is frequently used in nature to build structural complexity and diversity. We propose the rationale and structural features of the versatile monomers that are responsible for "diversity through dimerization". Using 5-FAM-maleimide combined with a UHPLC-MS/MS-FBMN workflow, we successfully identified a diverse set of dimeric natural products from fungus Panus rudis F01315, in which all four complex 4'5-ring scaffolds are derived from one monomeric epoxyquinol and endowed with functional diversity.

Kribbella amoyensis sp. nov., isolated from rhizosphere soil of a pharmaceutical plant, Typhonium giganteum Engl.

An actinomycete, designated XMU 198(T), was isolated from the rhizosphere soil of a pharmaceutical plant, Typhonium giganteum Engl., collected in Xiamen City, China. 16S rRNA gene sequence analysis showed that the isolate exhibited highest sequence similarities with Kribbella flavida KACC 20148(T), K. karoonensis Q41(T) and K. alba YIM 31975(T) (98.7, 98.4 and 98.2 %, respectively). The chemotaxonomic characteristics further supported the assignment of strain XMU 198(T) to the genus Kribbella: ll-diaminopimelic acid in the cell-wall peptidoglycan; glucose and galactose with minor amounts of ribose as the whole-cell sugars; polar lipids comprising phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol and unidentified phospholipids; a fatty acid profile characterized by the predominance of iso-C(16 : 0), iso-C(14 : 0) and anteiso-C(15 : 0); and MK-9(H(4)) as the main menaquinone. Gyrase subunit B gene (gyrB) sequence analysis showed that the genetic distances between strain XMU 198(T) and all other members of the genus Kribbella were greater than 0.014, the value used as the threshold for species delineation within this genus. A wide range of genotypic and phenotypic characteristics, as well as DNA-DNA relatedness between strain XMU 198(T) and K. flavida DSM 17836(T) (41.18 %), K. karoonensis Q41(T) (38.02 %) and K. alba DSM 15500(T) (50.58 %), distinguished the isolate from its closest phylogenetic neighbours. On the basis of the above data, a novel species of the genus Kribbella, Kribbella amoyensis sp. nov., is proposed. The type strain is XMU 198(T) ( = DSM 24683(T) = NBRC 107914(T)).

Preparation and defect structure analysis of near-stoichiometric lithium tantalate wafers.

A vapour transfer equilibrium (VTE) method has been used to prepare near-stoichiometric lithium tantalate (NSLT) crystals with different Li contents. The NSLT crystals were tested and analyzed by differential thermal analysis (DTA) and X-ray photoelectron spectroscopy (XPS) to investigate the effect of Li content on the Curie temperature and internal defects of NSLT crystals. This study found that when the Li content increased in the NSLT wafer, the binding energy corresponding to the peak of the Ta4f electron layer in the XPS spectrum first decreased and then increased, indicating that the proportion of Ta valence states was different in wafers with different Li contents. From XPS energy spectrum analysis, it can be seen that the lithium tantalate crystal contains Ta5+, Ta4+, Ta3+ and lower-valence Ta. As the Li content increases in the NSLT wafer, Ta4+ disappears and the proportion of Ta5+ decreases initially, follows by a later increase and then subsequent further decrease. However, the change in proportion of Ta3+ and lower-price Ta is completely opposite to that of Ta5+, showing a trend of first rising, then falling and then finally rising again. Moreover, when the Li content is 49.751% in the NSLT wafer, the proportion of Ta5+ reaches a maximum, showing that at this Li concentration the NSLT crystal has a more perfect lattice structure. In this study, we propose a mixed defect model in which polarons coexist with Li vacancies and Ta inversion, explaining the change in Ta valence state in lithium tantalate crystals. This model is more in line with the observed results in this work. The new hybrid defect model and the variation law of Ta valence state with Li concentration proposed in this paper provide a new direction and experimental proof for the defect study of NSLT crystals, and also provide a theoretical basis to explore the Li content at the best physical properties of NSLT crystals.

Algicidal characteristics of novel algicidal compounds, cyclic lipopeptide surfactins from Bacillus tequilensis strain D8, in eliminating Heterosigma akashiwo blooms.

Heterosigma akashiwo blooms have caused severe damage to marine ecosystems, the aquaculture industry and human health worldwide. In this study, Bacillus tequilensis D8 isolated from an H. akashiwo bloom area was found to exert high algicidal activity via extracellular metabolite production. This activity remained stable after exposure to different temperatures and light intensities. Scanning electron microscopy observation and fluorescein diacetate staining indicated that the algicidal substances rapidly destroyed algal plasma membranes and decreased esterase activity. Significant decreases in the maximum photochemical quantum yield and relative electron transfer rate were observed, which indicated photosynthetic membrane destruction. Subsequently, the algicidal compounds were separated and purified by high-performance liquid chromatography and identified as three surfactin homologues by interpreting high-resolution electrospray ionization mass spectrometry and nuclear magnetic resonance spectroscopy data. Among these, surfactin-C13 and surfactin-C14 exhibited strong algicidal activity against three HAB-causing species, namely, H. akashiwo, Skeletonema costatum, and Prorocentrum donghaiense, with 24 h-LC50 values of 1.2-5.31 µg/ml. Surfactin-C15 showed strong algicidal activity against S. costatum and weak algicidal activity against H. akashiwo but little activity against P. donghaiense. The present study illuminates the algicidal characteristics and mechanisms of action of surfactins on H. akashiwo and their potential applicability in controlling harmful algal blooms.

Acute and Subacute Safety Evaluation of Black Tea Extract (Herbt Tea Essences) in Mice.

Theabrownin (TB) is a heterogeneous biomacromolecule, extracted from tea, with many functional groups. Importantly, TB possesses diverse health benefits, such as antitumor activity and blood lipid-lowering effects. Presently, the content of TB in tea extract is relatively low. Here, we obtained a deep-processed black tea extract with a high content of TB (close to 80%), which was named Herbt Tea Essences (HTE). Currently, this study was designed to evaluate the biosafety of high-content TB products on mice. We implemented acute and subacute toxic experiments to assess its safety on organs, the serum biochemical and molecular levels. In the acute exposure study, we found that the median lethal dose (LD50) value of HTE was 21.68 g/kg (21.06-24.70 g/kg, greater than 5 g/kg), suggesting that HTE had a low acute toxicity. In the 28-day subacute exposure study, our results showed that no abnormal effects were observed in the 40 and 400 mg/kg/day HTE-treated groups. However, we observed slight nephrotoxicity in the 4000 mg/kg/day HTE-treated group. The HTE-induced nephrotoxic effect might involve the inflammatory response activation mediated by the nuclear transcription factor kappa-B (NF-κB) signaling pathway. This study would provide valuable data for the TB safety assessment and promote this natural biomacromolecule application in daily drinking.

Influence of Cooling Rate on Crystallization Behavior of Semi-Crystalline Polypropylene: Experiments and Mathematical Modeling.

As crystallization behavior has a great effect on the injection molding process, the flash differential scanning calorimetry (FSC) method was employed to study the influence of cooling rate on the crystallization behavior of a semi-crystalline polypropylene (PP). As the experimental results show, crystallization temperatures (onset crystallization temperature and maximum crystallization temperature) and crystallinity decrease as the cooling rate increases. In addition, the corresponding mathematical models were established to describe the relationship between the crystallization temperatures/crystallinity and the cooling rate. A revised Tait equation was also carried out based on the mathematical models.

Nigericin is effective against multidrug resistant gram-positive bacteria, persisters, and biofilms.

Multidrug-resistant (MDR) bacteria pose a significant clinical threat to human health, but the development of antibiotics cannot meet the urgent need for effective agents, especially those that can kill persisters and biofilms. Here, we reported that nigericin showed potent bactericidal activity against various clinical MDR Gram-positive bacteria, persisters and biofilms, with low frequencies of resistance development. Moreover, nigericin exhibited favorable in vivo efficacy in deep-seated mouse biofilm, murine skin and bloodstream infection models. With Staphylococcus aureus, nigericin disrupted ATP production and electron transport chain; cell death was associated with altered membrane structure and permeability. Obtaining nigericin-resistant/tolerant mutants required multiple rounds of challenge, and, cross-resistance to members of several antimicrobial classes was absent, probably due to distinct nigericin action with the GraSR two-component regulatory system. Thus, our work reveals that nigericin is a promising antibiotic candidate for the treatment of chronic or recurrent infections caused by Gram-positive bacteria.

5-(Hy-droxy-meth-yl)furan-2-carb-oxy-lic acid.

In the title compound, C(6)H(6)O(4), the furan ring is nearly coplanar with the carboxyl group, the maximum atomic deviation being 0.0248 (9) Å. The crystal packing is stabilized by classical O-H⋯O and weak C-H⋯O hydrogen bonding.