Microascones, Decahydrofluorene-Class Alkaloids from the Marine-Derived Fungus Microascus sp. SCSIO 41821.

Eight new decahydrofluorene-class alkaloids, microascones A and B (1 and 2), 2,3-epoxyphomapyrrolidone C (3), 14,16-epiascomylactam B (4), 24-hydroxyphomapyrrolidone A (5), and microascones C-E (6-8), along with five known analogs (9-13) were isolated from the marine-derived fungus Microascus sp. SCSIO 41821. Compounds 1 and 2 have an unprecedented complex macrocyclic alkaloid skeleton with a 6/5/6/5/6/5/13 polycyclic system. Their structures and absolute configurations were determined by spectroscopic analysis, quantum chemical calculations of ECD spectra, and 13C NMR chemical shifts. Compounds 10-13 showed selective enzyme inhibitory activity against PTPSig, PTP1B, and CDC25B, and 4, 9, and 10 exhibited strong antibacterial activity against seven tested pathogens. Their structure-bioactivity relationship was discussed, and a plausible biosynthetic pathway for 1-8 was also proposed.

A nanocomposite consisting of etched multiwalled carbon nanotubes, amino-modified metal-organic framework UiO-66 and polyaniline for preconcentration of polycyclic aromatic hydrocarbons prior to their determination by HPLC.

A polyaniline composite doped with etched multi-walled carbon nanotubes and UiO-66-NH2 was prepared by electropolymerization. It was used as a sorbent to extract the polycyclic aromatic hydrocarbons (PAHs) phenanthrene, fluoranthene and pyrene. Its surface morphology, crystal structure and capability of adsorbing PAHs were characterized by scanning electron microscopy, X-ray photoelectron spectrometry, Fourier transform infrared spectrometry and zeta potentiometry. The π stacking and anion-π interactions are shown to play dominant roles in the sorption mechanism. Coupled with high performance liquid chromatography, the composite-modified fiber was applied to detect PAHs in lake water samples by direct immersion extraction. The method excels by (a) wide linear range (0.05-20 ng mL-1), (b) low limits of detection (10 pg mL-1), (c) satisfactory recovery from spiked samples (84.7-113.8%), and (d) good reproducibility (relative standard deviations of <6.5%). The method is superior in terms of costs and reproducibility compared to some pretreatment methods with mass spectrometric detection. Graphical abstractSchematic representation for interaction between PANI-etched MWCNT/UiO-66-NH2 and polycyclic aromatic hydrocarbons (phenanthrene, fluoranthene, pyrene).

New Fluorene Derivatives from Dendrobium gibsonii and Their α-Glucosidase Inhibitory Activity.

Two new compounds, dihydrodengibsinin (1) and dendrogibsol (2), were isolated from the whole plant of Dendrobium gibsonii, together with seven known compounds (3-9). The structures of the new compounds were elucidated by their spectroscopic data. All these isolates were evaluated for their α-glucosidase inhibitory activities. Dendrogibsol (2) and lusianthridin (7) showed strong α-glucosidase inhibitory activity when compared with acarbose. An enzyme kinetic study revealed that dendrogibsol (2) is a noncompetitive inhibitor of α-glucosidase.

Protective effect of hypoxylonol C and 4,5,4',5'-tetrahydroxy-1,1'-binaphthyl isolated from Annulohypoxylon annulatum against streptozotocin-induced damage in INS-1 cells.

We evaluated the protective effects of hypoxylonol C and 4,5,4',5'-tetrahydroxy-1,1'-binaphthyl (BNT) isolated from Annulohypoxylon annulatum on pancreatic ß-cell apoptosis, using the ß-cell toxin streptozotocin (STZ). Hypoxylonol C and BNT restored the STZ-induced decrease in INS-1 cell viability in a dose-dependent manner. In addition, treatment of INS-1 cells with 50 µM STZ resulted in an increase in apoptotic cell death, which was observed as annexin V fluorescence intensity. Apoptotic cell death was decreased by co-treatment with 100 µM hypoxylonol C and 100 µM BNT. Similarly, STZ caused a marked increase in the expression of cleaved caspase-8, caspase-3, Bax, and poly (ADP-ribose) polymerase (PARP), as well as a decrease in the expression of B-cell lymphoma 2 (Bcl-2), which was reversed by co-treatment with 100 µM hypoxylonol C and 100 µM BNT. These findings suggest that hypoxylonol C and BNT play an important role in protecting pancreatic ß-cells against apoptotic damage.

Discovery of Stealthin Derivatives and Implication of the Amidotransferase FlsN3 in the Biosynthesis of Nitrogen-Containing Fluostatins.

Diazobenzofluorene-containing atypical angucyclines exhibit promising biological activities. Here we report the inactivation of an amidotransferase-encoding gene flsN3 in Micromonospora rosaria SCSIO N160, a producer of fluostatins. Bioinformatics analysis indicated that FlsN3 was involved in the diazo formation. Chemical investigation of the flsN3-inactivation mutant resulted in the isolation of a variety of angucycline aromatic polyketides, including four racemic aminobenzo[b]fluorenes stealthins D⁻G (9⁻12) harboring a stealthin C-like core skeleton with an acetone or butanone-like side chain. Their structures were elucidated on the basis of nuclear magnetic resonance (NMR) spectroscopic data and X-ray diffraction analysis. A plausible mechanism for the formation of stealthins D⁻G (9⁻12) was proposed. These results suggested a functional role of FlsN3 in the formation/modification of N⁻N bond-containing fluostatins.

Strengthening detoxication impacts of Coprinus comatus on nickel and fluoranthene co-contaminated soil by bacterial inoculation.

To develop an efficient and environmental-friendly approach to detoxicate nickel (Ni) and fluoranthene co-contaminated soil, the combined application of Coprinus comatus (C. comatus) with Serratia sp. FFC5 and/or Enterobacter sp. E2 was investigated. The pot experiment tested the influences of bacterial inoculation on the growth of C. comatus, content of Ni in C. comatus, Ni speciation in soil, fluoranthene dissipation, soil enzymatic activities, bacterial population and community structure. With the inoculation of bacteria, the fresh weights of C. comatus, concentration of Ni in C. comatus and the dissipation rates of fluoranthene were increased by 17.73-29.38%, 68.97-204.97% and 34.84-60.90%, respectively. Notably, results illustrated that the co-inoculation of FFC5 and E2 showed better effect in biomass enhancement, Ni accumulation and fluoranthene dissipation than solitary inoculation. Simultaneously, higher soil enzymatic and microbiological activities suggested that the integrated detoxication method of bacteria and C. comatus could improve soil quality. Therefore, we can infer that bacterial inoculation strengthened detoxication effect of C. comatus in Ni-fluoranthene co-contaminated soil, indicating that the combined application of C. comatus and bacteria can be an efficient alternative for detoxicating Ni and fluoranthene co-contaminated soil.

The synthesis of dendroflorin.

The first synthesis of dendroflorin has been achieved in 10 steps with an overall yield of 5.5%. The key step in the synthesis features the biphenyl structure is built through Suzuki-Miyaura reaction. In addition, the ortho-localization effect induced by aromatic substituent during the bromination of intermediate 8 is also observed and discussed.

Potential of vancomycin for the enantiomeric resolution of FMOC-amino acids by capillary electrophoresis-ion-trap-mass spectrometry.

The potential of the antibiotic vancomycin (VC) as chiral selector for the enantiomeric separation of amino acids by CE-ESI-MS/MS² was investigated for the first time in this work. Derivatization of amino acids with FMOC-Cl was carried out to enable their interaction with VC as well as the formation of precursor ions with larger m/z which were employed in MS² experiments. The partial filling of a coated capillary was employed to avoid the loss in MS sensitivity originated by the introduction of VC in the ionization source. Under optimized conditions, the simultaneous enantiomeric separation and unequivocal identification of 17 amino acids (two of them being nonprotein amino acids) took place in about 20 min with LODs in the micromolar range.

Antiangiogenic activity of hypoxylonol C.

Hypoxylonol C (1), isolated from the inedible mushroom Hypoxylon truncatum, exhibited inhibitory activities against the migration and tube formation of HUVECs. A cDNA microarray analysis was performed to investigate the target of hypoxylonol C (1) in HUVECs, and it was found that the genes related to cell cycle and adhesion were down-regulated. The down-regulation of mRNA levels of cell cycle and adhesion genes was confirmed by real-time RT-PCR. Cell cycle arrest and suppression of adhesion molecule expression might be plausible mechanisms of actions for the antiangiogenic activity of hypoxylonol C (1).

Benzo(b)fluoranthene and benzo(k)fluoranthene removal by microalgae in the presence of triazine herbicides: Matrix solid-phase dispersion and solid-phase extraction (MSPD/SPE) for HPLC-UV analysis of the different culture components.

Over the last decade, human activities in the industrial and agricultural sectors have significantly increased the concentration of persistent and harmful pollutants in aquatic ecosystems. The use of microorganisms is a green strategy for the bio-removal of certain contaminants. However, other pollutants in the same ecosystems can reduce their degrading activity and even affect their survival. Therefore, this study aimed to evaluate the efficiency of benzo(b)fluoranthene (BbF) and benzo(k)fluoranthene (BkF) removal by Selenastrum capricornutum in the presence of triazine herbicides, compounds mainly used in broadleaf weeds. The interest of this work focused on identifying in which of the microalgal components the degrading activity is best evidenced and affected. For this purpose, the use of solid-phase extraction (SPE) and matrix solid-phase dispersion (MSPD) extraction procedures and HPLC-UV analysis allowed the BbF and BkF trace quantification in biomass, liquid medium, and cell lysate separately from cultures exposed to these polycyclic aromatic hydrocarbons (PAHs) alone or with herbicides. The recovery percentages were between 78 and 94 %, good linearity (r2 ≈ 0.99), precision values measured as RSD < 15 %, and limits of detection (LOQs) at levels of ng mL-1 and ng mg-1 were obtained. The individual PAH amounts measured in the components of microalgae cultures show similar removal kinetics (removal percentages: 82-89 %). Likewise, the analysis demonstrated that the removal of PAHs is not affected in the presence of triazine herbicides (atrazine and cyanazine) and with similar removal percentages (79-86 %) compared to those cultures exposed to individual PAHs (74-83 %). These results support the possible real-world applications of PAH removal by extracts from S. capricornutum in aquatic environments contaminated with PAHs and near agriculture areas where triazine herbicides are used.

Determination of artemether and lumefantrine in anti-malarial fixed-dose combination tablets by microemulsion electrokinetic chromatography with short-end injection procedure.

BACKGROUND: Artemether-lumefantrine (AL) combination therapy is now the most used anti-malarial treatment in the world. Quality control of AL formulations is still a major challenge in developing countries. Until now, only liquid chromatographic methods have been reported in the literature for their analysis. Capillary electrophoretic methods, which present various advantages (low price of capillary, low volumes of electrolyte consumption), may be an alternative to liquid chromatography methods. In this paper, a reliable method was developed and validated for the determination of AL in commercial fixed-dose combination tablets commercialized in Côte d'Ivoire. METHODS: Artemether and lumefantrine were determined by microemulsion electrokinetic chromatography using short-end injection procedure. The two analytes were extracted from tablets by acidified methanol. Pyrimethamine was used as internal standard. Separation was carried out in an uncoated fused silica capillary, 30 cm long × 50 µm internal diameter, using an effective length of 10 cm and a microemulsion composed of octane, butanol, sodium dodecyl sulfate and borate buffer as background electrolyte, a - 500 V x cm(-1) electric field and a detection wavelength of 214 nm. RESULTS: Artemether, lumefantrine and pyrimethamine were separated in 6 min. The method was reliable with respect to selectivity towards formulation excipients, linearity of the response function (r2 > 0.998), recovery studies from synthetic tablets (in the range 99-101%), repeatability (relative standard deviation 1-3%, n = 7 analytical procedures). Application to four commercial formulations containing 20/120 mg of AL per tablet gave a content in good agreement with the declared content. However, the electropherogram of one tablet formulation showed the presence of an ingredient which was not declared. CONCLUSION: The developed MEEKC method can be proposed as an alternative method to liquid chromatography for the determination of artemether and lumefantrine in fixed-dose combination tablet formulations.

Combination of solid phase extraction and surface-enhanced Raman spectroscopy for rapid analysis.

A surface-enhanced Raman spectroscopy (SERS)-active extraction column based on propanethiol modified silver dendrites was fabricated. The column, which combines the advantages of solid phase extraction and SERS, may facilitate the development of rapid analysis with high reliability and accuracy. High temporal stability (under a continuous and intensive laser radiation) and excellent repeatability (repeated extraction and elution) were also achieved using this column. As an example, the quantitative analysis of fluoranthene was accomplished in the concentration range of 0.01-100 µg mL(-1) with this column. The extraction process could be accomplished in 10 s and the total time of one sample analysis including extraction, spectral acquisition, elution and intermediate process could be less than 30 s. This method can greatly simplify the sample preparation and reduce the total analysis time.

Insights into lomaiviticin biosynthesis. Isolation and structure elucidation of (-)-homoseongomycin.

The dimeric diazofluorenes known as the lomaiviticins are produced by the marine bacterium Salinispora pacifica DPJ-0019. Investigation of the fermentation broth of DPJ-0019 has yielded the first monomeric benzo[b]fluorene isolated from this species, (-)-homoseongomycin (13). (-)-Homoseongomycin (13) is related to the known natural product seongomycin (10), which is co-produced with the monomeric diazofluorenes known as the kinamycins. We describe the synthesis of the isotopically labeled derivative homoseongomycin-d5 (14), via the intermediacy of the diazofluorene "prelomaiviticin-d5" (12). Our studies establish that (-)-homoseongomycin (13) may be derived from prelomaiviticin (11) and suggest that 13 and 10 are shunt or detoxification metabolites in lomaiviticin and kinamycin biosynthesis, respectively.

Isolation of lomaiviticins C-E, transformation of lomaiviticin C to lomaiviticin A, complete structure elucidation of lomaiviticin A, and structure-activity analyses.

We describe the isolation of (-)-lomaiviticins C-E (6-8), elucidation of the complete absolute and relative stereochemistry of (-)-lomaiviticin A (1), the synthetic conversion of (-)-lomaiviticin C (6) to (-)-lomaiviticin A (1), and the first evidence that the dimeric diazofluorene of (-)-lomaiviticin A (1) plays a defining and critical role in antiproliferative activity.

Resolution of protected silaproline for a gram scale preparation.

Silaproline is an analogue of proline, which exhibits similar conformational properties. Moreover, the presence of dimethylsilyl group confers to silaproline a higher lipophilicity as well as an improved resistance to biodegradation. This report describes the comparison of two routes to obtain Fmoc-(L) Sip-OH on the gram scale using chiral HPLC resolution.

Hypoxylonols C-F, benzo[j]fluoranthenes from Hypoxylon truncatum.

Four novel benzo[j]fluoranthene derivatives, hypoxylonols C (3), D (4), E (5), and F (6), have been isolated from the mushroom Hypoxylon truncatum, together with two known benzo[j]fluoranthene derivatives, hypoxylonols A (1) and B (2). The structures were established by analysis of NMR spectroscopic data and X-ray diffraction data. Compounds 4 and 5 showed antiproliferative activity against HUVECs (human umbilical vein endothelial cells) and HUAECs (human umbilical artery endothelial cells).

Fluostatins I-K from the South China Sea-derived micromonospora rosaria SCSIO N160.

The strain SCSIO N160 was isolated from a South China Sea sediment sample and was characterized as a Micromonospora rosaria species on the basis of its 16S rRNA gene sequence. Three new fluostatins, I-K (1-3), were isolated from the culture of M. rosaria SCSIO N160, together with six known compounds, fluostatins C-F (4-7), rabelomycin (8), and phenanthroviridone (9). The structure of fluostatin D (5) was confirmed by an X-ray crystallographic study. The absolute configuration of 1 and 3 was assigned by electronic circular dichroism calculations. Compounds 8 and 9 exhibited good antimicrobial activities against Staphylococcus aureus ATCC 29213 with MIC values of 1.0 and 0.25 µg/mL, respectively. Compound 9 also exhibited significant in vitro cytotoxic activities toward SF-268 (IC50 0.09 µM) and MCF-7 (IC50 0.17 µM).

Extraction of phenanthrene and fluoranthene from contaminated sand using palm kernel and soybean oils.

Experimental extraction tests are conducted to investigate feasibility of saturated palm kernel oil (PKO) and unsaturated soybean oil (SO) to extract polycyclic aromatic hydrocarbons (PAHs) from contaminated sand. The extraction rates and efficiencies for lowly contaminated (LC) and highly contaminated (HC) sands at temperatures of 30 °C and 70 °C are evaluated using empirical first order kinetic dissolution models. In LC sand, the extraction is dominated by the diffusion of PAHs adsorbed onto particle surfaces and the direct dissolution of PAH phase. In HC sand, a rapid diffusion of PAHs adsorbed onto particle surfaces and a direct dissolution of PAH phase occur followed by a slower diffusion of PAHs entrapped within the pores and micropores. Larger diffusion resistance during HC sand extractions results in an average 10.8% reduction in extraction efficiencies compared to LC sand. Increased temperature generally increases the mass transfer rates and extraction efficiencies. Additionally, the physicochemical properties of both oils and PAHs also determine the extent of PAH extraction into oil.

Enantiomeric separation of FMOC-amino acids by nano-LC and CEC using a new chiral stationary phase, cellulose tris(3-chloro-4-methylphenylcarbamate).

A novel polysaccharide-based chiral stationary phase (CSP), cellulose tris(3-chloro-4-methylphenylcarbamate), also known as Sepapak-2 or Lux Cellulose-2, has been evaluated for the enantiomeric separation of FMOC derivatives of amino acids. After mobile-phase optimization in nano liquid chromatography (nano-LC) the column enabled the enantiomeric separation of 19 out of 23 amino acids tested, indicating the high chiral recognition power of this new CSP. Subsequently, a comparison of the driving force employed (pressure or voltage) was carried out comparing nano-LC and CEC under the same conditions. Better peak efficiencies and resolution were observed by using CEC experiments, which enabled the chiral discrimination of 20 out of 23 amino acids tested. Finally, in order to show the potential of this new CSP, the determination of the content and the enantiomeric purity of the non-protein amino acid citrulline in food supplements was performed. For that purpose, the method was optimized, evaluated and applied to different commercial samples.