Antimicrobial metabolites produced by endophytic fungi associated with the leaves of Vochysia divergens.

Investigation of the endophytic fungi Nigrospora sphaerica, Nigrospora oryzae, and Pseudofusicoccum stromaticum MeOH fractions isolated from the leaves of Vochysia divergens, a medicinal species from the Brazilian Pantanal, led to the identification of five compounds, namely a new compound (1E,8Z)-10,11-dihydroxy-5,5,8-trimethyl-4-oxocycloundeca-1,8-diene-1-carbaldehyde (1) and four known compounds: 5-methylmellein (2), sclerone (3), daldinone A (4), and lasiodiplodin (5). All compounds were identified using spectroscopic methods, and 1 was corroborated with mass spectrometry, while the known compounds were compared with data in the literature. The relative configuration of compound 1 was determined based on theoretical conformational studies as well as the J experimental values between the hydroxymethyne hydrogens. The antimicrobial activity of the compounds was evaluated. Promising results were obtained for compounds 2, 4, and 5 since they inhibited the bacterium Pseudomonas aeruginosa, an opportunistic pathogen, suggesting the potential of these microorganisms as a source of new antibacterial agents.

Metabolomics Approach Expands the Classification of Propolis Samples from Midwest Brazil.

Propolis samples collected from five areas in Mato Grosso do Sul state, Midwest Brazil, comprising portions of the Cerrado, Pantanal, and Atlantic Forest ecosystems, were investigated for metabolomic profiles and evaluated for antioxidant and antitumor potential. Chemical profiles were determined by HPLC-DAD-MS/MS data and evaluated using principal component analysis and hierarchical clustering analysis to discern chemical composition patterns. Based on phytogeographical origin and chemical composition, 20 potential markers were identified and five groups were distinguished: (I) Cerrado/Central, (II) Atlantic Forest/South, (III) Cerrado-Pantanal transition area/Northwest, (IV) Cerrado/North, and (V) Pantanal/West. Drawing on HPLC-DAD-MS/MS and NMR data, 47 compounds were successfully or tentatively identified, including prenylated phenylpropanoids, flavonoids, isoflavonoids, and di- and triterpenoids, among other constituents. Isoflavonoids, typically found in red propolis from Northeast Brazil, are being reported for the first time in a propolis sample from the Midwest. A new prenylated aromatic compound, (E)-3-[4-hydroxy-3-(2-hydroxy-3-methylbut-3-en-1-yl)-5-(3-methylbut-2-en-1-yl)phenyl]propenoic acid, was obtained. Samples in group II exhibited promising antitumor potential against prostate and breast carcinoma cells, as did samples in groups III and IV against the latter cell line. The sample in group I, despite containing the highest amount of total phenolic compounds and being the only sample to exhibit scavenging activity against DPPH, was not the most cytotoxic against the cell lines tested.

Cytotoxic and Apoptotic Activity of Majoranolide from Mezilaurus crassiramea on HL-60 Leukemia Cells.

Majoranolide, a butanolide isolated from the nonpolar fraction of an ethanol extract of Mezilaurus crassiramea (Lauraceae) fruits, is being reported for the first time in this genus and the third time in plants. Structurally identified from 1D and 2D NMR and HRESIMS data, majoranolide proved cytotoxic against cancer cells-MCF-7 and MDA-MB-231 (breast), HT-29 (colon), PC-3 (prostate), 786-0 (renal), and HL-60 (leukemia)-inhibiting growth in HL-60 cells (GI50 = 0.21 µM) and exhibiting higher selectivity for this line than for nonneoplastic NIH/3T3 murine fibroblasts. Effects on the cell cycle, caspase-3 activation, and plasma membrane integrity were evaluated by flow cytometry. Expression of genes related to apoptotic pathways (BAX, BCL2, BIRC5, and CASP8) was investigated using RT-qPCR. At 50 µM, majoranolide induced cell cycle arrest at G1 in 24 h increased the sub-G1 population in 48 h and increased caspase-3 activation in a time-dependent manner. The compound upregulated BAX and CASP8 transcription (proapoptotic genes) and downregulated BIRC5 (antiapoptotic). Loss of plasma membrane integrity in 30% of cells occurred at 48 h, but not at 24 h, characterizing gradual, programmed death. The results suggest that majoranolide cytotoxicity involves apoptosis induction in HL-60 cells, although other mechanisms may contribute to this cell death.

Rubiaceae-Type Cyclopeptides from Galianthe thalictroides.

Two Rubiaceae-type cyclopeptides, 6-O-methylbouvardin (1) and the new cyclopeptide 5ß-hydroxy-RA-III (2), were isolated from the roots of Galianthe thalictroides. Employing the sulforhodamine B assay, compounds 1 and 2 were tested in vitro against three cancer cell lines--786-0 (kidney carcinoma), PC-3 (prostate carcinoma), and HT-29 (colon carcinoma)--and showed GI50 values ranging from 0.06 to 1.80 µg mL(-1). This is the first report on the isolation of Rubiaceae-type cyclopeptides from a genus other than Rubia or Bouvardia.

Cytotoxic Orbitide from the Latex of Croton urucurana.

The bioactive ethyl acetate phase obtained from the latex of Croton urucurana Baillon afforded a novel orbitide (1), [1-9-NαC]-crourorb A1, that proved active against NCI-ADR/RES (ovary, multidrug-resistance phenotype) cells with the same potency as doxorubicin (positive control) and inactive up to the highest concentration tested against nontumor NIH/3T3 cells. The structure elucidation was based on 1D and 2D NMR spectroscopy, further supported by HRESIMS data and by application of Marfey's method for determination of the absolute configuration of its amino acid residues. This is the first orbitide obtained from C. urucurana.

Evaluation of mutagenic and antimicrobial properties of brown propolis essential oil from the Brazilian Cerrado biome.

Biological, and particularly antimicrobial, activities have been demonstrated for the essential oil of propolis samples worlwide, yet their mutagenic effects remain unknown. To correlate antimicrobial effects with mutagenic risks, the present study evaluated the antifungal and antibacterial activities of the essential oil obtained from brown propolis collected from the Cerrado biome in Midwest Brazil (EOP), testing it against nine pathogenic microorganisms. Evaluation of mutagenic potential was based on the somatic mutation and recombination test (SMART) performed on wing cells of standard (ST) and high-bioactivation (HB) crosses of Drosophila melanogaster. EOP was extracted by hydrodistillation, and sesquiterpenes were characterized by GCâ¿¿MS as its major constituents. The crude oil proved active against Cryptococcus neoformans and Enterococcus faecalis, as did two of its major constituents, spathulenol and (E)-nerolidol â¿¿ the latter being also active against Staphylococcus aureus â¿¿ isolated using chromatographic procedures. No significant increase in the number of somatic mutations was observed in the offspring of ST or HB crosses â¿¿ the latter exhibiting enhanced levels of metabolizing enzymes of the cytochrome P450 type â¿¿ treated with 0.05%, 0.1%, and 0.2% EOP. These findings revealed no mutagenic activity of EOP, even when tested against the HB strain, and demonstrated that its antimicrobial activities are not associated with DNA damage induction (investigated with SMART), suggesting the potential of EOP as a natural preservative.

A new cytotoxic ß-carboline alkaloid from Galianthe thalictroides.

A cytotoxicity-guided fractionation of the roots of Galianthe thalictroides afforded a new ß-carboline alkaloid, 1-(hydroxymethyl)-3-(2-hydroxypropan-2-yl)-2-(5-methoxy-9H- ß-carbolin-1-yl)cyclopentanol, which exhibited strong cytotoxic activity against three human cancer cell lines (MCF-7, 786-0, and UACC62). Its structure was established on the basis of 1D- and 2D-NMR spectroscopic techniques supported by HRMS data.

Cytotoxic aporphine alkaloids from Ocotea acutifolia.

Two new aporphinoid alkaloids, (+)-6 S-ocoteine N-oxide and (+)-norocoxylonine, were isolated from the leaves and trunk bark of OCOTEA ACUTIFOLIA (Lauraceae) along with thirteen aporphine analogues, one morphinan alkaloid, and one flavonoid. The aporphine alkaloids (+)-thalicsimidine and (+)-neolitsine are reported for the first time for the genus OCOTEA. The structures of all compounds were established on the basis of 1D- and 2D-NMR spectroscopic techniques, optical rotation and/or mass spectrometry data. The cytotoxic potential of eight of the aporphine alkaloids against four human cancer cell lines (Hep-2, MCF-7, B16-F10 and 786-0) was also evaluated.

Larvicidal activity against Aedes aegypti of some plants native to the West-Central region of Brazil.

A total of 42 ethanolic extracts from 30 different plant species, native to the Pantanal and Cerrado of the West-Central region of Brazil, have been evaluated for their larvicidal activity against Aedes aegypti larvae, the vector of dengue and dengue hemorrhagic fevers. Among the extracts tested, that obtained from the trunk bark of Ocotea velloziana was the most active. Using a bioassay-directed fractionation of this extract, the active constituent was isolated and characterized as the aporphine alkaloid (+)-dicentrine. Its structure was established on the basis of (1)H and (13)C NMR spectra, optical rotation and by comparison with an authentic sample. This is the first report on the larvicidal activity against A. aegypti of this alkaloid. Our results suggest that (+)-dicentrine may be considered as a promising natural mosquito larvicidal agent.

N-nitrosoanonaine and N-nitrosoxylopine, aporphine alkaloids from Duguetia furfuracea.

Two new aporphine alkaloids, N-nitrosoanonaine (1) and N-nitrosoxylopine (2), were isolated from the aerial parts of Duguetia furfuracea, a weed found in several Brazilian regions. Their structures were elucidated by NMR analysis, CHN analysis, CD, IR, and MS data. The single-crystal X-ray structural determination of the structure of 1 was also performed.