Screening of Human Gut Bacterial Culture Collection Identifies Species That Biotransform Quercetin into Metabolites with Anticancer Properties.

We previously demonstrated that flavonoid metabolites inhibit cancer cell proliferation through both CDK-dependent and -independent mechanisms. The existing evidence suggests that gut microbiota is capable of flavonoid biotransformation to generate bioactive metabolites including 2,4,6-trihydroxybenzoic acid (2,4,6-THBA), 3,4-dihydroxybenzoic acid (3,4-DHBA), 3,4,5-trihyroxybenzoic acid (3,4,5-THBA) and 3,4-dihydroxyphenylacetic acid (DOPAC). In this study, we screened 94 human gut bacterial species for their ability to biotransform flavonoid quercetin into different metabolites. We demonstrated that five of these species were able to degrade quercetin including Bacillus glycinifermentans, Flavonifractor plautii, Bacteroides eggerthii, Olsenella scatoligenes and Eubacterium eligens. Additional studies showed that B. glycinifermentans could generate 2,4,6-THBA and 3,4-DHBA from quercetin while F. plautii generates DOPAC. In addition to the differences in the metabolites produced, we also observed that the kinetics of quercetin degradation was different between B. glycinifermentans and F. plautii, suggesting that the pathways of degradation are likely different between these strains. Similar to the antiproliferative effects of 2,4,6-THBA and 3,4-DHBA demonstrated previously, DOPAC also inhibited colony formation ex vivo in the HCT-116 colon cancer cell line. Consistent with this, the bacterial culture supernatant of F. plautii also inhibited colony formation in this cell line. Thus, as F. plautii and B. glycinifermentans generate metabolites possessing antiproliferative activity, we suggest that these strains have the potential to be developed into probiotics to improve human gut health.

Conjugation of antifungal benzoic acid derivatives as a path for detoxification in Penicillium brasilianum, an endophyte from Melia azedarach.

In this study, the consumption of 4-bromobenzoic acid and 4-chlorobenzoic acid by the fungus Penicillium brasilianum, an endophyte from Melia azedarach is evaluated. This fungus metabolizes these halobenzoic acids to produce three new brominated compounds, which have been isolated and characterized, and three new chlorinated derivatives identified by HRMS. Among these products, (4-bromobenzoyl)proline has been also chemically synthesized and employed in biological assays, thus providing insights for the elucidation of the defense mechanism of P. brasilianum towards these halobenzoic acids.

Characterization of synthetic routes to 'Bromo-DragonFLY' and benzodifuranyl isopropylamine homologues utilizing ketone intermediates. Part 1: synthesis of ketone precursors.

Bromo-DragonFLY (BDF) and many of its analogues are misused as recreational drugs due to their potency as psychoactive substances. To date, none of the published routes to these designer amphetamines have exploited a ketone intermediate. It is well known that benzyl methyl ketone (BMK) can be employed as a precursor in the synthesis of amphetamine. Similarly, it is reasonable to assume that ketone precursors may potentially be utilized in the clandestine synthesis of BDF and its homologues. This paper describes the multifaceted synthesis of novel precursor ketones structurally related to BDF, namely benzodifuranyl propanone 16, its tetrahydrobenzodifuranyl homologue 8, and their brominated analogues 12 and 20. Their characterization by Fourier transform infrared spectroscopy (FTIR), proton nuclear magnetic resonance spectroscopy ((1) H-NMR), carbon nuclear magnetic resonance spectroscopy ((13) C-NMR), high performance liquid chromatography (HPLC), gas chromatography (GC) and mass spectrometry (MS) is also described.

Growth and characterization of Methyl 2-amino-5-bromobenzoate crystal for NLO applications.

Good quality single crystal of organic Methyl 2-amino-5-bromobenzoate (M2A5B) was grown using slow evaporation solution growth technique. The grown crystal was confirmed by single crystal X-ray diffraction. The functional groups and vibrational frequencies were identified using FT-IR and FT-Raman spectral analyses. The presence of hydrogen and carbon atoms in the grown sample was confirmed with proton and carbon NMR spectral studies. The optical energy band gap of the title compound is found to be 2.7 eV from the optical transmission spectra. The refractive indices n(x), n(y), and n(z) were found to be 1.569, 1.587 and 1.600, respectively using Brewster's angle method. The melting point of the material obtained with melting point apparatus is 74°C. Thermal stability of the grown crystal was studied by thermogravimetric analysis (TGA) and differential thermal analysis (DTA). The mechanical behaviour of the grown crystal was analyzed with Vicker's microhardness tester. The particle size dependent second harmonic generation efficiency for M2A5B was evaluated by Kurtz-Perry powder method using Nd:YAG laser, which established the existence of phase matching.

2-Ethylhexyl tetrabromobenzoate and bis(2-ethylhexyl) tetrabromophthalate flame retardants in the Great Lakes atmosphere.

Two relatively new flame retardants, 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (TBB) and bis(2-ethylhexyl)-tetrabromophthalate (TBPH), were identified and quantitated in gas and particle-phase air samples collected from six sites near the shores of the Great Lakes. TBB and TBPH were detected in more than half of the samples collected from 2008 to 2010. Urban areas, such as Chicago and Cleveland, showed the highest concentrations (0.36-290 pg/m(3)), while remote areas, such as Eagle Harbor and Sleeping Bear Dunes, exhibited the lowest levels (0.050-32 pg/m(3)). The atmospheric concentrations of TBB and TBPH increased rapidly and significantly over this time period, perhaps indicating that these compounds are replacing the polybrominated diphenyl ethers (PBDEs), which have been removed or soon will be removed from the marketplace.

Expanding the chiral pool: oxidation of meta-bromobenzoic acid by R. eutrophus B9 allows access to new reaction manifolds.

Metabolism of meta-bromobenzoic acid by the blocked mutant Ralstonia eutrophus B9 affords an enantiopure dearomatised halodiene-diol which we demonstrate is a versatile chiron for organic synthesis. The presence of the halogen leads to reactivity that is distinct to that observed for the non-halogenated analogue and also serves as a synthetic handle for further functionalisation.

[Bromo-Dragonfly poisoning of 18-year-old male]. / Bromo-Dragonfly-forgiftning af 18-årig mand.

Bromo-Dragonfly (BDF) is a designer drug chemically related to LSD and amphetamine. The first registration in Denmark was in October 2007. In the present case an 18-year-old man ingested about 2 ml BDF and developed acute psychosis. At the hospital the patient displayed, hyperpyrexia, tachycardia, tachypnea and hypertension. Because of severe hallucinations and agitation the patient was detained at the hospital and received symptomatic treatment with large doses of benzodiazepines. Four days after hospitalization, the patient discharged himself without significant symptoms or paraclinical findings.

Photodegradation pathways of nonabrominated diphenyl ethers, 2-ethylhexyltetrabromobenzoate and di(2-ethylhexyl)tetrabromophthalate: identifying potential markers of photodegradation.

Photodegradation kinetics of several polybrominated diphenyl ethers (PBDEs), particularly decabromodiphenyl ether (BDE 209), have been reported in various matrixes, demonstrating that it photodegrades primarily via debromination. However, it has been difficult to determine the primary pathways by which bromine is cleaved from BDE 209 to form nona- and octabrominated congeners. In this study, photodegradation of the three nonaBDE congeners (i.e., BDE 206, 207, and 208) was examined individually in three different solvents exposed to natural sunlight and then analyzed to identify the primary degradation products. Rapid degradation of nonaBDEs (half-lives ranging from 4.25 to 12.78 min) was observed coincident with formation of octa- and heptabrominated PBDEs. BDE 207 photodegraded most rapidly while BDE 206 photodegraded the slowest. The photodegradation pathways of each nonaBDE congener were consistent among the different solvent matrixes tested; however, mass balances were found to vary with the type of solvent used in the experiment (recovery ranging from 76 to 95%). The octabrominated congener, BDE 202, and the ratio of BDE197 to BDE 201,were identified as congeners that may serve as environmental markers of photolytic debromination of decaBDE. Additional photodegradation studies were conducted with two new brominated flame retardants used in replacements for pentaBDE mixtures: 2-ethylhexyltetrabromobenzoate (TBB) and di(2-ethylhexyl)-tetrabromophthalate (TBPH). Both TBB and TBPH underwent photolysis more slowly than nonaBDEs (half-lives ranging from 85.70 to 220.17 min) and primarily formed debrominated products.

Solid-phase synthesis of isocoumarins: a traceless halocyclization approach.

A straightforward and traceless solid-phase methodology was developed for the synthesis of isocoumarins. This two-step process involves a Sonogashira cross-coupling reaction between polymer-bound 2-bromobenzoates and terminal alkynes, followed by an electrophile-induced halocyclization of the resulting 2-(alk-1-ynyl)benzoates through activation of the triple bond with the subsequent release of the 3-substituted 4-haloisocoumarins. This polymer-bound parallel synthetic approach allowed us to achieve large diversity in good to excellent yields and purities.

A fatal poisoning involving Bromo-Dragonfly.

This paper reports a fatal overdose case involving the potent hallucinogenic drug Bromo-Dragonfly (1-(8-bromobenzo[1,2-b; 4,5-b']difuran-4-yl)-2-aminopropane). In the present case, an 18-year-old woman was found dead after ingestion of a hallucinogenic liquid. A medico-legal autopsy was performed on the deceased, during which liver, blood, urine and vitreous humour were submitted for toxicological examination. Bromo-Dragonfly was identified in the liver blood using UPLC-TOFMS, and was subsequently quantified in femoral blood (0.0047 mg/kg), urine (0.033 mg/kg) and vitreous humour (0.0005 mg/kg) using LC-MS/MS. Calibration standards were prepared from Bromo-Dragonfly isolated from a bottle found next to the deceased. The structure and purity of the isolated compound were unambiguously determined from analysis of UPLC-TOFMS, GC-MS, HPLC-DAD, (1)H and (13)C NMR data and by comparison to literature data. The autopsy findings were non-specific for acute poisoning. However, based on the toxicological findings, the cause of death was determined to be a fatal overdose of Bromo-Dragonfly, as no ethanol and no therapeutics or other drugs of abuse besides Bromo-Dragonfly were detected in the liver, blood or urine samples from the deceased. To our knowledge, this is the first report of quantification of Bromo-Dragonfly in a biological specimen from a deceased person. This case caused the drug to be classified as an illegal drug in Denmark on 5th December 2007.

Palladium-catalyzed one-step synthesis of isoindole-1,3-diones by carbonylative cyclization of o-halobenzoates and primary amines.

The palladium-catalyzed aminocarbonylation of o-halobenzoates produces 2-substituted isoindole-1,3-diones in good yields. This methodology provides a good one-step approach to this important class of heterocycles and tolerates a variety of functional groups, including methoxy, alcohol, ketone, and nitro groups.

Direct analysis of illicit drugs by desorption atmospheric pressure photoionization.

The feasibility of desorption atmospheric pressure photoionization (DAPPI) in the direct analysis of illicit drugs was demonstrated by the analysis of confiscated drug samples of various forms such as tablets, blotter paper, and plant resin and bloom. 3,4-Methylenedioxymethamphetamine (MDMA), amphetamine, phenazepam, and buprenorphine were detected from the analyzed tablets, lysergic acid diethylamide (LSD) and bromobenzodifuranylisopropylamine (bromo-Dragonfly, ABDF) from blotter paper, and Delta(9)-tetrahydrocannabinol (THC) and cannabinol from Cannabis Sativa bloom and resin. The amphetamines, phenazepam and ABDF showed protonated molecules independent of the solvent used, whereas buprenorphine, LSD and the cannabinoids showed molecular ions with toluene and protonated molecules with acetone as the solvent.

Method to determine enantiomeric excess of glucose by nonchiral high-performance liquid chromatography using circular dichroism detection.

Using nonchiral high-performance liquid chromatography (HPLC) coupled with a circular dichroism (CD) detector, we developed a new method to determine the enantiomeric excess (e.e.) of glucose derivatized with chromophores (p-bromobenzoate). This method is suitable for microscale analysis of glycoside and saccharide. As an application of this method, the enantiomeric excess of the component glucose of a glucan was assayed.

Oxidative DNA cleavage by free and oligonucleotide-conjugated O-bromobenzoic acid in the presence of copper(II) ions.

o-Bromobenzoic acid was found to promote copper-dependent reactive oxygen species formation from molecular oxygen, resulting in DNA base modification and backbone cleavage. The oligonucleotide conjugate bearing 5-(4'-aminopropyl-sulfomoyl)-2-bromobenzoic acid as a reactive group was synthesized and DNA cleavage activity of this oligonucleotide conjugate was tested on a model deoxyoligonucleotide.

Antiparallel pleated beta-sheets observed in crystal structures of N,N-bis(trichloroacetyl) and N,N-bis(m-bromobenzoyl) gramicidin S.

Despite intensive efforts, the structures of gramicidin S (GS) [cyclo(-Val-Orn-Leu-d-Phe-Pro-)(2)] and its analogues have not been elucidated by the X-ray diffraction method, except for the GS-urea complex (Hull et al., Nature 275, 206-207, 1978; Tishchenko et al., Acta Cryst. D53, 151-159, 1997). We focused on the acetylation of GS to obtain suitable crystals for X-ray diffraction. The amino groups of Orn residues were capped with trichloroacetic and m-bromobenzoic acids. Both trichloroacetyl and m-bromobenzoyl GSs (TcGS and BzGS, respectively) are hydrophobic and their properties are similar to those of acetyl-GS (AcGS). Although it is well known that AcGS yields hexagonal crystals, TcGS and BzGS yield monoclinic and orthorhombic crystals in aqueous dimethylformamide solution, respectively. Their cell volumes were approximately one-fourth or one-eighth of the hexagonal cell volume. The crystal structures of TcGS and BzGS were determined as the first examples of acetylated GS analogues: TcGS, C(64)H(90)N(12)O(12)Cl(6). 3(C(3)H(7)NO), M(r) = 1651.47, monoclinic, P2(1), a = 15.4366(6) A, b = 18.5312(4) A, c = 16.4774(6) A, beta = 14.160(2) degrees, V = 4300.6(2) A(3), Z = 2; and BzGS, C(64)H(98)N(12)O(12)Br(2). 1.54(H(2)O), M(r) = 1535.21, orthorhombic, P2(1)2(1)2(1), a = 16.748(10) A, b = 18.834(5) A, c = 28.558(10) A, V = 9008(7) A(3), Z = 4. Both these peptide molecules formed an antiparallel pleated beta-sheet, and pseudo twofold symmetries existed in the repeated sequence. beta-Turns formed at the fragments of d-Phe-Pro were classified into type II' based on their characteristics. The peptide conformations of TcGS and BzGS were similar to each other, and these structural features agreed with those of structures proposed by the previous studies.

Syntheses and stereochemical revision of pseudopterosin G-J aglycon and helioporin E.

Revised structures are proposed for pseudopterosin G-J aglycon and helioporin E.

Oligosaccharide microscale analysis by circular dichroic spectroscopy: reference spectra for chromophoric D-fructofuranoside derivatives.

The microscale analytical method that is being developed in this group for the structure determination of oligosaccharides yields monosaccharide derivatives bearing two types of chromophores suitable for exciton-coupling, namely, 4-bromobenzoate (lambda max 245 nm) and 4-methoxycinnamate (lambda max 311 nm). Comparison of the circular dichroic (CD) curves of these subunits to those in the reference library allows for the determination of the sugar identities, linkage positions, and the absolute configurations. The 32 possible derivatives of methyl alpha- and beta-D-fructofuranosides bearing four chromophores were prepared and their CD spectra recorded. These data serve to extend the CD library, which already encompasses pyranoside derivatives with the gluco-, galacto-, and manno-configurations, and extend the utility of this methodology to the analysis of fructose-containing oligosaccharides.