Panax ginseng has anti-infective activity against opportunistic pathogen Pseudomonas aeruginosa by inhibiting quorum sensing, a bacterial communication process critical for establishing infection.

Virulent factors produced by pathogens play an important role in the infectious process, which is regulated by a cell-to-cell communication mechanism called quorum sensing (QS). Pseudomonas aeruginosa is an important opportunistic human pathogen, which causes infections in patients with compromised immune systems and cystic fibrosis. The QS systems of P. aeruginosa use N-acylated homoserine lactone (AHL) as signal molecules. Previously we have demonstrated that Panax ginseng treatment allowed the animals with P. aeruginosa pneumonia to effectively clear the bacterial infection. We postulated that the ability to impact the outcome of infections is partly due to ginseng having direct effect on the production of P. aeruginosa virulence factors. The study explores the effect of ginseng on alginate, protease and AHL production. The effect of ginseng extracts on growth and expression of QS-controlled virulence factors on the prototypic P. aeruginosa PAO1 and its isogenic mucoid variant (PAOmucA22) was determined. Ginseng did not inhibit the growth of the bacteria, enhanced the extracellular protein production and stimulated the production of alginate. However, ginseng suppressed the production of LasA and LasB and down-regulated the synthesis of the AHL molecules. Ginseng has a negative effect on the QS system of P. aeruginosa, may explain the ginseng-dependent bacterial clearance from the animal lungs in vivo in our previous animal study. It is possible that enhancing and repressing activities of ginseng are mutually exclusive as it is a complex mixture, as shown with the HPLC analysis of the hot water extract. Though ginseng is a promising natural synergetic remedy, it is important to isolate and evaluate the ginseng compounds associated with the anti-QS activity.

Electrospray tandem mass spectrometric study of alkyl 1-methylpyridinium ether derivatives of alcohols.

The fragmentation of 15 alkyl 1-methylpyridinium ether derivatives (D+) of primary and secondary alcohols, benzylic alcohols and phenyl-substituted alcohols was investigated using energy-resolved tandem mass spectrometry. Fragmentation pathways and mechanisms, including the influence of substituents, on the formation of the major product ions, which appear at m/z 110, [D - 109]+ and [D - 111]+, were postulated. Comparison of the abundances of these ions in the product ion spectra of isomeric alcohol derivatives, obtained at the same center-of-mass collision energy (2.0 eV), was found to provide the ability to differentiate among some isomers.

Electrospray tandem mass spectrometric study of ferrocene carbamate ester derivatives of saturated primary, secondary and tertiary alcohols.

The fragmentation pathways and mechanisms for 27 ferrocene carbamate esters of saturated alkyl primary, secondary and tertiary alcohols were investigated using energy-resolved electrospray tandem mass spectrometry (ES-MS/MS). The mechanisms that control the formation and abundance of the three product ions common to all the derivatives, which appear at m/z 201, 227 and 245, were elucidated. Plotting the relative abundances of the three product ions versus a range of center-of-mass collision energies provided a graphical representation of the behavior of the fragmentation process that was directly comparable from compound to compound. As a result, it was possible to compare product ion spectra of the different derivatives to distinguish among different alcohol structural types. Straight-chain primary alcohols were easily distinguished from tertiary alcohols. Both of these structural types, including positional isomers, produced product ion spectra that were distinct from those of beta-branched primary alcohols, or acyclic secondary alcohols or cyclic secondary alcohols. The latter three alcohol types display similar product ion spectra and therefore cannot be distinguished from one another on the basis of these spectra alone.

Ferrocene-based electroactive derivatizing reagents for the rapid selective screening of alcohols and phenols in natural product mixtures using electrospray-tandem mass spectrometry.

The alcohols and phenols of oil of cloves, lemon oil, rose absolut, and oil of peppermint were derivatized with ferroceneoyl azide to generate their ferroceneoyl carbamates. These derivatives are selectively detected at the attomole level, in nanomolar concentrations by electrospray-tandem mass spectrometry (ES-MS/MS) without the need for sample cleanup. The ES-MS/MS analyses of the four essential oils revealed all the expected alcohols, and, in the case of lemon oil, it detected alpha-terpineol as a trace component that was not readily observed by GC-MS. The ES-MS/MS analyses complements the more conventional GC-MS analysis. The ES-MS method has the advantage of speed, selectivity, and sensitivity over GC-MS for detection of a targeted alcohol of a specific mass or structural type. The ES-MS method does not require a chromatographic separation of the components to accomplish its task. In contrast, GC-MS remains the preferred method for the determination of the total constituents of an oil. The ES-MS method may produce artifact ions, especially if the sample is wet and an excess of the ferroceneoyl azide is used; however, the artifacts did not interfere with the analyses.

Derivatization for electrospray ionization mass spectrometry. 3. Electrochemically ionizable derivatives.

In this paper, the use of ferrocene-based "electrochemically ionizable" derivatives to enhance ES-MS analysis of simple alcohols, sterols, and phenols is discussed. These derivatives are designed to take advantage of the electrolysis process inherent to operation of the ES ion source for selective ionization. Derivatization procedures, electrochemical character of the derivatives, and the ES-MS operational parameters necessary to maximize electrochemical ionization and to enhance gas-phase detection are presented with reference to ferrocenecarbamate ester derivatives of a variety of alcohol standards, as well as the ferroceneboronate derivative of the diol, pinacol. Tandem mass spectrometric analysis of the derivatives (precursor and product ion spectra) is shown to provide derivative confirmation, enhanced detection, and additional analyte structure information. The utility of this derivatization approach for the selective detection of alcohols in complicated mixtures is demonstrated using a saw palmetto (Serenoa repens) fruit extract known to contain a variety of alcohols at low levels.

Stabilization of luciferase intermediates by fatty amines, amides, and nitriles.

Long-chain aliphatic amides, mono- and diamines, mono- and dialcohols, and nitriles were found to inhibit the bacterial luciferase reaction by binding with an enzyme intermediate (II, the luciferase-bound 4 alpha-flavin hydroperoxide). Inhibition was determined by measuring the decay rates of the inhibitor-intermediate II complex at different inhibitor concentrations. The data fit a model which was used to estimate the KI. At high concentrations, a plot of the decay rate (k) vs 1/[I] produced a straight line; extrapolation of this to 1/[I] = 0 yields an estimate of the decay rate at infinite inhibitor concentration which we defined as the inhibitor-enzyme-substrate stabilization constant, kESI.

Metalloporphyrin phototoxicity.

The phototoxicities of six metalloporphyrin dimethylesters (i.e. cobalt (Co), copper (Cu), manganese (Mn), nickel (Ni), tin (Sn) and zinc (Zn) were investigated. Hemolysis of human erythrocytes and inactivation of two enzymes (acetylcholinesterase and beta-galactosidase) were used to assess the phototoxic efficacy of these metal chelates. Tin protoporphyrin (SnPP), the only porphyrin found to hemolyze erythrocytes at a concentration of 40 microM (radiation dose, 230 kJ m-2), was much less efficient than either free protoporphyrin IX or hematoporphyrin. SnPP completely inactivated beta-galactosidase at concentrations above 15 microM (radiation dose, 75 kJ m-2) and drastically interfered with acetylcholinesterase activity at a concentration of 150 microM (radiation dose, 75 kJ m-2). CoPP, CuPP, MnPP, NiPP and ZnPP were ineffective photohemolytic agents at 40 microM (radiation dose, 230 kJ m-2), but inactivated acetylcholinesterase and beta-galactosidase activity to varying degrees. These results suggest that (i) metal ions reduce the phototoxicity of protoporphyrin IX, (ii) different metal ions reduce the phototoxic activity of protoporphyrin IX to different degrees and (iii) the biological activities of the various metal complexes vary in different assay systems.