The Effect of Chinese Herbal Medicine Formula mKG on Allergic Asthma by Regulating Lung and Plasma Metabolic Alternations.

Asthma is a chronic inflammatory disorder of the airway and is characterized by airway remodeling, hyperresponsiveness, and shortness of breath. Modified Kushen Gancao Formula (mKG), derived from traditional Chinese herbal medicines (TCM), has been demonstrated to have good therapeutic effects on experimental allergic asthma. However, its anti-asthma mechanism remains currently unknown. In the present work, metabolomics studies of biochemical changes in the lung tissue and plasma of ovalbumin (OVA)-induced allergic asthma mice with mKG treatment were performed using ultra high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Partial least squares-discriminate analysis (PLS-DA) indicated that the metabolic perturbation induced by OVA was reduced after mKG treatment. A total of twenty-four metabolites involved in seven metabolic pathways were identified as potential biomarkers in the development of allergic asthma. Among them, myristic acid (L3 or P2), sphinganine (L6 or P4), and lysoPC(15:0) (L12 or P16) were detected both in lung tissue and plasma. Additionally, l-acetylcarnitine (L1), thromboxane B2 (L2), 10-HDoHE (L10), and 5-HETE (L11) were first reported to be potential biomarkers associated with allergic asthma. The treatment of mKG mediated all of those potential biomarkers except lysoPC(15:0) (P16). The anti-asthma mechanism of mKG can be achieved through the comprehensive regulation of multiple perturbed biomarkers and metabolic pathways.

Metabolic responses to Lactobacillus plantarum contamination or bacteriophage treatment in Saccharomyces cerevisiae using a GC-MS-based metabolomics approach.

Bacteriophage can be used as a potential alternative agent for controlling Lactobacillus plantarum contamination during bioethanol production. However, how Saccharomyces cerevisiae respond against contaminative L. plantarum or added bacteriophage remains to be fully understood. In this study, gas chromatography-mass spectrometry and a multivariate analysis were employed to investigate the intracellular biochemical changes in S. cerevisiae cells that were elicited by L. plantarum contamination or bacteriophage treatment. The intracellular metabolite profiles originating from different groups were unique and could be distinguished with the aid of principal component analysis. Moreover, partial least-squares-discriminant analysis revealed a group classification and pairwise discrimination, and 13 differential metabolites with variable importance in the projection value greater than 1 were identified. The metabolic relevance of these compounds in the response of S. cerevisiae to L. plantarum contamination or bacteriophage treatment was discussed. Besides generating lactic acid and competing for nutrients or living space, L. plantarum contamination might also inhibit the growth of S. cerevisiae through regulating the glycolysis in S. cerevisiae. Moreover, increased concentrations of monounsaturated fatty acids secondary to bacteriophage treatment might lead to more membrane fluidity and promote the cell viability of S. cerevisiae.

Rupestonic acids B-G, NO inhibitory sesquiterpenoids from Artemisia rupestris.

Six new guaiane sesquiterpenoids, rupestonic acids B-G (1-6), have been isolated from the whole plants of Artemisia rupestris together with six known compounds (7-12). The structures of the new isolates (1-6) were elucidated on the basis of extensive 1D and 2D NMR analysis, and the absolute configurations were established by electronic circular dichroism (ECD) in combination with density functional theory calculations. In in vitro bioassays, compounds 2 and 6 exhibited significant inhibitory effects on LPS-stimulated NO production in BV-2 microglial cells with IC50 values of 2.6 and 2.2 µM, respectively.

[Chemical constituents from ethyl acetate extract of Artemisia rupestris].

Chemical constituents of ethyl acetate extract of Artemisia rupestris were isolated and purified by various chromatographic methods, including silica gel, ODS and Sephadex LH-20. Their structures were elucidated on the basis of spectroscopic data analysis. 12 compounds were separated from A. rupestris and their structures were identified as 5,4'-dihydroxy -3,6,7-trimethoxy flavone (1), R-(-) -vestitol (2), tricin (3), chrysoeriol (4), 3-indole carboxylic acid (5), esculetin (6), apigenin (7), luteolin (8), trans-caffeic acid (9), casticin (10), chrysosptertin B (11) and artemetin (12). Compound 2 was obtained from the genus Artemisia for the first time, and compounds 1-6 and 9 were separated from this plant for the first time.

Characterization of cupric glutamate extinguishing mechanism of Alexandrium sp. LC3 with two-dimensional electrophoresis and MALDI-TOF MS.

Mechanisms by which cupric glutamate, a novel algicide, extinguishes Alexandrium sp. LC3 are shown in this study. We show that cupric glutamate not only stimulated the production of malonaldehyde (MDA) and dramatically promoted cell plasma membrane permeability (p < 0.01) but also remarkably reduced sulfhydryl (SH) group content (p < 0.01). Analysis of protein expression profiles by two-dimensional electrophoresis (2-DE) indicated that only 47 protein spots were detected in both control and cupric glutamate treated cells. Three reliable spots were identified by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) as ribulose-bisphosphate carboxylase large subunit precursor, RNA polymerase beta chain, and hypothetical protein, which can be well correlated with cupric glutamate stress. Based on above results, we hypothesize that the extinguishing mechanisms include (1) the cell membrane being damaged by cupric glutamate; (2) cupric glutamate probably induced denaturation and disintegration of intracellular protein, which led to inhibition of cell growth.

Determination of the big head carp myofibrillar (Aristichthys nobilis) adenosine triphosphatase activity by ion chromatography.

A method for the determination of adenosine triphosphatase (ATPase) activity of myofibrils of big head carp by using ion chromatography was introduced. Adenosine triphosphate (ATP), adenosine diphosphate (ADP) and orthophosphate (Pi) were separated completely. Recoveries for ATP, ADP and Pi were 98+/-5%, 97+/-4% and 98+/-5%, respectively. Pi liberated from ATP during reaction was monitored by ion chromatography using the suggested method. This method was applicable to the determination of myofibrils ATPase activity for quick quality evaluation of surimi.

Purification and characterization of trimethylamine-N-oxide demethylase from jumbo squid (Dosidicus gigas).

Trimethylamine-N-oxide demethylase (TMAOase) was purified from Jumbo squid (Dosidicus gigas) and characterized in detail herein. The TMAOase was extracted from squid with 20 mM Tris-acetate buffer (pH 7.0) containing 1.0 M NaCl, followed by acid treatment and heat treatment. Then it was purified by deithylaminoethyl-cellulose and Sephacryl S-300 chromatography, subsequently resulting in an 839-fold purification. The molecular mass of the TMAOase was defined to be 17.5 kDa. The optimum pH of the purified TMAOase was 7.0, and its optimum temperature was confirmed to be 55 degrees C. The TMAOase was stable to heat treatment up to 50 degrees C and stable at pH 7.0-9.0. Reducing agents such as DTT, Na2SO3, and NADH were effective at activating TMAOase, and ethylenediaminetetraacetic acid, as well as Mg2+ and Ca2+, could also enhance the activity of TMAOase remarkably, whereas the TMAOase could be significantly inhibited by tea polyphenol, phytic acid and acetic acid. In addition, the TMAOase converted TMAO to dimethylamine and formaldehyde stoichiometrically with a K(m) of 26.2 mM.

Promotion of hexadecyltrimethyleamine bromide to the damage of Alexandrium sp. LC3 by cupric glutamate.

The effect of hexadecyltrimethyleamine bromide (HDTMAB) on the removal of Alexandrium sp. LC3 under cupric glutamate stress was investigated. Toxic effect of cupric glutamate on A lexandrium sp. LC3 was significantly promoted in the presence of HDTMAB, especially at 3.0 cmc of HDTMAB. It was found that the sulfhydryl group content of the cell decreased, while the malonaldehyde content and membrane permeability increased when Alexandrium sp. LC3 was treated with HDTMAB and cupric glutamate complex, compared with cupric glutamate alone. The data suggest that HDTMAB might stimulate the damage of Alexandrium sp. LC3 by enhancing the membrane permeability.

Aberrant purine metabolism in allergic asthma revealed by plasma metabolomics.

Asthma is a disease characterized by chronic relapsing airways, and its etiology remains incompletely understood. To better understand the metabolic phenotypes of asthma, we investigated a plasma metabolic signature associated with allergic asthma in ovalbumin (OVA)-sensitized mice by using ultra high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Sixteen metabolites were characterized as potential pathological biomarkers related to asthma. Among them, 6 (dodecanoic acid (P1), myristic acid (P2), phytosphingosine (P3), sphinganine (P4), inosine (P13) and taurocholic acid (P15)) were first reported to have potential relevance in the pathogenesis of experimental asthma. The identified potential biomarkers were involved in 6 metabolic pathways and achieved the most entire metabolome contributing to the formation of allergic asthma. Purine metabolism was the most prominently influenced in OVA-induced asthma mice according to the metabolic pathway analysis (MetPA), suggesting that significantly changes in inflammatory responses in the pathophysiologic process of asthma. The metabolites of purine metabolism, especially uric acid (P12) and inosine (P13), may denote their potential as targeted biomarkers related to experimental asthma. The decreased plasma uric acid (P12) suggested that inflammation responses of allergic asthma inhibited the activity of xanthine oxidase in purine metabolism, and manifested the severity of asthma exacerbation. The increased level of inosine (P13) suggests that inflammatory cells induce adenosine triphosphate (ATP) breakdown, resulting in excessive expression of adenosine deaminase (ADA) in the formation of allergic asthma. These findings provided a novel perspective on the metabolites signatures related to allergic asthma, which provided us with new insights into the pathogenesis of asthma, and the discovery of targets for clinical diagnosis and treatment.

Effects of exogenous Ca2+ on the membrane permeability, MDA and SH group content of Alexandrium sp. LC3 under surfactant stress.

The effect of Ca2+ on the removal of Alexandrium sp. LC3 under HDTMAB stress was investigated. The results showed that the toxic effect of HDTMAB on Alexandrium sp. LC3 was significantly reduced in the presence of Ca2 , especially under 4 mmol/L of Ca2+. To understand the underlying mechanism, the SH group and MDA content of the cell membrane and membrane permeability were measured. It was found that the SH content of cell member increased, the MDA content and membrane permeability decreased when Alexandrium sp. was treated with Ca2+ and HDTMAB complex, compared with using HDTMAB only. The data suggested that Ca2+ might promote HDTMAB stress resistance of Alexandrium sp. LC3 by reducing the permeability and increasing the stability of cell membrane.

Metabolic changes during the pu-erh tea pile-fermentation revealed by a liquid chromatography tandem mass-spectrometry-based metabolomics approach.

In the current study, liquid chromatography-mass spectrometry combined with multivariate statistical analyses was employed to investigate the time-varying biochemical changes during the pile-fermentation process with the emphasis on the active ingredients to clarify the manufacturing process of ripened pu-erh tea as a whole. The metabolite profiles of different manufacturing processes were unique and could be distinguished with the aid of principal component analysis. Furthermore, partial least-squares discriminant analysis revealed a pairwise discrimination between the raw material group and pile-fermentation process groups or the final product group, and 48 differential metabolites with variable importance in the projection value greater than 1 were identified, which was confirmed by the subsequent hierarchical cluster analysis. These results highlight our current understanding of the exact changing process of the bioactive compounds during the pile fermentation, and the global change of these bioactive compounds provides the special flavor, taste, and health promoting effects of ripened pu-erh tea.

SURFACTANT PROMOTION OF THE INHIBITORY EFFECTS OF CUPRIC GLUTAMATE ON THE DINOFLAGELLATE ALEXANDRIUM(1).

We studied cupric glutamate as a novel algicide for marine harmful algae and hexadecyltrimethyleamine bromide (HDTMAB) as an accelerant. Cupric glutamate had an excellent ability to inhibit the growth of Alexandrium sp. LC3, but the inhibition efficiency did not increase with higher cupric glutamate concentration. The studies on the inhibition ofAlexandrium sp. LC3 by cupric sulfate or cupric glutamate showed that cupric glutamate had a higher inhibition rate than cupric sulfate (P < 0.05). HDTMAB could significantly enhance the inhibition by cupric glutamate (P < 0.05). Ultrastructural changes of Alexandrium sp. LC3 under cupric sulfate, cupric glutamate, and cupric glutamate-HDTMAB combined treatment were studied with TEM. Under these stresses, the integrity of the cell plasma membranes (cell plasma membrane, chloroplast and mitochondria membranes) was destroyed. The degree of damage under cupric glutamate-HDTMAB combined treatment was more severe than under the other stresses. These results indicated that mechanistically cupric glutamate inhibits algal growth by destroying the cell membranes, and that HDTMAB promotes this process, which induced mass extravasation of intracellular components and more copper ion entry into the plasma.