Endothelium-Independent Vasodilatory Effects of Isodillapiolglycol Isolated from Ostericum citriodorum.

Ostericum citriodorum is a plant with a native range in China used in herbal medicine for treating angina pectoris. In this study, we investigated the vasodilatory effects of isodillapiolglycol (IDG), which is one of the main ingredients isolated from O. citriodorum ethyl acetate extract, in Sprague-Dawley rat aortic rings, and measured intracellular Ca2+ ([Ca2+]in) using a molecular fluo-3/AM probe. The results show that IDG dose-dependently relaxed endothelium-intact or -denuded aortic rings pre-contracted with noradrenaline (NE) or potassium chloride (KCl), and inhibited CaCl2-induced contraction in high K+ depolarized aortic rings. Tetraethyl ammonium chloride (a Ca2+-activated K+ channel blocker) or verapamil (an L-type Ca2+ channel blocker) significantly reduced the relaxation of IDG in aortic rings pre-contracted with NE. In vascular smooth muscle cells, IDG inhibited the increase in [Ca2+]in stimulated by KCl in Krebs solution; likewise, IDG also attenuated the increase in [Ca2+]in induced by NE or subsequent supplementation of CaCl2. These findings demonstrate that IDG relaxes aortic rings in an endothelium-independent manner by reducing [Ca2+]in, likely through inhibition of the receptor-gated Ca2+ channel and the voltage-dependent Ca2+ channel, and through opening of the Ca2+-activated K+ channel.

Microwave-assisted kinetic resolution of homochiral (Z)-cyclooct-5-ene-1,2-diol and (Z)-2-acetoxycyclooct-4-enyl acetate using lipases.

Over the last decade, the use of biocatalysts has become an attractive alternative to conventional chemical methods, especially for organic synthesis, due to their unusual properties. Among these enzymes, lipases are the most widely used, because they are cheap, easily available, cofactor-free, and have broad substrate specificity. Combined to microwave heating in non-aqueous medium, recent results suggest that irradiation may influence the enzyme activity. This Communication reports the benefits of lipases and the microwave irradiation on the kinetic resolution of racemic homochiral (Z)-cyclooct-5-ene-1,2-diol and (Z)-2-acetoxycyclooct-4-enyl acetate. In order to best achieve the kinetic resolution, different parameters were studied including the type of lipase, the temperature, the impact of microwave power compared to conventional heating. Optimization of the reaction parameters lead to the obtainment of highly enriched or enantiopure diols and diesters in a clean, efficient and safe way.

Isolation, structure determination, and antiaging effects of 2,3-pentanediol from endophytic fungus of Curcuma amada and docking studies.

An endophytic fungus was isolated from the rhizomes of Curcuma amada (Zingiberaceae), which was identified as Fusarium oxysporum on the basis of its morphological and molecular characters. Chromatographic separation and spectroscopic analysis of the fungal metabolite (chloroform extract) led to the identification of one pure compound having molecular formula C5H12O2, i.e., 2,3-pentanediol (1). Activity analysis of compound 1 demonstrated improved antiaging (antioxidant, thermotolerance) properties against Caenorhabditis elegans, in comparison to a similar, commercially available molecule i.e., 1,5-pentanediol (2). The effective (lower) concentration of 1 significantly showed (28.6%) higher survival percentage of the worms under thermal stress (37 ºC) compared to its higher concentration (25.3%), while similar trends were followed in oxidative stress where (22.2%) higher survival percentage was recorded in comparison to untreated control. The compound 1, however, lacked potential antimicrobial activity, indicating the plausible ramification of the position of OH group in such bioactive molecules. In silico evaluation of these molecules against common as well as unique targets corroborated better antiaging potential of 1 in comparison to that of 2. The results for the first time indicated that the utilization of the endophytic fungi of C. amada could, thus, be a possible source for obtaining non-plant-based bioactive compounds having broader therapeutic applications pertaining to age-related progressions.

Antimicrobial polyacetylenes from Panax ginseng hairy root culture.

Two new polyacetylenes, 1-hydroxydihydropanaxacol (3) and 17-hydroxypanaxacol (4), were isolated from Panax ginseng hairy root culture, along with dihydropanaxacol (1), panaxacol (2) and ginsenoyne D (5). Highly hydroxylated compounds 1-4 were isolated from the medium and compound 5, which was a biosynthetic precursor of compound 1, was isolated from the roots. Compounds 1-4 showed antimicrobial activity against Staphylococcus aureus, Bacillus subtilis, Cryptococcus neoformans and Aspergillus fumigatus. It is suggested that P. ginseng plants release antimicrobial polyacetylenes into the surrounding soil from the roots as defense compounds.

Two new polyols and a new phenylpropanoid glycoside from the basidiomycete Lactarius deliciosus.

Two new polyols, 3-hydroxymethyl-2-methylenepentane-1,4-diol and 1-methylcyclohexane-1,2,4-triol, and a new phenylpropanoid glycoside, eugenyl 4″-O-acetyl-ß-rutinoside, together with seven known steroids (5-11) were isolated from the fruiting bodies of the basidiomycete Lactarius deliciosus. The structures of these compounds were elucidated by the analysis of spectroscopic data.

Male-produced aggregation pheromones of the cerambycid beetles Xylotrechus colonus and Sarosesthes fulminans.

Adults of both sexes of the cerambycid beetles Xylotrechus colonus (F.) and Sarosesthes fulminans (F.) were attracted to odors produced by male conspecifics in olfactometer bioassays. Analyses of headspace volatiles from adults revealed that male X. colonus produced a blend of (R)- and (S)-3-hydroxyhexan-2-one and (2 S,3 S)- and (2R,3R)-2,3-hexanediol, whereas male S. fulminans produced (R)-3-hydroxyhexan-2-one and (2 S,3R)-2,3-hexanediol. All of these compounds were absent in the headspace of females. Two field bioassays were conducted to confirm the biological activity of the synthesized pheromones: (1) enantiomerically enriched pheromone components were tested singly and in species-specific blends and (2) four-component mixture of racemic 3-hydroxyhexan-2-one plus racemic 2-hydroxyhexan-3-one and the four-component blend of the stereoisomers of 2,3-hexanediols were tested separately and as a combined eight-component blend. In these experiments, adult male and female X. colonus were captured in greatest numbers in traps baited with the reconstructed blend of components produced by males, although significant numbers were also captured in traps baited with (R)-3-hydroxyhexan-2-one alone or in blends with other compounds. Too few adult S. fulminans were captured for a statistical comparison among treatments, but all were caught in traps baited with lures containing (R)-3-hydroxyhexan-2-one. In addition to these two species, adults of two other species of cerambycid beetles, for which pheromones had previously been identified, were caught: Neoclytus a. acuminatus (F.) and its congener Neoclytus m. mucronatus (F.). Cross-attraction of beetles to pheromone blends of other species, and to individual pheromone components that are shared by two or more sympatric species, may facilitate location of larval hosts by species that compete for the same host species.

Identification of new molecules extracted from Quercus suber L. cork.

Various methods of suberin extraction have been used in order to identify monomers of this complex polymer. Pre-extraction of waxes has allowed us to identify for the first time 3-friedelanol as a terpen from cork. Moreover, the wax chemical composition found here varied from previous results since cerin was not identified while friedelin and betulin were. Three fractions were obtained: a polymeric, a monomeric and a low molecular weight fraction, the last of which has never before been described. 2,6-heptanediol was found to be the main compound of this fraction. Furthermore, depolymerisation at room temperature gives the same yields as those obtained at reflux, defining an easier and cheaper methodology.

Degradability and sediment sorption of an alcohol polyglycol ether surfactant putatively useful for the control of red swamp crayfish in rice fields.

This work reports studies of the degradation rates of a fatty alcohol polyglycol ether non-ionic surfactant, Genapol OXD-080, putatively useful for the control of red swamp crayfish (Procambarus clarkii Girard) in rice fields under laboratory and field conditions. The influence of temperature, sediment site specificity and sorption were taken into account. The degradation kinetics of the surfactant depends on the experimental conditions: type of inocula and temperature. The distribution of this chemical in aquatic systems was also examined. Genapol OXD-080 was removed into the sediments readily after application, and sorption was considered the major path of removal from the water phase. Data suggest that further studies are required regarding the effects of Genapol OXD-080 in aquatic organisms resident in rice fields, in parallel with the development of technologies related with the use of surfactants to control P. clarkii populations.

Infrared spectral analysis of extractables from poly(etherurethane urea) (PEUU) elastomers.

Poly(etherurethane urea) (PEUU) elastomers when employed as biomedical devices may be susceptible to extraction upon implantation. Four PEUU elastomers containing a single PEUU formulation, but varying in terms of their additives, were subjected to an in vitro extraction procedure. The additives in the PEUUs were Methacrol 2138 F at 5 wt% and Santowhite powder at 1 wt% levels. Only 1-2 wt% of the PEUUs was extractable with methanol. Fourier transform infrared spectroscopy (FT-IR) furnished qualitative and quantitative information on the extractables. The extractables consisted of a PEUU component that on the average was richer in soft segment than the bulk PEUU, and the two additives, Methacrol 2138 F and Santowhite powder.

A simplified methodology for quantitation of butadiene metabolites. Application to the study of 1,3-butadiene metabolism by rat liver microsomes.

A rapid, simple extraction and GC assay procedure is described that allows quantitation of micromolar concentrations of butadiene bisoxide and 3-butene-1,2-diol in microsomal suspensions exposed to butadiene. Butane-1,4-diol is used as the internal standard. The recovery of these compounds from aqueous media was almost quantitative, and calibrations for each compound were linear from 10(-6) to 10(-3) M. In this system butadiene monoxide and crotonaldehyde can be rapidly quantitated at micromolar concentration by headspace sampling, using methanol or n-butanol as the internal standard. In addition, the synthesis and chemical characterization of diastereomeric 3,4-epoxybutane-1,2-diols are described. It is demonstrated that the epoxy diol, although not extractable from aqueous solutions by ethyl acetate, can be recovered upon evaporation of aqueous media, followed by ethyl acetate extraction. Direct GC quantitation of the epoxy diol was linear from 10(-5) to 10(-3) M. This procedure facilitated the examination of butadiene metabolism by rat liver microsomes. Exposure of microsomes to butadiene resulted in the NADPH-dependent formation of butadiene monoxide, crotonaldehyde, 3-butene-1,2-diol, and one diastereomer of 3,4-epoxybutane-1,2-diol.

Concurrent separation of catecholamines, dihydroxyphenylglycol, vasoactive intestinal peptide, and neuropeptide Y in superfusate and tissue extract.

A method is described for separation and quantification of 3,4-dihydroxyphenylglycol (DO-PEG), norepinephrine (NE), dopamine (DA), vasoactive intestinal peptide (VIP), and neuropeptide Y (NPY) from single samples of tissue homogenate and from superfusate from in vitro dog blood vessel preparations using cartridges containing 0.4 g of octadecylsilane (Sep-Pak C-18). Samples were passed through the cartridge at pH 7.4. A step-gradient system was used to first selectively desorb the catechols (DOPEG, NE, DA) with a moderately polar eluent; subsequently VIP and NPY were eluted with 2.5 ml of a mixture of 1% trifluoroacetic acid, 80% acetonitrile. Five Sep-Pak catechol eluents were tested. Catechols were quantified by HPLC with electrochemical detection and peptides by radioimmunoassay. An HPLC solvent system is described which is particularly useful for chromatography of the more hydrophilic catechols DOPEG, 3,4-dihydroxymandelic acid, and 3,4-dihydroxyphenylalanine concurrently with catecholamines. For superfusion studies, sample cleanup time was reduced to about 4 min per sample by attachment of the cartridges directly to the bottom of the superfusion chamber. Superfusate was subsequently pulled through the cartridges immediately after they were passed over the tissue. Batches of 12 high-speed tissue supernates were processed through the method in about 30 min. The method was used to analyze DOPEG, NE, DA, VIP, and NPY in various rat and dog tissues. The values obtained were similar to values obtained previously by other methods. Because the catechols and peptides are separated from a single sample, the method has several advantages over those described previously; e.g., it is rapid, simple, and more sensitive.

A gas-liquid chromatographic method for quantitation of 1,3-butylene glycol in whole blood or plasma and the separation of the short chain glycols.

A microanalytical method with direct on-column specimen injection for determination of 1,3-butylene glycol (1,3-butanediol) in whole blood or plasma using gas-liquid chromatography with flame ionization is described. Whole blood or serum (minimum of 10 microL) was mixed with an equal volume of internal standard (1,2-propanediol, 50 mg/dL) and a 2-microL aliquot was injected onto the column without prior derivatization or extraction. The other short chain (C2 to C4) alkyldiols were separated by this method and did not interfere with the quantitation of 1,3-butylene glycol. The method was linear (y = 0.0206x + [-0.0073], r = 0.9990) over the range of 25 to 100 mg/dL and the coefficient of variation varied between 0.74 and 6.03%. Minimum detectable concentration of 1,3-butylene glycol was 5.0 mg/dL. The method described is suitable for the rapid detection of potentially toxic blood or plasma levels of 1,3-butylene glycol, as well as for the detection of other short chain glycols.

Mouse liver microsomal cholesterol epoxide hydrolase: a specific inhibition of its activity by 5,6 alpha-Imino-5 alpha-cholestan-3 alpha-OL.

A comparative study on mouse liver epoxide hydrolase activities has been done by using enzyme inhibitors in order to obtain evidence for the specificity of microsomal cholesterol epoxide hydrolase. 5,6 alpha-Imino-5 alpha-cholestan-3 beta-ol (IC) strongly inhibited the microsomal hydrolysis of cholesterol alpha-epoxide and the other delta 5-steroid alpha-epoxides (0.1 mM each) at concentrations less than 1 microM but affected neither microsomal nor cytosolic hydrolysis of any other epoxides of endogenous and exogenous compounds (0.1 mM each). On the other hand, 3,3,3-trichloropropene 1,2-oxide (TCPO) did not inhibited the microsomal hydrolysis of delta 5-steroid alpha-epoxides but strongly inhibited both microsomal and cytosolic hydrolysis of the other epoxides used. The only exceptions for the epoxy substrates that were not affected by both inhibitors were 5 alpha-cholest-2-ene alpha- and beta-epoxides. The inhibition by IC of the microsomal cholesterol alpha-epoxide hydrolysis was competitive, but no significant inhibition of the enzyme activity was observed by the typical microsomal xenobiotic substrates, hexadecene oxide and benzo[a]pyrene 4,5-oxide. These results strongly suggest that the microsomal enzyme hydrolyzing cholesterol alpha-epoxide differs from the microsomal hydrolase for epoxides of various xenobiotic olefins and arenes.

Bacterial conversion of phenylalanine and aromatic carboxylic acids into dihydrodiols.

Strain E of chloridazon-degrading bacteria, when grown on L-phenylalanine accumulates cis-2,3-dihydro-2,3-dihydroxyphenylalanine. In experiments with resting cells and during growth the bacterium converts the aromatic carboxylic acids phenylacetate, phenylpropionate, phenylbutyrate and phenyl-lactate into the corresponding cis-2,3-dihydrodiol compounds. The amino acids L-phenylalanine, N-acetyl-L-phenylalanine and t-butyloxycarbonyl-L-phenylalanine were also transformed into dihydrodiols. All seven dihydrodiols, thus obtained, were characterized both by conventional analytical techniques and by the ability to serve as substrates for a cis-dihydrodiol dehydrogenase.

Lipase hydrolysis of mammalian long-chain 1,2-alkanediol diesters. Nonrandom distribution of fatty acids.

Long-chain 1,2-alkanediol diesters were isolated from the total surface lipids of golden Syrian hamsters and Swiss albino mice. Hydrolysis of the diol diester waxes with exocellular lipase from Rhizopus arrhizus delemar or with purified porcine pancreatic lipase produced free fatty acids and 2-acyl diols in about 60--80% yield. Nonrandom distribution of the constituent fatty acids at positions 1 and 2 of the alkanediols was observed. In the diester waxes from the hamster, both straight-chain and branched-chain fatty acids of 14 to 20 carbon atoms predominated at position 1 and those of 22 to 26 carbon atoms at position 2. In contrast, the diester waxes of the mouse contained mainly fatty acids of less than 19 carbon atoms, both saturated and monounsaturated, at position 2 and those of greater chain length (20 to 24 carbon atoms) at position 1. The results of the lipase hydrolysis were confirmed by degradation of the diester waxes with Grignard reagent.

Thin-layer chromatographic separation of the 3-O-methylated metabolites of noradrenaline.

The use of sodium borate impregnated thin-layer silica gel plates for the separation of noradrenaline and its 3-O-methylated metabolites is described. Its application to studies of the metabolism of tritiated I-noradrenaline by isolated tissues is illustrated for the rabbit uterus.