Olefinic and acetylenic butenolides from Peucedanum alsaticum

The lipophilic root extract of Peucedanum alsaticum afforded four highly unstable butenolides which could be separated by HPLC. Whereas the stereochemistry of three olefinic derivatives, previously isolated only as their methyl ethers, were now established by ¹H NMR, the structure of a new acetylenic lactone was additionally confirmed by ¹³C NMR. The compounds are probably formed by condensation of unsaturated C₁₈-acids with pyruvate.     

Content of fatty acids ofChlorella kessleri in a deep tank fermentation

Chlorella kessleri cultivated in a deep tank contained 4.8% of non-polar lipid; 51% of this fraction represents saturated fatty acids, 7% unsaturated fatty acids. Our investigation of the fatty acids profile demonstrated even- and odd-numbered saturated and unsaturated fatty acids ranging from C₁₂ to C₂₀. Unlike in otherChlorella species, stearic acid was the dominant fatty acid found. Also shown was an elevated C_16:0 fatty acid content and a reduced level of unsaturated fatty acids.     

Synthesis of some acetylenic acids

All of the mono-ynoic acids (excluding both terminal and conjugated acetylenic acids) of chain length C₁₀ to C₁₄ have been prepared. None of these acids show any useful inhibition of the enzyme PG-Synthetase which is used in the synthesis of prostaglandins from arachidonic acid.     

Lipids of Fontinalis antipyretica

Fatty acid compositions of the lipids of Fontinalis antipyretica shows the presence of octadeca-9, 12-dien-6-ynoic, octadeca-9,12,15-trien- 6-ynoic and eicosa-11,14-dien-8-ynoic acids as the major components of the triglycerides. These acetylenic acids were not present in the galactosyl diglycerides. The moss lipids also contained relatively large amounts of two novel acetylenic methyl ketones with an unsaturation pattern analogous to that of the C₁₈ acetylenic acids.     

Echinacea alkamide disposition and pharmacokinetics in humans after tablet ingestion

Echinacea is a widely used herbal remedy for the treatment of colds and other infections. However, almost nothing is known about the disposition and pharmacokinetics of any of its components, particularly the alkamides and caffeic acid conjugates which are thought to be the active phytochemicals. In this investigation, we have examined serial plasma samples from 9 healthy volunteers who ingested echinacea tablets manufactured from ethanolic liquid extracts of Echinacea angustifolia and Echinacea purpurea immediately after a standard high fat breakfast. Caffeic acid conjugates could not be identified in any plasma sample at any time after tablet ingestion. Alkamides were rapidly absorbed and were measurable in plasma 20 min after tablet ingestion and remained detectable for up to 12 h. Concentration-time curves for 2,4-diene and 2-ene alkamides were determined. The maximal concentrations for the sum of alkamides in human plasma were reached within 2.3 h post ingestion and averaged 336+/-131 ng eq/mL plasma. No obvious differences were observed in the pharmacokinetics of individual or total alkamides in 2 additional fasted subjects who took the same dose of the echinacea preparation. This single dose study provides evidence that alkamides are orally available and that their pharmacokinetics are in agreement with the one dose three times daily regimen already recommended for echinacea.     

Bioactivity of alkamides isolated from Echinacea purpurea (L.) Moench.

Alkamides from the roots of Echinacea purpurea (L.) Moench were examined for anti-inflammatory activity in an in vitro model system. Cyclooxygenase-I (COX-I) and cyclooxygenase-II (COX-II) inhibitory activities were assessed at pH 7 for alkamides isolated from E. purpurea roots to compare inhibitory activities between the two cyclooxygenase isozymes. At 100 microg/ml, several E. purpurea alkamides inhibited COX-I and COX-II enzymes in the range of 36-60% and 15-46%, respectively, as compared to controls. Mosquitocidal activity was assessed at 100 and 10 microg/ml, with 100% mortality against Aedes aegyptii L. larvae noted for several E. purpurea alkamides at 100 microg/ml.     

Fatty Acids of Myxococcus xanthus

Fatty acids were extracted from saponified vegetative cells and myxospores of Myxococcus xanthus and examined as the methyl esters by gas-liquid chromatography. The acids consisted mainly of C₁₄ to C₁₇ species. Branched acids predominated, and iso-pentadecanoic acid constituted half or more of the mixture. The other leading component (11–28%) was found to be 11-n-hexadecenoic acid. Among the unsaturated acids were two diunsaturated ones, an n-hexadecadienoic acid and an iso-heptadecadienoic acid. No significant differences between the fatty acid compositions of the vegetative cells and myxospores could be detected. The fatty acid composition of M. xanthus was found to be markedly similar to that of Stigmatella aurantiaca. It is suggested that a fatty acid pattern consisting of a large proportion of iso-branched C₁₅ and C₁₇ acids and a substantial amount of an n-16:1 acid is characteristic of myxobacteria.     

Effects of Growth Pressure and Temperature on Fatty Acid Composition of a Barotolerant Deep-Sea Bacterium

The effects of pressure and temperature on the fatty acid composition in a barotolerant deep-sea bacterium that had branched-chain fatty acids were examined. The major fatty acids of the strain at atmospheric pressure were iso-C_15:0, C_16:1, iso-C_17:0, and iso-C_17:1. As the growth pressure increased, the proportion of unsaturated fatty acid increased because of an increase in the proportion of iso-C_17:1. On the other hand, as the growth temperature decreased, the proportion of unsaturated fatty acid increased because of the increase in the proportion of C_16:1 and C_18:1.     

Artepillin C,a Major Ingredient of Brazilian Propolis,Induces a Pungent Taste by Activating TRPA1 Channels

Brazilian green propolis is a popular health supplement because of its various biological properties. The ethanol extract of Brazilian green propolis (EEBP) is characteristic for its herb-like smell and unique pungent taste. However, the ingredients responsible for its pungency have not yet been identified. This study provides the first evidence that artepillin C is the main pungent ingredient in EEBP and that it potently activates human transient receptor potential ankyrin 1 (TRPA1) channels. EEBP was fractionated using column chromatography with a step gradient elution of an ethanol-water solution, and the fractions having the pungent taste were determined by sensory tests. HPLC analysis revealed that the pungent fraction was composed primarily of artepillin C, a prenylated derivative of cinnamic acid. Artepillin C was also identified as the pungent compound of EEBP by organoleptic examiners. Furthermore, the effects of artepillin C and other cinnamic acids found in EEBP on TRPA1 channels were examined by calcium imaging and plate reader-based assays in human TRPA1-expressing cells to investigate the molecular mechanisms underlying their pungent tastes. Artepillin C and baccharin activated the TRPA1 channel strongly, whereas drupanin caused a slight activation and p-coumaric acid showed no activation. Because the EC₅₀ values of artepillin C, baccharin, and allyl isothiocyanate were 1.8 µM, 15.5 µM, and 6.2 µM, respectively, artepillin C was more potent than the typical TRPA1 agonist allyl isothiocyanate. These findings strongly indicate that artepillin C is the main pungent ingredient in EEBP and stimulates a pungent taste by activating TRPA1 channels.     

Biosynthetically simple C18-alkamides from Achillea species

The petrol-diethyl ether extracts of the underground parts of Achillea lycaonica and A. chamaemelifolia afforded, in addition to known derivatives, 10 new alkamides. Their structures and stereochemistries were elucidated on the basis of spectroscopic evidence. Besides a new C₁₅-piperidide, the derivatives fall into a series of structurally closely related C₁₈-piperidides and -isobutylamides. A comparison of the different patterns of unsaturation allows some general conclusions about the biosynthetic pathways of the acid moieties.     

Structures of Monohydroperoxides Produced from Chlorophyll Sensitized Photooxidation of Methyl Linoleate

The effects on growth and mortality of larvae of pink bollworm (Pectinophora gossypiella, Saunders), bollworm (Heliothis zea, Boddie) and tobacco budworm (Heliothis virescens, F.) of adding selected C₁₀–C₁₂ fatty acid methyl esters to a standard diet were determined. The antibiotic activity of straight chain saturated esters was compared to the activity of esters with an olefinic bond either at C-2 or terminally or with a terminal acetylenic or cyclopropyl group. The ester with the greatest activity was the naturally occurring compound methyl (Z,Z)-deca-2,8-diene-4,6-diynoate (matricaria ester) which was lethal to all pink bollworm larvae at 0.01% in the diet and lethal to all bollworm and tobacco budworm larvae at 0.05%.     

Cis-unsaturated fatty acids modulate the function of the cell-surface cAMP receptor of Dictyostelium discoideum

The binding of cAMP to the chemotactic cAMP receptor in intact Dictyostelium discoideum cells and isolated membranes is strongly inhibited by unsaturated fatty acids. In isolated membranes, cis-unsaturated fatty acids decreased the number of accessible cAMP binding sites, without significantly altering their affinity. Most potent were C₁₈ and C₂₀ cis-poly unsaturated fatty acids, like arachidonic acid, linoleic acid and linolenic acid. Trans-unsaturated fatty acid was less potent than its cis isomer, while saturated fatty acids did not affect the binding of cAMP to receptors at all. Oxidation reactions were not important for the effect of unsaturated fatty acids. When membranes were preincubated with millimolar concentrations of Ca²⁺, the effect of unsaturated fatty acids was strongly diminished. Mg²⁺ was ineffective. Ca²⁺, if presented after the incubation of membranes with unsaturated fatty acids, did not reverse the inhibitory effect. The specificity of the fatty acid effect, and the interference with Ca²⁺, but not Mg²⁺, suggest that the properties of the cAMP receptor are changed as a result of alterations in the lipid bilayer structure of the membrane.     

Production of volatiles by degradation of lipids during manufacture of black tea

During manufacture of black tea, lipids are degraded to volatile constituents. Cis-3-hexenal was present in appreciable amounts in the various parts of fresh shoots and decreased in the second leaves during manufacture. There was a simultaneous increase in trans-2-hexenal. Linalol and methyl salicylate also increased appreciably during rolling and fermentation. Most of the volatiles were lost during the firing process. The above trend was borne out by the ‘potential’ of the leaves for the production of volatiles as indicated by the increased amounts of volatiles produced by homogenizing the tissue in water against controls homogenized in 0.1 N acid. The C₆-aldehydes present in the headspace of withered shoots increased significantly following mechanical damage. The major fatty acids of the lipids in the various parts of the shoots were linolenic, linoleic, palmitic, oleic and stearic acids. The ratio of linoleic to linolenic acid in the stems was much higher than that of the leaves or buds and this was reflected in its higher 'potential for formation of hexanal. During withering and rolling of the second leaves, the unsaturated fatty acids showed substantial losses compared with the saturated acids. It is suggested that the enzymic breakdown of membrane lipids initiate the formation of volatile carbonyl compounds which are partly responsible for the flavour of black tea.     

Quantitative effects of unsaturated fatty acids in microbial mutants. VII. Influence of the acetylenic bond location on the effectiveness of acyl chains.

The ability of a series of 18 carbon acetylenic fatty acids to fulfill the unsaturated fatty acid requirements of Escherichia coli and Saccharomyces cerevisiae was investigated. Despite their high melting points (greater than 40 degrees C), several isomers of the acetylenic fatty acids were as efficient or more efficient in supporting growth than the analogous fatty acid having a cis-double bond. The efficiencies of the different positional isomers in supporting cell proliferation varied from essentially 0 cells per fmol for the 2-5 and 13-17 isomers to high values when the acetylenic bond was near the center of the chain: e.g. 45 E. coli and 5.5 S. cerevisiae cells/fmol for the 10 isomer. A striking ineffectiveness of the 9 isomer was observed with E. coli. The 7, 8 and 10 isomers were at least 10-fold more efficient than any of the other positional isomers in supporting the growth of E. coli. In contrast, the 9 isomer was among the most effective acetylenic fatty acids tested with the yeast mutant. Chromatographic analysis of the extracted lipids indicated that each of the acetylenic isomers tested (except delta2 and delta3) could be esterified by the prokaryotic and eukaryotic microorganisms. The content of unsaturated plus cyclopropane acids observed when growth ceased in E. coli cultures supplemented with growth-limiting concentrations of the acetylenic fatty acids ranged from approx. 15 mol% for the 8 isomer to approx. 35 mol% for the 14 and 17 isomers. The 8-11 isomers were observed to be esterified predominantly at the two position in phosphatidylethanolamine of E. coli and in phosphatidylcholine of S. cerevisiae.     

Differences in lipid composition between free-living and initially adhered cells of a Gram-negative bacterium

The phospholipid fatty acid composition and poly--hydroxybutyrate (PHB) content of initially adhered and free-living cells of a Pseudomonas sp. isolated from the rape plant Brassica napus were examined with gas chromatography (GC). Five different adhesion experiments were made including variations in surface charge (hydrophilic and lipophilic), temperature, media composition and time of adhesion. Lipids and poly--hydroxybutyrate (PHB) were extracted with a chloroform-methanol-water mixture, hydrolyzed and esterified with pentafluorobenzyl bromide. Analysis was performed with capillary gas chromatography and flame ionization detection. A pronounced difference in both the ratio saturated/unsaturated fatty acids and in PHB content between free-living and adhered bacteria were found. The free-living bacteria has a significantly smaller ratio of saturated/unsaturated C₁₆ and C₁₈ fatty acids and also a smaller ratio of total C₁₈/total C₁₆ fatty acids. Bacteria adhered to the lipophilic surface had a higher ratio of saturated to unsaturated C₁₆ fatty acids than at the hydrophilic surface. There were no major differences between the treatments regarding the amount of bacteria adhered to the surface or their lipid composition.     

FATTY ACID ABNORMALITY IN ADRENOLEUKODYSTROPHY

—Recent clinical and morphological evidence established that adrenoleukodystrophy is a distinct X-linked genetic disorder. Fatty acid compositions of lipids in the brain, adrenal and serum from seven patients were examined. Cholesterol esters of both brain and adrenal contained substantial proportions of fatty acids longer than C₂₂ (11.8–41.9% of total in the brain and 13.4-34.8% of total in the adrenal), while cholesterol esters from normal and pathological control specimens contained very little. These very long chain fatty acids were generally saturated in brain cholesterol esters but significant amounts of unsaturated long chain fatty acids were also present in adrenal cholesterol esters. The long chain fatty acids showed bell-shaped distribution with C₂₅ or C₂₆ at the peak. Ganglio-sides from patients’white matter also showed increased proportions of very long-chain fatty acids, up to 50% of the total. Qualitatively similar but much milder fatty acid abnormalities were also found in galactosylceramide of the brain. On the other hand, fatty acids and fatty aldehydes of brain glycerophospholipids, adrenal free fatty acids, triglycerides and glycerophospholipids were not abnormal. Furthermore, serum cholesterol esters from two patients did not show the long-chain fatty acid abnormality found in brain and adrenal cholesterol esters. Sequential extractions with acetone and hexane established that the characteristic birefringent material in the brain and adrenal is indeed cholesterol esters with very long chain fatty acids. This type of fatty acid abnormality has not been described in other pathological conditions and may well represent the unique biochemical abnormality that is directly related to the fundamental genetic defect underlying adrenoleukodystrophy.     

Fatty acid composition of two Tunisian pine seed oils

Oils were extracted from fully ripen Pinus pinea L. and Pinus halepensis Mill seeds and fatty acid composition has been established by capillary gas chromatography. Seeds are rich in lipids, 34.63-48.12% on a dry weight basis. Qualitatively, fatty acid composition of both species is identical. For P. halepensis linoleic acid is the major fatty acid (56.06% of total fatty acids) followed by oleic (24.03%) and palmitic (5.23%) acids. For P. pinea, the same fatty acids are found with the proportions 47.28%, 36.56%, and 6.67%, respectively. Extracted fatty acids from both species are mainly unsaturated, respectively, 89.87% and 88.01%. Pinus halepensis cis-5 olefinic acids are more abundant (7.84% compared to 2.24%). Results will be important as a good indication of the potential nutraceutical value of Pinus seeds as new sources of fruit oils rich in polyunsaturated fatty acids and cis-5 olefinic acids.     

Fatty acid composition of lipids from differentiated tissues and cell cultures of Euonymus europaeus

The fatty acid patterns of Euonymus europaeus callus cultures and cell suspension cultures were analysed at the beginning of stationary growth phase and compared with those from the respective differentiated tissues. The lipid and fatty acid patterns in cell cultures differed remarkably from those in the tissues of the mother plant. No glycerol triacetate was detected in the callus cultures derived from differentiated tissues whereas in seeds this lipid compound amounts to 29%. In addition to fatty acids normally occurring in differentiated tissues, lipids in cultured cells also contained short-chain (C₁₂–C₁₄) as well as very long-chain fatty acids (C₂₀–C₂₄). In tissue culture cells the major fatty acids were found to be saturated, whereas in the mother cells unsaturated fatty acids were predominant. Palmitic acid is the most abundant fatty acid in most of the cultures. Lauric, myristic and palmitic acid amount to 50% in lipids of cell suspension cultures.     

Isolation and partial characterization of esters of trehalose from Corynebacterium ovis (C. pseudotuberculosis)

A glycolipid fraction was isolated from Corynebacterium ovis (C. pseudotuberculosis). It had [α]_D²⁵ = + 63.2° (C = 0.5, CHCl₃) and m.p. 43–46°C; the sugar content was 26%, determined as trehalose. Alkaline hydrolysis of the isolated fraction found trehalose as the sole water-soluble component, while glucose was found only after acid hydrolysis of the aqueous phase. Saturated and unsaturated short-chain mycolic acids with carbon atoms ranging from C₃₀ to C₃₆ were the constituents of the fatty acid moiety. The glycolipid fraction of C. ovis is therefore assumed to be a mixture of trehalose esters in which the trehalose molecule is esterified by saturated and unsaturated short-chain (C₃₀–C₃₆) mycolic acids.