Lipids and proteins in the Rathke's gland secretions of the North American mud turtle (Kinosternon subrubrum)

Lipids and proteins in the Rathke's gland secretions of the North American mud turtle (Kinosternon subrubrum, Kinosternidae) were analyzed by gas chromatography-mass spectrometry (GC-MS) and SDS-polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Analysis by GC-MS indicates 2,3-dihydroxypropanal and C₃–C₂₄ free or esterified fatty acids. Analysis by SDS-PAGE indicates a major protein component with an approximate molecular mass of 60 kDa and minor components ranging from ca. 23 to 34 kDa. The major component of K. subrubrum glandular secretions exhibits a mobility that matches that of the Kemp's ridley sea turtle (Lepidochelys kempi, Cheloniidae), suggesting that these proteins are evolutionarily conserved.     

Discrimination of <Emphasis Type="Italic">Oribotritia</Emphasis> species by oil gland chemistry (Acari,Oribatida)

The chemical composition of secretions from opisthonotal (oil) glands in four species of the oribatid mite genus Oribotritia (Mixonomata, Euphthiracaroidea, Oribotritiidae) was compared by means of gas chromatography—mass spectrometry. The secretions of all, O. banksi (from North America) and three Austrian oribotritiids (O. berlesei, O. hermanni, O. storkani), are shown to be based on certain unusual compounds, the iridoid monoterpenes chrysomelidial and epi-chrysomelidial and the diterpene β-springene. These components probably represent general chemical characteristics of oribotriid oil glands. Their relative abundance in the secretions along with further components (mainly saturated and unsaturated C₁₃-, C₁₅-, C₁₇-hydrocarbons, and the tentatively identified octadecadienal) led to well-distinguishable, species-specific oil gland secretions profiles. In addition a reduced set of “Astigmata compounds” (sensu Sakata and Norton in Int J Acarol 27:281–291, 2001)—namely the two monoterpenes neral and geranial—could be detected in extracts of O. banksi nevertheless indicating the classification of euphthiracaroids within the (monophyletic) group of “Astigmata compounds-bearing”-Oribatida. These compounds are considered to be apomorphically reduced in all Austrian species. Our findings emphasize the potential of chemosystematics using oil gland secretion profiles in the discrimination of morphologically very similar, syntopically living or even cryptic oribatid species.     

New chemicals from the dufour gland of the formicine ant Lasius niger (Hymenoptera:Formicidae)

A series of esters of C₈-C₁₃ fatty acids formed with C₁₀-C₁₂ straight chain alcohols, representing 3.7% of the glandular contents, was found in the Dufour gland of workers of the formicine ant Lasius niger. Altogether, nearly 50 components, among them hydrocarbons, alcohols, acetates and propionates were identified in the glandular liquid by means of gas chromatography (GC) and mass spectrometry (MS), using a solid injection technique. The wax esters and the propionates—decyl, dodecyl, octadecyl and farnesyl—have not been previously reported from ant secretions.     

A comparison of triglycerides from aphids and their cornicle secretions

Direct mass spectrometry of extracts showed that body triglycerides from 30 species of aphids contained the same fatty acid radicals, C₆ (hexanoyl), C_6:2 (sorboyl), C₁₄ (myristoyl), and C₁₆ (palmitoyl) as did the cornicle secretions, but in many species the proportions of hexanoyl and/or palmitoyl triglycerides were greater in the body. When cornicle secretions were collected progressively so as to draw increasingly upon body fat reserves, their composition changed gradually towards that of the body extracts.All summer forms of Myzus persicae had similar body triglycerides, even when selected for resistance to organophosphorus insecticides, or bred for 3 months on an artificial diet. The composition of body triglycerides was also independent of colour in two aphid species in which pink and green forms were compared.Body extracts contain enough triglycerides for their composition to be determined in single aphids and the use of body extracts allows examination of aphids lacking cornicles and of specimens that do not give cornicle secretion because of low body turgor. Although, as in the case of cornicle secretions, the triglyceride composition of body extracts was not well correlated with taxonomic position, body extracts provide a second chemical characteristic that can be used to define a particular species.     

Characterization of mycolic acids from the pathogen Rhodococcus equi by tandem mass spectrometry with electrospray ionization

We describe a simple tandem mass spectrometric approach toward structural characterization of mycolic acids, the long-chain α-alkyl-β-hydroxy fatty acids unique to mycobacteria and related taxa. On collisionally activated dissociation in a linear ion trap or tandem quadrupole mass spectrometer, the [M−H]⁻ ions of mycolic acid generated by electrospray ionization undergo dissociation to eliminate the meroaldehyde residue, leading to formation of carboxylate anions containing α-alkyl chains. The structural information from these fragment ions affords structural assignment of the mycolic acids, including the lengths of the meromycolate chain and the α-branch. This study revealed that the mycolic acids isolated from pathogenic Rhodococcus equi 103 contained a series of homologous ions having C₃₀ to C₅₀ chain with 0–2 double bonds. The α-branch ranged from C₁₀ to C₁₈ with 0 to 1 double bond, in which 16:0 and 14:0 are the most prominent, whereas the meromycolate chain ranged from C₁₄ to C₃₄ with 0 to 2 double bonds. The major molecular species consisted of more than 3 isomers that differ by the lengths of the α-branch or meromycolate chain, and up to 10 isobaric isomers were identified for some minor ions. We also employed tandem quadrupole mass spectrometry with precursor ion and neutral loss scans for profiling mycolic acid with specific structure in mixtures. The tandem spectra obtained from precursor ion scans of m/z 255 (16:0-carboxylate anion) and m/z 227 (14:0-carboxylate anion) may provide a simple specific means for classification of Rhodococci species, whereas tandem spectra from neutral loss of meroaldehyde residue scans provided a simple approach to reveal the mycolic acid molecules with specific meromycolate chain in mixtures.     

Esters of trehalose from Corynebacterium diphtheriae: A modified purification procedure and studies on the structure of their constituent hydroxylated fatty acids

The purification procedure of 6,6′-diesters of trehalose from Corynebacterium diphtheriae was modified and the isolated substance was analysed by mass spectrometry as its permethylated derivative. The fatty acid moiety released from the glycolipid after alkaline hydrolysis was studied by mass spectral analysis of the O-methylated and O-acetylated methyl ester derivatives. By argentation thin-layer chromatography, three species of O-acetylated methyl esters were recognized, corresponding to saturated, mono-unsaturated and di-unsaturated α-branched-β-hydroxylated fatty acids. The double bond was located by ozonolysis of the O-acetylated methyl ester derivatives, by gas chromatography of the reaction product and mass spectrometry of the effluent from the gas chromatograph. The main components of each species of α-branched-β-hydroxylated fatty acids found in the gly colipid fraction of C. diphtheriae were 2-tetradecyl-3-hydroxyoctadecanoic acid (C₃₂H₆₄O₃, corynomycolic acid), 2-tetradecyl-3-hydroxy-11-octadecenoic acid (C₃₂H₆₂O₃, corynomycolenic acid), 2-tetradec-7′-enyl-3-hydroxy octadecanoic acid (C₃₂H₆₂O₃) and 2-tetradec-7′-enyl-3-hydroxy-11-octadecenoic acid (C₃₂H₆₀O₃, corynomycoldienic acid). The glycolipid fraction from C. diphtheriae is obviously a complex mixture of 6,6′-diesters of trehalose.     

Mono- and digalactosyldiacylglycerol composition of dinoflagellates. III. Four cold-adapted,peridinin-containing taxa and the presence of trigalactosyldiacylglycerol as an additional glycolipid

Despite their importance in marine and freshwater microalgal assemblages, cold-adapted dinoflagellates have been the subject of few comprehensive lipid studies, particularly with respect to those lipids that comprise plastid membranes. In an effort to understand the differences between warm- and cold-adapted dinoflagellate glycolipid composition, four peridinin-containing, cold-adapted dinoflagellates were surveyed for intact forms of monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), two common plastid lipids, using positive-ion electrospray ionization/mass spectrometry (ESI/MS) and electrospray ionization/mass spectrometry/mass spectrometry (ESI/MS/MS). It was determined that the dominant forms of MGDG and DGDG in these cold-adapted, peridinin-containing dinoflagellates possessed C₁₈ fatty acids and did not, with the exception of a 20:5/18:5 form of DGDG in a cold-adapted Gymnodinium sp. from the Baltic Sea, have C₂₀ fatty acids. This finding is in contrast to an earlier study of 35 peridinin-containing, warm-adapted dinoflagellates, which discovered a cluster dominated by C₁₈ fatty acids and a cluster dominated by both C₂₀ and C₁₈ fatty acids. The key difference in MGDG and DGDG production between the former group and the cold-adapted dinoflagellates examined in this study is that the cold-adapted species’ DGDG fatty acids were less saturated. Each cold-adapted dinoflagellate possessed both 18:5/18:5 and 18:5/18:4 DGDG, while most of the warm-adapted dinoflagellates contained only 18:5/18:4 DGDG. This survey also revealed the presence of a putative 18:1/14:0 trigalactosyldiacylglycerol (TGDG) as a dominant glycolipid in Gymnodinium sp. TGDG, previously unreported in dinoflagellates, was also discovered in Gymnodinium sp. in the forms of 18:1/16:0 and 18:1/18:1 TGDG, as minor lipids. Since the fatty acids associated with TGDG are not those found with dominant forms of MGDG or DGDG, TGDG may be produced by a different biosynthetic pathway.     

Changes in molecular species composition of nocardomycolic acids in nocardia rubra by the growth temperature

Nocardomycolic acids from Nocardia rubra were fully separated and characterized by a combination of argentation thin-layer chromatography and gas chromatography — mass spectrometry (GCMS). The occurrence of 20 or more different molecular species of mycolic acids was demonstrated. GCMS analysis of each subclass of mycolic acids after separation on AgNO₃ thin-layer chromatography revealed that in general the major species consisted of the even-carbon mycolic acids ranging from C₃₈ to C₅₂. However, the most abundant species differed by the subclasses; C₄₄ being in saturated, C₄₆ in monoenoic and C₄₆ in dienoic mycolic acids, respectively. All these acids were shown to possess C₁₂ or C₁₄ alkyl branch at 2 position, while double bonds were located in longer straight chain alkyl unit.By using this method, distinctive changes in mycolic acid composition by growth temperature were observed. The ratios of saturated, monoenoic to dienoic mycolic acids in a mixture of certain carbon numbered mycolic acids varied greatly, according to the shift of growth temperature. The mass fragmentographic analysis, monitoring M-15 ions derived from the loss of methyl group from the molecular ions showed the lower temperature (15°C) grown cells contained more unsaturated (especially dienoic) mycolic acids, while the higher temperature (40°C) grown cells contained more saturated mycolic acids in both extractable and cell-wall bound lipids. These changes in mycolic acid composition occurred shortly after shifting up the growth temperature from 20°C to 43°C at a logarithmic stage of the bacterial growth.     

Chemical composition of cornicle secretion of the brown citrus aphid Toxoptera citricida

Brown citrus aphid Toxoptera citricida Kirkadly is considered as an important pest of citrus because it vectors citrus tristeza closterovirus. Aphids secrete a fluid from their cornicles as a defensive mechanism against natural enemies. Earlier studies on cornicle secretions of aphids focus only on triglycerides and fatty acids. In the present study, three different methods are used to investigate the chemical composition of the cornicle fluid of T. citricida. Gas chromatography with flame ionization detection is used to detect and quantify the triglycerides after trimethylsilyl derivatization, and gas chromatography‐mass spectrometry (GC‐MS) is used to determine the fatty acid composition after derivatization with boron trifluoride–methanol. Other compounds are detected using GC‐MS after methoxyamine hydrochloride and N‐methyl‐N‐(trimethylsilyl)trifluoroacetamide derivatization. The major fatty acid in the cornicle secretion of T. citricida is palmitic acid. Oleic, stearic, myristic, myristoleic and sorbic acids are also detected, although in low amounts. Sorboyl, dipalmitoyl (C_6‐2, C₁₆, C₁₆) and disorboyl, stearoyl (C_6‐2, C_6‐2, C₁₈) are the main triglycerides detected in cornicle secretion. Trehalose is the most predominant sugar (558.2 mm ), followed by glucose (92.0 mm ) and inositol (48.8 mm ). Many amino acids, including proline, glycine, alanine and serine, are also detected. In addition, the cornicle secretion is rich in many organic acids, including malic, citric, succinic and lactic acid. Information obtained from the present study improves our understanding of the chemical composition of the cornicle secretion of the brown citrus aphid.     

Distribution of C22-, C24- and C26-α-Unit-Containing Mycolic Acid Homologues in Mycobacteria

There are three mycolic acid homologues with C₂₂-, C₂₄- and C₂₆-α-units in Mycobacterium. In order to reveal the composition and distribution of these homologues in each subclass and molecular species of mycolic acids and to compare them with the composition of constitutive non-polar fatty acids (free and bound forms), we have separated non-polar fatty acids and each subclass of mycolic acids from 21 mycobacterial species by thin-layer chromatography, and analyzed non-polar fatty acid methyl esters by gas chromatography (GC) and the cleavage products of methyl mycolate by pyrolysis GC. We further performed mass chromatographic analysis of trimethylsilyl (TMS) ether derivatives of mycolic acid methyl esters by monitoring [B-29]⁺ ions (loss of CHO from the α-branched-chain structure of mycolic acids) of m/z 426, 454 and 482 which are attributed to C₂₂-, C₂₄- and C₂₆-α-units of TMS ether derivatives of methyl mycolates, respectively, (Kaneda, K. et al, J. Clin. Microbiol. 24: 1060-1070, 1986). By pyrolysis GC, C_22:0, C_24:0 and C_26:0 fatty acid methyl esters generated by the C₂-C₃ cleavage of C₂₂-, C₂₄- and C₂₆-α-unit-containing mycolic acid methyl esters, respectively, were detected. Their proportion was almost the same among subclasses of mycolic acids in every Mycobacterium and also similar to the proportion of constitutive non-polar C_22:0, C_24:0 and C_26:0 fatty acids. By mass chromatography, the composition and distribution of C₂₂- and C₂₄-α-unit-containing homologues were revealed to be similar between α- and α'-mycolic acids in every Mycobacterium. We further analyzed in detail M. vaccae and demonstrated that the mass chromatogram of C₂₂-α-unit-containing homologue was analogous in shape to that of the C₂₄-α-unit-containing one, with the latter mass chromatogram being up-shifted from the former by two carbon numbers, in every subclass of α-, α'-, keto and dicarboxy mycolic acids. The present study suggests that the compositions of three homologues of both mycolic acids and constitutive non-polar fatty acids, which are characteristic to each mycobacterial species, may reflect the proportion of the amount of free C_22:0, C_24:0 and C_26:0 fatty acids synthesized in the cell. It is further demonstrated that intermolecular condensation of two fatty acids which become α- and β-units of mycolic acids will occur independently of the carbon chain length or kinds of polar moieties of fatty acids.     

Comparative studies on simple lipids ofSclerotium cepivorum,the causal agent of white rot of onion,and other fungi of the class Deuteromycetes

Summary The simple lipids ofSclerotium cepivorum, the causal agent of white rot of onion and nine other fungal species of the same class were investigated. The fatty acid composition of the simple lipids of these fungi were determined by GLC. The main fatty acids common to these fungal species were C₁₆ (saturated) and C₁₈ (unsaturated) acids. The sterol fraction was isolated by column chromatography and its components were detected by GLC and mass spectrometry. Ergosterol and γ-Ergostenol were found mostly in all fungal species under investigation. However, two fungal species namelyAlternaria alternata andScolecobasidium constrictum showed no Ergosterol.     

Lipids in femoral gland secretions of male lizards, Psammodromus hispanicus

Many lizards produce chemical secretions that may be used as pheromones in reproductive behavior, but only a few studies have identified chemical compounds in secretions. By using GC–MS, we found only 20 lipophilic compounds in femoral glands secretion of male lizards, Psammodromus hispanicus. Main compounds were six steroids (mainly cholesterol and campesterol) and seven n-C₉ to n-C₁₈ carboxylic acids (especially dodecanoic acid), and minor components were six alcohols between C₁₆ and C₂₉ and squalene. We compared these chemicals with those previously found in secretions of the closely related sister species Psammodromus algirus and other lizard species.     

The distribution of fatty acids including petroselinic and tariric acids in the fruit and seed oils of the Pittosporaceae, Araliaceae, Umbelliferae, Simarubaceae and Rutaceae

The fatty acid composition of the fruit oils or seed oils of Pittosporaceae (eight genera, 10 species), Araliaceae (two species), Simarubaceae (three species), and of one umbelliferous and one rutaceous species were determined by gas chromatography, argentation TLC and ozonolysis. In the Pittosporaceae, in which the major C₁₈ fatty acid of all species was either oleic acid (18:1, 9c) or linoleic acid (18:2, 9c, 12c), large amounts of C₂₀ and C₂₂ fatty acids seem to occur regularly. Petroselinic (18:1, 6c) and tariric (18:1, 6a) acids were absent. However, petroselinic acid was the major fatty acid in the Araliaceae and Umbelliferae. In these two families only small amounts of C₂₀ and C₂₂ acids were detected and tariric acid was absent. The Rutales contained relatively high amounts of trans-octadecenoic acids (18:1, 9t). Tariric acid was the major fatty acid in the two species of Picramnia (Simarubraceae), which also contained small amounts of petroselinic acid. The major fatty acids in Ailanthus glandulosa (Simarubaceae) and Phellodendron amurense (Rutaceae) were linoleic or linolenic acid (18:3, 9c, 12c, 15c); these species contained neither tariric nor petroselinic acid and the levels of C₂₀ and C₂₂ fatty acids were low. The appearance of schizogenous resin canals and polyacetylenes and the absence of iridoids and petroselinic acid allows the Pittosporaceae to be separated from the Rutales and Araliales and to be placed in an independent order, the Pittosporales. Arguments for a rather close relationship of the Pittosporales to the Araliales and Cornales (including the Escalloniaceae) are presented.     

Composition of Lipid-derived Polymers from Different Anatomical Regions of Several Plant Species

The composition of the aliphatics of the protective cuticular polymers from different anatomical regions from several plant species was determined by combined gas-liquid chromatography and mass spectrometry of the depolymerization products derived from the polymers. The polymer from the aerial parts of Vicia faba showed similar composition; dihydroxypalmitic acid was the major (>85%) component of the cutin covering leaves, petioles, flower petals and stem with smaller amounts of palmitic acid and ω-hydroxy palmitic acid. On the other hand, the chief components of the polymer from the tap root were ω-hydroxy C_16:0 and C_18:1 acids and/or the corresponding dicarboxylic acids. The positional isomer composition of the dihydroxy C₁₆ acids was shown to be dependent upon anatomical location, developmental stage, and light. Apple cutin from rapidly expanding organs (flower petal and stigma) was shown to contain predominately C₁₆ family acids whereas the C₁₈ family dominated in cutin of slower growing organs (leaf and fruit). The composition of the aliphatic components of cutin found in the seed coats of pea, corn, barley, and lettuce was found to be similar to that of the cuticular polymer of the leaves in each species.     

Identification by gas chromatography and mass spectrometry of lipids from the rat Harderian gland

Lipids from rat Harderian glands were extracted with ethyl acetate, hydrolysed with base and examined by gas chromatography (GC) and gas chromatography—mass spectrometry (GC—MS) as trimethylsilyl (TMS), [²H₉]TMS, methyl ester—TMS, picolinyl, nicotinate and nicotinylidene derivatives. The latter three derivatives were used to reveal the structures of the alkyl chains of fatty acids, alcohols and glycerol ethers, respectively. Forty-eight compounds were identified, representing about 97% of the total extracted lipids as measured by GC peak areas. The major constituents were fatty acids with chain lengths from 12 to 22 carbon atoms (mainly C₁₈ and C₂₀) and fatty alcohols (C₁₆ to C₂₆) derived from wax esters. Most of these acids and alcohols were unsaturated in the ω-7 position and were accompanied by smaller amounts of the saturated and ω-5 monounsaturated analogues. Glycerol ethers were also identified for the first time in this secretion; the ether chains contained from 14 to 19 carbon atoms (mainly 16) and were straight-chain saturated, unsaturated (ω-5 and ω-7) and branched (iso). The only sterol found was cholesterol amounting to 1.24% of the total extract.     

Enhancement of interferon-β production with sphingomyelin from fermented milk

A fermented milk, Kefir, contains an active substance which enhances IFN- secretion of a human osteosarcoma line MG-63 treated with a chemical inducer, poly I: poly C. The active substance in the fermented milk was identified to be sphingomyelin (SpM) by a combined use of a fast atom bombardment mass spectrometry (FAB-MS) and a fast atom bombardment tandem mass spectrometry (FAB-MS/MS). SpM from fermented milk (F-SpM) was a mixture of four molecular species of SpMs having C₂₁-, C₂₂-, C₂₃- and C₂₄-fatty acids. F-SpM enhanced the IFN secretion 14 times, SpMs from other sources also enhanced moderately (2–3 times). Sphingosine and lysosphingomyelin also enhanced the activity but ceramide and cerebroside did not.Abbreviations IFN- interferon- - SpM sphingomyelin - Lyso-SpM lysosphingomyelin - SpS sphingosine - FAB-MS fast atom bombardment mass spectrometry - FAB-MS/MS fast atom bombardment tandem mass spectrometry     

Species Specificity of Aldehyde and Fatty Acid Profiles of Four Family Group Representatives within the Insect Infraorder Pentatomomorpha (Hemiptera: Heteroptera)

Reactive α,β‐unsaturated aldehydes, including 4‐oxoalk‐2‐enals, are known to be present in volatile secretions of numerous heteropteran insect species. Because the aldehydes are likely to originate from metabolism of fatty acids (FAs), the present study aimed to examine and compare the aldehyde and FA profiles of four model heteropteran species. The model species consisted of adult family group representatives within the infraorder Pentatomomorpha (Hemiptera: Heteroptera): seed bug (Lygaeus equestris (Lygaeoidea)), dock leaf bug (Coreus marginatus (Coreoidea)), red firebug (Pyrrhocoris apterus (Pyrrhocoroidea)), and European stink bug (Graphosoma lineatum (Pentatomoidea)). Solid‐phase microextraction combined with two‐dimensional gas‐chromatography/time‐of‐flight mass spectrometry was used to establish the profiles of volatile secretions in stressed living insects. The FA profiles of acylglyceride and phospholipid fractions deposited in fat body and/or hemolymph were obtained by liquid chromatography/mass spectrometry and gas chromatography with flame ionization detection techniques. Our results based on multivariate statistical analyses of the data imply that volatile secretion blends as well as fat body and/or hemolymph lipid profiles are species specific but the differences in volatile blends between different species do not mirror the changes in corresponding fat body and/or hemolymph lipid profiles of stressed and non‐stressed individuals.     

The composition of suberin from the corks of Quercus suber L. and Betula pendula roth

The suberin constituents of Quercus suber and Betula pendula have been isolated after alkaline hydrolysis of the corks and over 80% by weight identified using thin-layer chromatography, preparative thin-layer chromatography, gas-liquid chromatography and combined gas chromatography — mass spectrometry. Long-chain aliphatic acids ranging from C₁₆–C₂₆ comprise about 90% of both suberin fractions; monobasic, α,ω-dibasic, ω-hydroxymonobasic, dihydroxymonobasic, dihydroxydibasic and trihydroxymonobasic acid classes are present. The principal suberin acids of Q. suber are 18-hydroxyoctadecenoic (12%), 22-hydroxydocosanoic (25%), 9,10-dihydroxyoctadecane-1,18-dioic (15%) and 9,10,18-trihydroxyoctadecanoic (8%), and those of B. pendula 9,10,18-trihydroxyoctadecanoic (43%) and 22-hydroxydocosanoic (16%).     

Quorum sensing in halophilic bacteria: detection of N-acyl-homoserine lactones in the exopolysaccharide-producing species of Halomonas

Some members of the moderately halophilic genus Halomonas, such as H. eurihalina, H. maura, H. ventosae and H. anticariensis, produce exopolysaccharides with applications in many industrial fields. We report here that these four species also produce autoinducer molecules that are involved in the cell-to-cell signaling process known as quorum sensing. By using the N-acyl homoserine lactone (AHL) indicator strains Agrobacterium tumefaciens NTL4 (pZRL4) and Chromobacterium violaceum CV026, we discovered that all the Halomonas strains examined synthesize detectable AHL signal molecules. The synthesis of these compounds was growth-phase dependent and maximal activity was reached during the late exponential to stationary phases. One of these AHLs seems to be synthesized only in the stationary phase. Some of the AHLs produced by H. anticariens FP35^T were identified by gas chromatography/mass spectrometry and electrospray ionization tandem mass spectrometry as N-butanoyl homoserine lactone (C₄-HL), N-hexanoyl homoserine lactone (C₆-HL), N-octanoyl homoserine lactone (C₈-HL) and N-dodecanoyl homoserine lactone (C₁₂-HL). This study suggests that quorum sensing may also play an important role in extreme environments.     

Identification of the endogenous apolar ecdysteroid conjugates present in newly-laid eggs of the house cricket (Acheta domesticus) as 22-long-chain fatty acyl esters of ecdysone

Newly-laid eggs of the house cricket Acheta domesticus contain significant amounts of apolar ecdysteroid conjugates, which can be hydrolysed by prolonged incubation with a mixture of Helix pomatia gut hydrolases. The ecdysteroid released on hydrolysis of the apolar conjugates has been purified and identified as ecdysone by co-chromatography on normal-phase and reversed-phase HPLC and by fast-atom bombardment mass spectrometry.Starting with only 22 g newly-laid eggs (containing 16 μg conjugated ecdysone), the ecdysone conjugates have been purified by open column chromatography and four successive HPLC purification steps to give essentially pure apolar conjugates with a yield of 57%. The conjugates are shown to be a mixture of ecdysone 22-fatty acyl esters by co-chromatography with authentic reference compounds and by fast-atom bombardment mass spectrometry. The fatty acyl composition of the conjugates is very similar to that produced by the ovaries of A. domesticus from [³H]ecdysone in vitro (Whiting and Dinan, Biochem. J.252, 95–103, 1988). The major fatty acyl esters are the 22-palmitate (C_16:0), 22-oleate (C_18:1) and 22-linoleate (C_18:2), with smaller amounts of the myristate (C_14:0), stearate (C_18:0) and arachidate (C_20:0) esters.This report constitutes the first identification from an insect source of endogenous ecdysteroid 22-fatty acyl esters, which have previously been identified in ticks and as metabolites of exogenous [³H]ecdysone in several arthropod species.