Developmental Changes in the Sterol Composition and the Glycerol Content of Cuticular and Internal Lipids of Three Species of Flies

The glycerol concentration and the composition of cuticular and internal sterols in three medically and forensically important fly species, viz., Musca domestica, Sarcophaga carnaria, and Calliphora vicina, were analyzed. The cuticular and internal lipid extracts were separated by HPLC‐LLSD, after which the sterol fraction was characterized by GC/MS in total ion current (TIC) mode. The cuticular lipids of M. domestica larvae contained seven sterols, while in pupae and females, six sterols were identified. Five sterols were found in the cuticular lipids of M. domestica males. The internal lipids of M. domestica larvae and pupae contained six and seven sterols, respectively, while those of male and female flies contained only five sterols. Sitosterol, cholesterol, and campesterol were the dominant sterols in M. domestica, while campestanol, stigmasterol, sitostanol, and fucosterol were identified in low concentrations or in traces. In contrast, cuticular and internal lipids of S. carnaria and C. vicina contained only cholesterol. Glycerol was identified in all stages of M. domestica, S. carnaria, and C. vicina. For all the three examined fly species, the present study clearly showed species‐specific developmental changes in the composition of cuticular and internal sterols as well as in the glycerol concentration.     

The effect of the environment on the permeability and composition of Citrus leaf cuticles

The constituents of the soluble cuticular lipids (SCL) of the leaf blades of Citrus aurantium L. were identified by gas chromatography-mass spectrometry and quantified. Major components were 1-alkanols (C₂₄ to C₄₀), n-alkyl esters (C₃₆ to C₅₆), n-alkanoic acids (C₂₈ to C₃₄), n-alkanes (C₂₂ to C₄₀) and triterpenones, while n-alkanals (C₂₉ to C₃₈), sterols, and alkyl benzenes (molecular weights 260, 274 and 288) made minor contributions. Leaf age and side significantly affected the quantitative composition of SCL. Increased day temperature during the development of leaves led to decreased amounts per unit area of n-alkanes, 1-alkanols, n-alkanoic acids and n-alkyl esters while increased night temperatures resulted in increased amounts of n-alkanes n-alkanoic acids and 1-alkanols. Relative humidity had no effect on the amounts or composition of SCL. The permeability of cuticular membranes to water (described in part I of this paper) and the composition of SCL were not related. A model for the molecular structure of the transport-limiting barrier of plant cuticles and for the transport of water across it is proposed.Abbreviations CM cuticular membrane - GC gas chromatogra-phy - MS mass spectroscopy - TLC thin-layer (planar) chromatography - SCL soluble cuticular lipids The authors are indebted to Dr. R. Winkler and H. Krause, Laboratorium für Strukturchemie des Fachbereichs Chemie, Biologie und Geowissenschaften, Technische Universität München, FRG, for performing the GC-MS analyses and their valuable help in the identification of SCL constituents. This work has been supported by the Deutsche Forschungsgemeinschaft and the Bayerische Staatsministerium für Wissenschaft und Kunst.     

The effect of the entomopathogenic fungus Conidiobolus coronatus on the composition of cuticular and internal lipids of Blatta orientalis females

The composition of cuticular and internal lipids in females of the cockroach Blatta orientalis L. exposed to the entomopathogenic fungus Conidiobolus coronatus is investigated. The compositions of the fatty acids, n‐alkanes, alcohol, sterols and methyl esters in the lipids are chemically characterized. Although contact with virulent colonies of the fungus does not induce insect mortality, significant changes in the lipid profiles, both cuticular and internal, are found. The cuticular extracts of a control group of B. orientalis females contain 24 compounds varying in carbon chain length from C6 to C22. The main cuticular fatty acids identified are: C16:1, C16:0, C18:1 and C18:0_. The cuticular lipids of B. orientalis females after exposure to C. coronatus contain only 14 free fatty acids from C8 to C20. The highest concentrations identified are C16:0, C18:2 and C18:1. Analysis by gas chromatography‐mass spectrometry identifies the presence of a homologous series of n‐alkanes containing from 25 to 31 carbon atoms. In the case of the insects after fungal exposure, the content of the n‐alkanes in the cuticular lipid is two‐fold higher compared with the controls. Of the cuticular lipids, 11 alcohols are found, ranging from C12:0 to C20:0. There is no presence of alcohols in the internal lipids of the control B. orientalis females and in all of the extracts from the B. orientalis females after fungal exposure. In the samples analyzed, the most common sterol is cholesterol. This is present in the cuticular lipids and the internal lipids of all of the insects sampled. The cuticular and internal lipids of females contain five fatty acid methyl esters, ranging in size from C15 to C19.     

Composition of soluble cuticular lipids and water permeability of cuticular membranes from Citrus leaves

Water permeability and composition of soluble cuticular lipids of isolated cuticular membranes from leaves of Citrus aurantium L. were investigated for 3 successive years. The average water permeability coefficient determined using 169 cuticular membranes was 1.09·10^–7 cm s^–1 with a standard deviation of 0.78·10^–7 cm s^–1. There were no significant differences in water permeability between years. Cuticular membranes are characterized by a great variability in water permeability both within and between years. Both water permeability of individual membranes and variability between membranes are shown to be determined by soluble cuticular lipids contained within the cuticular membranes. The soluble cuticular lipids of Citrus leaves are composed of fatty acids, primary alcohols, esters, and hydrocarbons. They occur in amounts of 9.84 g cm^–2, which represents approx. 3% of the total mass of isolated cuticular membranes. The specific weight of cuticular membranes (365.4 g cm^–1) and total amount of soluble cuticular lipids did not vary significantly between years. Significant differences were observed for the amounts and composition of the constituent classes of lipids. Six homologues comprise 86% of the fatty acids (C₁₆; C₁₈; C₁₉; C₂₁; C₂₄; C₂₆), 83% of the primary alcohols (C₂₄; C₂₆; C₂₈; C₃₀; C₃₂; C₃₄) and 88% of the esters (C₃₆; C₃₈; C₄₀; C₄₁; C₄₂; C₄₄). Eleven major homologues amount only to 62% of the total hydrocarbons (C₁₆; C₁₇; C₁₈; C₂₀; C₂₆; C₂₇; C₂₉; C₃₀; C₃₁; C₃₂; C₃₃). Variability in the composition of soluble cuticular lipids between years was much smaller than variability of water permeability and, therefore, no relation between composition of soluble cuticular lipids and water permeability could be found. It is suggested that this may be due to the fact that the lipid composition observed represents the averages of 20 to 30 membranes analyzed so that differences between individual membranes may have been leveled out.Abbreviations CM cuticular membranes - MX polymer matrix - P_d permeability coefficient for diffusion of water - SCL soluble cuticular lipids - MES morpholinoethane sulphonic acid     

Variations of the Composition of the Leaf Cuticular Wax among Chinese Populations of Plantago major

Plantago major L. grows in a very wide range of regions in China and exhibits great variations among populations. The analysis of the cuticular‐wax composition provides a potential approach to classify populations of P. major confronting different environmental conditions. Twelve populations of P. major and five populations of P. depressa Willd ., distributed over regions with average annual temperatures ranging from ?2.0 to 18.4°, were sampled, the variation of the composition of their cuticular waxes was analyzed, and their values of average chain length (ACL) and carbon preference index (CPI) were calculated. Great intra‐ and interspecies variations were observed for the total wax contents. The average annual temperature of the habitats was significantly correlated with the relative contents of the dominant n‐alkanes with an odd number of C‐atoms, but not with the wax contents. With an increasing average annual temperature, the relative contents of n‐alkanes C₂₉ and C₃₁ decreased, whereas those of C₃₃ and C₃₅ as well as the values of ACL_total and ACL_27–33 increased. Cluster analysis based on the pattern of the n‐alkane distribution allowed to clearly separate the populations of P. major according to the average annual temperature of their habitats, but not to separate the populations of the two species. Hence, the pattern of the n‐alkane distribution might be a good taxonomic marker for P. major at the intraspecies level, but not at the interspecies level. Nevertheless, a small difference between the populations of the two species was observed concerning the values of ACL_total and CPI_total, implying the potential use of these indices for the classification of the populations of the two species at the interspecies level.     

Leaf n‐Alkanes as Characters Differentiating Coastal and Continental Juniperus deltoides Populations from the Balkan Peninsula

The composition of the cuticular n‐alkanes isolated from the leaves of nine populations of Juniperus deltoides R.P.Adams from continental and coastal areas of the Balkan Peninsula was characterized by GC‐FID and GC/MS analyses. In the leaf waxes, 14 n‐alkane homologues with chain‐lengths ranging from C₂₂ to C₃₅ were identified. n‐Tritriacontane (C₃₃) was dominant in the waxes of all populations, but variations between the populations in the contents of all n‐alkanes were observed. Several statistical methods (ANOVA, principal component, discriminant, and cluster analyses) were used to investigate the diversity and variability of the cuticular‐leaf‐n‐alkane patterns of the nine J. deltoides populations. This is the first report on the n‐alkane composition for this species. The multivariate statistical analyses evidenced a high correlation of the leaf‐n‐alkane pattern with the geographical distribution of the investigated samples, differentiating the coastal from the continental populations of this taxon.     

Cuticle hydrolysis in four medically important fly species by enzymes of the entomopathogenic fungus Conidiobolus coronatus

Entomopathogenic fungi infect insects via penetration through the cuticle, which varies remarkably in chemical composition across species and life stages. Fungal infection involves the production of enzymes that hydrolyse cuticular proteins, chitin and lipids. Host specificity is associated with fungus–cuticle interactions related to substrate utilization and resistance to host‐specific inhibitors. The soil fungus Conidiobolus coronatus (Constantin) (Entomophthorales: Ancylistaceae) shows virulence against susceptible species. The larvae and pupae of Calliphora vicina (Robineau‐Desvoidy) (Diptera: Calliphoridae), Calliphora vomitoria (Linnaeus), Lucilia sericata (Meigen) (Diptera: Calliphoridae) and Musca domestica (Linnaeus) (Diptera: Muscidae) are resistant, but adults exposed to C. coronatus quickly perish. Fungus was cultivated for 3 weeks in a minimal medium. Cell‐free filtrate, for which activity of elastase, N‐acetylglucosaminidase, chitobiosidase and lipase was determined, was used for in vitro hydrolysis of the cuticle from larvae, puparia and adults. Amounts of amino acids, N‐glucosamine and fatty acids released were measured after 8 h of incubation. The effectiveness of fungal enzymes was correlated with concentrations of compounds detected in the cuticles of tested insects. Positive correlations suggest compounds used by the fungus as nutrients, whereas negative correlations may indicate compounds responsible for insect resistance. Adult deaths result from the ingestion of conidia or fungal excretions.     

Chemotaxonomic Implications of the n‐Alkane Composition and the Nonacosan‐10‐ol Content in Picea omorika,Pinus heldreichii,and Pinus peuce

The n‐alkane composition and the nonacosan‐10‐ol content in the needle cuticular waxes of Serbian spruce (Picea omorika), Bosnian pine (Pinus heldreichii), and Macedonian pine (Pinus peuce) were compared. The amount of nonacosan‐10‐ol in the needle waxes of P. omorika was higher than those in P. heldreichii and P. peuce. The range of n‐alkanes was also wider in P. omorika (C₁₈–C₃₅) than in P. heldreichii and P. peuce (C₁₈–C₃₃). The dominant n‐alkanes were C₂₉ in the needle waxes of P. omorika, C₂₃, C₂₇, and C₂₅ in those of P. heldreichii, and C₂₉, C₂₅, C₂₇, and C₂₃ in those of P. peuce. The waxes of P. omorika contained higher amounts of n‐alkanes C₂₉, C₃₁, and C₃₃, while those of P. heldreichii and P. peuce had higher contents of n‐alkanes C₂₁, C₂₂, C₂₃, C₂₄, and C₂₆. The principal component analysis of the contents of nine n‐alkanes showed a clear separation of the Serbian spruce populations from those of the two investigated pine species, which partially overlapped. The separation of the species was due to high contents of the n‐alkanes C₂₉ and C₃₁ (P. omorika), C₁₉, C₂₀, C₂₁, C₂₂, C₂₃, and C₂₄ (P. heldreichii), and C₂₈ (P. peuce). Cluster analysis also showed a clear separation between the P. omorika populations on one side and the P. heldreichii and P. peuce populations on the other side. The n‐alkane and terpene compositions are discussed in the light of their usefulness in chemotaxonomy as well as with regard to the biogeography and phylogeny of these rare and endemic conifers.     

Adaptational changes in cellular phospholipids and fatty acid composition of the food pathogen Listeria monocytogenes as a stress response to disinfectant sanitizer benzalkonium chloride

Aims: This study provides a first approach to observing the alterations of the cell membrane lipids in the adaptation response of Listeria monocytogenes to the sanitizer benzalkonium chloride. Methods and Results: A thorough investigation of the composition of polar and neutral lipids from L. monocytogenes grown when exposed to benzalkonium chloride is compared to cells optimally grown. The adaptation mechanism of L. monocytogenes in the presence of benzalkonium chloride caused (i) an increase in saturated‐chain fatty acids (mainly C_16:0 and C_18:0) and unsaturated fatty acids (mainly C_16:1 and C_18:1) at the expense of branched‐chain fatty acids (mainly C_a‐15:0 and C_a‐17:0) mainly because of neutral fatty acids; (ii) no alteration in the percentage of neutral and polar lipid content among total lipids; (iii) a decrease in lipid phosphorus and (iv) an obvious increase in the anionic phospholipids and a decrease in the amphiphilic phosphoaminolipid. Conclusions: These lipid changes could lead to decreased membrane fluidity and also to modifications of physicochemical properties of cell surface and thus changes in bacterial adhesion to abiotic surfaces. Significance and Impact of the Study: The adaptation and resistance of L. monocytogenes to disinfectants is able to change its physiology to allow growth in food‐processing plants. Understanding microbial stress response mechanisms would improve the effective use of disinfectants.     

Chemodiversity of Nonacosan‐10‐ol and n‐Alkanes in the Needle Wax of Pinus heldreichii

This is the first report of individual variability and population diversity of the contents of nonacosan‐10‐ol and n‐alkanes in the needle cuticular waxes of Bosnian pines originated from Montenegro, regarded as Pinus heldreichii var. leucodermis, and from Serbia, regarded as P. heldreichii var. pan?i?i. The amount of nonacosan‐10‐ol varied individually from 27.4 to 73.2% (55.5% in average), but differences between the four investigated populations were not statistically confirmed. The size of the n‐alkanes ranged from C₁₈ to C₃₃. The most abundant n‐alkanes were C₂₃, C₂₇, and C₂₅ (12.2, 11.2, and 10.8% in average, resp.). The carbon preference index (CPI) of the n‐alkanes ranged from 0.8 to 3.1 (1.6 in average), while the average chain length (ACL) ranged from 20.9 to 26.5 (24.4 in average). Long‐chain and mid‐chain n‐alkanes prevailed (49.6 and 37.9% in average, resp.). It was also found that the populations of P. heldreichii var. leucodermis had predominantly a narrower range of n‐alkanes (C₁₈? C₃₁) than the trees of the variety pan?i?i (C₁₈? C₃₃). Differences between the varieties were also significant for most of the other characteristics of the n‐alkane pattern (e.g., most abundant n‐alkanes, CPI, ACL, and relative proportion of short‐, mid‐, and long‐chain n‐alkanes). The principle component and cluster analyses of eleven n‐alkanes confirmed the significant diversity of these two varieties.     

Chemotaxonomic Considerations of the n‐Alkane Composition in Pinus heldreichii,P. nigra,and P. peuce

The n‐alkane composition in the leaf cuticular waxes of natural populations of Bosnian pine (Pinus heldreichii), Austrian pine (P. nigra), and Macedonian pine (P. peuce) was compared for the first time. The range of n‐alkanes was wider in P. nigra (C₁₆ – C₃₃) than in P. heldreichii and P. peuce (C₁₈ – C₃₃). Species also diverged in abundance and range of dominant n‐alkanes (P. heldreichii: C₂₃, C₂₇, and C₂₅; P. nigra: C₂₅, C₂₇, C₂₉, and C₂₃; P. peuce: C₂₉, C₂₅, C₂₇, and C₂₃). Multivariate statistical analyses (PCA, DA, and CA) generally pointed out separation of populations of P. nigra from populations of P. heldreichii and P. peuce (which were, to a greater or lesser extent, separated too). However, position of these species on the basis of n‐alkane composition was in accordance neither with infrageneric classification nor with recent molecular and terpene investigations.     

Comparison of growth and lipid composition in the green abalone, Haliotis fulgens, provided specific macroalgal diets

Lipid composition of abalone was examined over a one-year interval. A feeding trial was designed to cover a full reproductive cycle in young adult green abalone, Haliotis fulgens, consisting of five diet treatments: the macrophytic algal phaeophyte Egregia menziesii, rhodophyte Chondracanthus canaliculatus, chlorophyte Ulva lobata, a composite of the three algae and a starvation control. The lipid class, fatty acid, sterol and 1-O-alkyl glyceryl ether profiles were determined for foot, hepatopancreas/gonad tissues and larvae. The major fatty acids were 16:0, 18:0, 18:1(n-7)c, 18:1(n-9)c, 20:4(n-6), 20:5(n-3) and 22:5(n-3), as well as 14:0 for abalone fed brown and red algae. 4,8,12-Trimethyltridecanoic acid, derived from algae, was detected for the first time in H. fulgens (hepatopancreas complex, 1.2–13.9%; larvae, 0.5% of total fatty acids). Diacylglyceryl ethers were present in larvae (0.6% of total lipid). The major 1-O-alkyl glycerols were 16:0, 16:1 and 18:0. Additionally, 18:1(n-9) was a major component in hepatopancreas/gonad and larvae. The major sterol was cholesterol (96–100% of total sterols). Highest growth rates were linked to temperature and occurred in abalone fed the phaeophyte E. menziesii (43 μm·day^–1, 56 mg·day^–1 yearly mean), an alga containing the highest levels of C₂₀ polyunsaturated fatty acids and the highest ratio of 20:4(n-6) to 20:5(n-3). This study provides evidence of the influence of diet and temperature on seasonal changes in abalone lipid profiles, where diet is most strongly related to body mass and temperature to shell length. The allocation of lipids to specific tissues in green abalone clarifies their lipid metabolism. These results provide a basis for improving nutrition of abalone in mariculture through formulation of artificial feeds.     

Total lipid and fatty acid compositions of <Emphasis Type="Italic">Lertha sheppardi</Emphasis> (Neuroptera: Nemopteridae) during its main life stages

Total lipid and the fatty acid compositions of phospholipid and triacylglycerol fractions, prepared from eggs, 3^rd instars of larvae, pupae, male and female adults of Lertha sheppardi, were analyzed by gas chromatography and gas chromatography-mass spectrometry. The effect of diet (adults’ nutrition) on fatty acid composition of L. sheppardi adults was also investigated. Total lipid of L. sheppardi considerably increased in adults compared with immature stages. There was a significant decrease in total lipid level in larval stage in contrast with egg stage. Qualitative analysis revealed the presence of 14 fatty acids during all stages. The major components were C16 and C18 saturated and unsaturated components which are ubiquitous to most animal species. In addition to these components, one odd-chain (C17:0) and prostaglandin precursor fatty acids were found. The fatty acid profiles of phospholipids and triacylglycerols were substantially different. In phospholipid fraction, monounsaturated fatty acids were the major proportion of fatty acids in both sex of adults and pupae, whereas polyunsaturated fatty acids were the most dominant fatty acids in eggs and 3^rd instars. Results of triacylglycerol fraction revealed that fatty acid composition of eggs had higher level of C16:1, C18:0 and C18:3n-3 content than that of 3^rd instars and pupae, which suggests accumulation of energetic and structural reserve materials during embryonic development. At more advanced developmental stages, mainly in adult females, the amount of C16:1 increased once again, which may be related to the need for accumulation of sufficient energy and of carbon reservoir in the developing new vitellum. Percentages of C18:1 were significantly high in adult stages compared to other stages. These findings indicate that the accumulation and consumption of fatty acids fluctuate through different development stages. Diet did not effect the fatty acid composition of L. sheppardi adults.     

Effect of Conidiobolus coronatus on the Cuticular and Internal Lipid Composition of Tettigonia viridissima Males

Conidiobolus coronatus is an entomopathogenic fungus which has a potential as a biological control agent of insects. The cuticular and internal lipid composition of infected and noninfected Tettigonia viridissima males were analyzed by GC/MS. A total of 49 compounds were identified in the infected and noninfected males, including fatty acids, fatty acid methyl esters (FAMEs), n‐alkanes, alcohols, sterols, and other organic compounds. The most abundant components of the cuticular and internal lipids of the insects were fatty acids. After exposure to C. coronatus, the cuticular lipids of the T. viridissima males contained 17 free fatty acids from C(8) to C(22), while the cuticular lipids of the noninfected insects contained only 15 fatty acids from C(12) to C(24). The cuticular and internal lipids of both the infected and the noninfected males also contained five FAMEs from C(15) to C(19), seven n‐alkanes from C(25) to C(34), five alcohols from C(16) to C(25), five sterols, and the following six other organic compounds: azelaic acid, phenylacetic acid, glutaric acid, benzoic acid, sebacic acid, and glycerol. The compounds which were present only in the cuticular lipids of the infected males could be due to fungal infection.     

Effect of cell lipid composition on the formation of nonspecific antibiotic resistance in alkanotrophic rhodococci

The antibiotic resistance and lipid composition of rhodococci grown in rich organic media with gaseous or liquidn-alkanes were studied. Hydrocarbon-grown rhodococci exhibited an increased resistance to a wide range of antibiotics (aminoglycosides, linkosamides, macrolides, β-lactams, and aromatic compounds). The enhanced antibiotic resistance of rhodococci grown onn-alkanes correlated with an increased content of total cell lipids (up to 14–28%) and saturated straight-chain fatty acids (C_16:0, C_18:0, C_21:0) and was accompanied by the appearance of cardiolipin and phosphatidylglycerol in cells. These lipid compounds are supposed to promote the formation of nonspecific antibiotic resistance in rhodococci by decreasing the permeability of their cell envelope to antibiotics.     

Acyl Coenzyme A Synthetase Regulation: Putative Role in Long‐Chain Acyl Coenzyme A Partitioning

Objective: Long‐chain acyl coenzyme A synthetase (ACSL) converts free fatty acids (FFAs) into their metabolizable long‐chain acyl coenzyme A (LC‐CoA) derivatives that are essential for FFA conversion to CO₂, triglycerides, or complex lipids. ACSL‐1 is highly expressed in adipose tissue with broad substrate specificity. We tested the hypothesis that ACSL localization, and resulting local generation of LC‐CoA, regulates FFA partitioning. Research Methods and Procedures: These studies used cell fractionation of rat adipocytes to measure ACSL activity and mass and compared cells from young, mature, fed, fasted, and diabetic rats. Functional studies included measurement of FFA oxidation, complex lipid synthesis, and LC‐CoA levels. Results: High ACSL specific activity was expressed in the mitochondria/nuclei (M/N), high‐density microsomes (HDM), low‐density microsomes (LDM), and plasma membrane (PM) fractions. We show here that, during fasting, total FFA oxidation increased, and, although total ACSL activity decreased, a greater percentage of activity (43 ± 1.5%) was associated with the M/N fraction than in the fed state (23 ± 0.3%). In the fed state, more ACSL activity (34 ± 0.5%) was associated with the HDM than in the fasted state (25 ± 0.9%), concurrent with increased triglyceride formation from FFA. Insulin increased LC‐CoA and ACSL activity associated with the PM. The changes in ACSL activity in response to insulin were associated with only minor changes in mass as determined by Western blotting. Discussion: It is hypothesized that ACSL plays an important role in targeting FFA to specific metabolic pathways or acylation sites in the cell, thus acting as an important control mechanism in fuel partitioning. Localization of ACSL at the PM may serve to decrease FFA efflux and trap FFA within the cell as LC‐CoA.     

Distribution and Variability of n‐Alkanes in Epicuticular Waxes of Sedum Species from the Central Balkan Peninsula: Chemotaxonomic Importance

For the first time, the n‐alkane distribution and variability of the epicuticular waxes within 22 Sedum taxa was reported with focus on the chemotaxonomy of native Sedum representatives from the central Balkan Peninsula, compared to their relations with four other species of the Crassulaceae family. By GC/MS and GC‐FID identification and quantification, it was established that n‐alkanes C₂₇, C₂₉, C₃₁, C₃₃, and C₃₅ were the dominant constituents of the examined epicuticular wax samples. Applying multivariate statistical analyses including agglomerative hierarchical clustering (AHC) and principal component analysis (PCA), the relation according to the n‐alkane composition between the examined samples was established. It was shown that the n‐alkane variability of the central Balkan Sedum species was considerable and that n‐alkanes might not be very reliable taxonomic markers for these species.     

Comprehensive Characterisation of n‐Alkylresorcinols and Other Lipid Constituents of Mercurialis tomentosa L. from Alicante,Spain

Mercurialis tomentosa L. has been used in Spanish ethnomedicine. In the present study the first phytochemical characterisation of a lipid fraction from M. tomentosa was performed. The CHCl₃ extraction of aerial parts from M. tomentosa and GC/MS investigations revealed the occurrence of cuticular lipid and wax constituents, like long chain n‐alcohols and n‐aldehydes (C₂₂ – C₃₀), besides several aromatic constituents, i.e., phenylpropanoids and n‐alkylresorcinols. The latter were further purified by CC and analysed by LC/MSⁿ. In contrast to other Mercurialis species, i.e., M. annua, M. perennis, which exclusively contain 5‐n‐alkylresorcinols ( 1a  –  j , C_n), mainly 5‐n‐alkyl‐2‐methylresorcinols ( 2a  –  j , C_n*) with side chain lengths of C₁₅ – C₂₅ were found in M. tomentosa, in addition to 1a  –  j . Thus, the latter compounds may be utilised for analytical characterisation and authentication of M. tomentosa based on fingerprinting methods. For structure elucidation a novel facile total synthesis of one representative 5‐n‐alkyl‐2‐methylresorcinol homologue ( 2d , C₁₉*) was developed, starting with a Grignard reaction from a substituted benzoic acid chloride ( 19 ). The compound obtained by synthesis was identical to the natural product 2d in terms of its chromatographic and spectroscopic features. Futhermore, 2d exhibited satisfactory DPPH free radical scavenging activity (IC₅₀ = 37.8 μm ) when compared to trolox (IC₅₀ = 21.0 μm ), corroborating the antioxidant features of these amphipathic molecules.     

Effect of fasting on the composition of the fat body lipid of Dipetalogaster maximus, Triatoma infestans and Panstrongylus megistus (Hemiptera: Reduviidae)

Modifications in content and lipid composition induced by fasting were examined in fat bodies from adults of Triatominae, Dipetalogaster maximus, Triatoma infestans and Panstrongylus megistus. With fasting, total lipid stores dropped approximately 50% for T. infestans and more than 70% for P. megistus. Total lipids analyzed by thin layer chromatography and fractionated by column chromatography on Unisil showed triacylglycerols as the main component in the three species, although P. megistus showed high levels of diacylglycerols (31–46%). Cholesterol amounted to 8–15%. In diacylglycerol fractions, C_16:0, C_18:1 and C_18:0 fatty acids were detected; their ratio varied with species but it was not dependent on nutritional status. In triacylglycerol fractions C_18:1 fatty acid was the major component at different times (48–68%); the ratio of monounsaturated to saturated in this fraction was 1.3, 2.6 and 1.2 for D. maximus, T. infestans and P. megistus respectively. The remarkable drop in lipid stores without noticeable changes in their relative composition would suggest that all types of lipid are used at similar rates. The higher content of diacylglycerols in P. megistus may be associated with the better flight performance of this species. Accepted: 4 August 1998     

Ultrastructure,composition of neutral lipids and their fatty acids of Candida tropicalis strain D-2 mutants resistant to the polyene antibiotic nystatin

This report deals with data on the cell ultrastructure of Candida tropicals strain D-2 mutants resistant to the polyene antibiotic, nystatin, and with an analysis of the fractional composition of neutral lipids and their fatty acids. The ultrastructural organization of the mutant cells is characterized by thickening of the cell wall and formation of invaginations into the cytoplasm, the appearance of new formations, large vacuoles, and reduction of the system of mitochondrial cristae. Lipids of nys^r mutants differ from those of the nys^s variant in having a decreased content of steroids and some fractions of neutral lipids. Certain nys^r mutants manifest difference in the relative amounts of saturated and unsaturated fatty acids (C_16:0, C_16:1, C_18:0, C_18:1).