The molecular structure and multifunctionality of the cryptic plant polymer suberin.

Suberin, a plant polyester, consists of polyfunctional long-chain fatty acids and glycerol and is an intriguing candidate as a novel antimicrobial material. We purified suberin from cork using ionic-liquid catalysis during which the glycerol bonds that ensure the polymeric nature of suberin remained intact or were only partially cleaved-yielding the closest to a native configuration reported to date. The chemistry of suberin, both in situ (in cryogenically ground cork) and ex situ (ionic-liquid extracted), was elucidated using high-resolution one- and two-dimensional solution-state NMR analyses. Centrifugation was used to isolate suberin particles of distinct densities and their monomeric composition, assembly, and bactericidal effect, inter alia, were assessed. Analysis of the molecular structure of suberin revealed the relative abundance of linear aliphatic vs. acylglycerol esters, comprising all acylglycerol configurations and the amounts of total carbonyls (C[bond, double bond]O), free acid end groups (COOH), OH aliphatics, and OH aromatics. Suberin centrifuged fractions revealed generic physiochemical properties and monomeric composition â€‹and self-assemble into polygonal structures that display distinct degrees of compactness when lyophilized. Suberin particles-suberinsomes-display bactericidal activity against major human pathogenic bacteria. Fingerprinting the multifunctionality of complex (plant) polyesters such as suberin allows for the identification of novel polymer assemblies with significant value-added properties.

Antimicrobial activity of untypical lipid compounds in the cuticular and internal lipids of four fly species.

AIMS: This article describes the qualitative and quantitative analyses of untypical compounds in the cuticular and internal lipids of four dipteran species. For isolated compounds, antimicrobial activity against 18 reference strains of bacteria and fungi was determined. METHODS AND RESULTS: In this study, gas chromatography (GC) combined with mass spectrometry (GC-MS) was used to analyse the surface and internal compounds of four fly species. Seven untypical compounds from both pre-imaginal and imaginal stages of examined insects were identified. Azelaic acid (AA) was the most abundant, while phenylacetic and phenylpropionic acids occurred in lower concentration. Minor quantities of sebacic acid, 2-methyl-2-hydroxybutanoic acid, tocopherol acetate and trace amounts of 2,4-decadienal were also detected. Tocopherol acetate was found only in cuticular lipids of Musca domestica larvae. Each compound was tested against several species of fungi and bacteria by determining minimal inhibitory concentration (MIC). Human pathogenic fungi were also investigated. Phenylpropionic acid showed the greatest antifungal activity. Bacterial strains were insensitive to the presence of identified compounds, apart from 2,4-decadienal which strongly inhibited bacterial growth. CONCLUSIONS: This is the first time that the chemical composition and the antimicrobial activity of untypical compounds in the cuticular and internal lipids of four fly species has been analysed. SIGNIFICANCE AND IMPACT OF THE STUDY: Determination of untypical compounds and their antimicrobial activity can effectively contribute to the knowledge concerning insect defence mechanisms.

Cuticular and internal n-alkane composition of Lucilia sericata larvae, pupae, male and female imagines: application of HPLC-LLSD and GC/MS-SIM.

The composition of cuticular and internal n-alkanes in Lucilia sericata larvae, pupae, and male and female imagines were studied. The cuticular and internal lipid extracts were separated by HPLC-LLSD, after which the hydrocarbon fraction was identified by GC/MS in selected ion monitoring (SIM) and total ion current (TIC) modes. The cuticular lipids of the larvae contained seven n-alkanes from C23 to C31. The major n-alkane in L. sericata larvae was C29 (42.1%). The total cuticular n-alkane content in the cuticular lipids was 31.46 µg g-1 of the insect body. The internal lipids of L. sericata larvae contained five n-alkanes ranged from C25 to C31. The most abundant compound was C27 (61.71 µg g-1 of the insect body). Eighteen n-alkanes from C14 to C31 were identified in the cuticular lipids of the pupae. The most abundant n-alkanes ranged from C25 to C31; those with odd-numbered carbon chains were particularly abundant, the major one being C29:0 (59.5%). Traces of eight cuticular n-alkanes were present. The internal lipids of L. sericata pupae contained five n-alkanes, ranging from C25 to C31. The cuticular lipids of female imagines contained 17 n-alkanes from C12 to C30. Among the cuticular n-alkanes of females, C27 (47.5%) was the most abundant compound. Four n-alkanes, with only odd-numbered carbon chains, were identified in the internal lipids of females. The lipids from both sexes of L. sericata had similar n-alkane profiles. The cuticular lipids of adult males contained 16 n-alkanes ranging from C13 to C31. C27 (47.9%) was the most abundant cuticular n-alkanes in males. The same n-alkanes only with odd-numbered carbon chains and in smaller quantities of C27 (0.1%) were also identified in the internal lipids of males. The highest amounts of total cuticular n-alkanes were detected in males and females of L. sericata (330.4 and 158.93 µg g-1 of the insect body, respectively). The quantities of total cuticular alcohols in larvae and pupae were smaller (31.46 µg g-1 and 42.08 µg g-1, respectively). The internal n-alkane contents of larvae, pupae, and male and female imagines were significantly higher than the cuticular n-alkane contents (153.53, 99.60, 360.06 and 838.76 µg g-1 of the insect body, respectively).

Leaf cuticular n-alkanes as markers in the chemotaxonomy of the eggplant (Solanum melongena L.) and related species.

The complex of species formed by eggplant (Solanum melongena L.) and its wild and weedy relatives (mainly S. incanum L. and S. insanum L.) is characterised by an extreme morphological divergence that is not always associated with genetic variation. The taxonomy of so-called 'spiny Solanum' species (subgenus Leptostemonum) is therefore extremely unclear. Cultivated eggplant lacks resistance to pests that frequently occur among the wild forms and species. As these wild plants are a potential gene pool for improvement of eggplant cultivars, knowledge of the characteristics of taxonomic relations between plants of different origin is crucial. We suggest using the leaf cuticular n-alkane chain length distribution pattern as an alternative taxonomic marker for eggplant and related species. The results are in good agreement with current knowledge of the systematics of these plants; at the same time, the method developed here is useful for verifying plant identification based on morphological traits. Analysis of 13 eggplant cultivars, five accessions of S. incanum and two lines of S. macrocarpon enabled the intraspecific variation within eggplant to be assessed as low. There was wide variability among S. incanum accessions, probably because plants described as S. incanum are members of a number of different species. Some Asian accessions (sometimes described as S. insanum) were found to be almost identical to S. melongena, while a truly wild African S. incanum plant showed extensive similarity. The usefulness of the chemotaxonomic approach in dealing with the S. melongena-S. incanum complex is discussed.

Heterogeneous structure of O-Antigenic part of lipopolysaccharide of Salmonella Telaviv (Serogroup O:28) containing 3-Acetamido-3,6-dideoxy-D-glucopyranose.

The O-polysaccharide of Salmonella Telaviv was obtained by mild acid degradation of the lipopolysaccharide and studied by chemical methods (sugar and methylation analyses, Smith degradation, de-O-acetylation) and NMR spectroscopy. The structure of the O-polysaccharide was established. The repeating units that are proximal to the lipopolysaccharide core region mostly have a digalactose side chain and lack glucose, whereas those at the other end of the chain mostly do bear glucose but are devoid of the disaccharide side chain. This is the first structure established for the O-polysaccharide of a Salmonella serogroup O:28 (formerly M) strain characterized by subfactors O28(1) and O28(2). Knowledge of this structure and the structure of the O-polysaccharide of Salmonella Dakar (O28(1), O28(3)) established earlier is crucial for determination of the exact structures associated with subfactors O28(1), O28(2), and O28(3) and elucidation of the genetic basis of the close relationship between Escherichia coli O71 and S. enterica O:28 O-antigens.

Identification of ionic liquid breakdown products in an advanced oxidation system.

Commonly used alkylimidazolium ionic liquids are poorly to negligibly biodegradable, and some are toxic, with the potential to poison typical biological test systems. Therefore, when ionic liquids are present in technological wastewaters they could break through classical wastewater treatment systems into natural waters and become potentially persistent pollutants. A recent study investigating different advanced oxidation processes found that the H(2)O(2)/UV system degraded dissolved imidazolium ionic liquids with the greatest efficiency. In the present study, high performance liquid chromatography was coupled with electrospray mass spectrometry to separate, analyse and identify degradation products following the treatment of ionic liquid solutions with H(2)O(2) in the presence of UV irradiation. It was found that hydroxylation in short-chain entities occurred mainly within the ring moiety, whereas in the case of longer alkylated cations, oxidation of the alkyl chain yielded several products. The potential transformation products were identified structurally by MS/MS analysis and are discussed in the light of their putative toxicity and biodegradability.

Degradation of 1-butyl-3-methylimidazolium chloride ionic liquid in a Fenton-like system.

The study examined the usefulness of a Fenton-like system for the degradation of ionic liquid residues in water. The ionic liquid was oxidized in a dilute aqueous solution of 1-butyl-3-methylimidazolium chloride (bmimCl). The ionic liquid decomposes readily and rapidly in aqueous solution by chemical degradation in a Fenton-like system. Under chosen conditions the initial bmimCl solution was reduced by a factor of 0.973 within 90 min. Additional results showed that bmimCl degradations in a Fenton-like system in excess H2O2 could be interpreted as a combined oxidation-reduction mechanism. Preliminary investigations of the mechanism of such degradations have indicated that initial OH* radicals can attack any one of the three carbon atoms on the imidazolium ring. The intermediates of this reaction may be mono- di- or amino- carboxylic acids.

Assessing toxicity and biodegradation of novel, environmentally benign ionic liquids (1-alkoxymethyl-3-hydroxypyridinium chloride, saccharinate and acesulfamates) on cellular and molecular level.

Ionic liquids are widely studied as alternative solvents in organic synthesis and catalysis, in electrochemistry and the separation sciences; with their many applications they will soon be produced on an industrial scale. Available toxicological data of ionic liquids have already suggested initial guidelines for the conscious design of safer chemicals. In this study a new group of such redesigned ionic liquids-1-alkoxymethyl-3-hydroxypyridinium cations+acesulphamate, saccharinate and chloride anions-was assayed with respect to their inhibitory activity towards acetylcholinesterase and their cellular toxicity towards the IPC-81 rat promyelocytic leukaemia cell line: the acute biological activity of these compounds is very low. Effective concentrations lie in the millimole range, which is well above possible intracellular concentrations. Only the compounds with the longest alkoxymethyl chain inhibit the enzyme at effective concentrations that are one order of magnitude smaller. No significant differences are observed when the anion compartment in the enzymatic assay is varied. However, the cytotoxicity data show EC(50) for acesulphamates and saccharinates to be higher than the values for the chloride analogues. Also, a slight alkoxymethyl chain length effect on the overall cytotoxicity is discernible. The biodegradability of the 1-alkoxymethyl-3-hydroxypyridinium salts varies from 21% to 72% and depends on the type of anion the cation is linked with. It improves with lengthening alkyl chain, but only in the range from 4 to 11 carbon atoms.

Effective biotic elicitation of Ruta graveolens L. shoot cultures by lysates from Pectobacterium atrosepticum and Bacillus sp.

Growth of Ruta graveolens shoots was induced when Bacillus sp. cell lysates were added to the culture medium. Elicitation of coumarin by this lysate was also very effective; the concentrations of isopimpinelin, xanthotoxin and bergapten increased to 610, 2120 and 1460 microg g(-1) dry wt, respectively. It also had a significant effect on the production of psoralen and rutamarin (680 and 380 microg g(-1) dry wt) and induced the biosynthesis of chalepin, which was not detected in the control sample, up to 47 microg g(-1) dry wt With lysates of the Pectobacterium atrosepticum, their effect on growth was not so significant and had no effect on the induction of coumarin accumulation. But elicitation with this lysate was much more effective for inducing the production of furoquinolone alkaloids; the concentrations of gamma-fagarine, skimmianine, dictamnine and kokusaginine rose to 99, 680, 172 and 480 microg g(-1) dry wt, respectively.

Evaluation of the acute toxicity of perfluorinated carboxylic acids using eukaryotic cell lines, bacteria and enzymatic assays.

The acute biological activity of a homologous series of perfluorinated carboxylic acids - perfluorohexanoic acid (PFHxA), perfluoroheptanoic acid (PFHpA), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA) and perfluorodecanoic acid (PFDA) - was studied. To analyze the potential risk of the perfluorinated acids to humans and the environment, different in vitro toxicity test systems were employed. The cytotoxicity of the chemicals towards two different types of mammalian cell lines and one marine bacteria was investigated. The viability of cells from the promyelocytic leukemia rat cell line (IPC-81) and the rat glioma cell line (C6) was assayed calorimetrically with WST-1 reagent. The evaluation was combined with the Vibrio fischeri acute bioluminescence inhibition assay. The biological activity of the compounds was also determined at the molecular level with acetylcholinesterase and glutathione reductase inhibition assays. This is the first report of the effects of perfluorinated acids on the activity of purified enzymes. The results show these compounds have a very low acute biological activity. The observed effective concentrations lie in the millimole range, which is well above probable intracellular concentrations. A relationship was found between the toxicity of the perfluorinated carboxylic acids and the perfluorocarbon chain length: in every test system applied, the longer the perfluorocarbon chain, the more toxic was the acid. The lowest effective concentrations were thus recorded for perfluorononanoic and perfluorodecanoic acids.

Usefulness of pi...pi aromatic interactions in the selective separation and analysis of imidazolium and pyridinium ionic liquid cations.

Current research indicates that replacing organic solvents with room-temperature ionic liquids may lead to remarkable improvements in well-known processes. Ionic liquids have already been utilized as alternative solvents in organic synthesis and catalysis, and also in electrochemistry and in separation sciences. Their wide applicability will soon result in their production on an industrial scale. Therefore, analytical methods applicable to various matrices for product control and environmental monitoring will be very much in demand. In this study, the usefulness of pi...pi interactions between alkylimidazolium and pyridinium cations and the aromatic pi...pi active moiety of the stationary phase was investigated for the selective separation and analysis of some ionic liquids. With phenyl-bonded phases, very good separations of the cations were achieved. Special attention was paid to the short-chain hydrophilic entities, known to be poorly separated on conventional reversed-phase columns. Further, the nature of the interactions occurring in the system under study was investigated by varying the content of methanol/acetonitrile used as organic modifiers in the mobile phases. The analytical method developed here is simple and reproducible, and its quantitative analytical performance was excellent. The paper also discusses the applicability of the method for monitoring degraded cations of 1-butyl-3-methylimidazolium salts obtained in the Fenton process.

AMP deaminase in vitro inhibition by xenobiotics A potential molecular method for risk assessment of synthetic nitro- and polycyclic musks, imidazolium ionic liquids and N-glucopyranosyl ammonium salts.

The usefulness of in vitro AMP deaminase inhibition was examined as a potential molecular method in risk assessment of xenobiotics. The enzyme participates in the principal purine nucleotide interconversion and degradation pathways, and its absence caused perturbations in the cellular ATP pool. The compounds selected were synthetic musks with a known negative environmental impact and the toxicologically unknown ionic liquids and N-glucopyranosyl ammonium bromides, which have recently attracted much interest from the chemical and related industries. All the compounds tested demonstrated a dose-dependent inhibition of AMP deaminase activity. IC(50) ranged from 0.3µM for polycyclic musks to 500µM for N-glucopyranosyl trimethylammonium bromide. Analysis of Dixon plots showed the inhibition type for all the compounds to be noncompetitive. The results support the choice of such an assay for the prospective risk assessment of these compounds.

Evaluating the cytotoxicity of ionic liquids using human cell line HeLa.

This paper presents cytotoxicity data of selected imidazolium ionic liquids evaluated in vitro on the human tumor cell line HeLa. It was found that for 1-n-butyl-3-methylimidazolium entities the toxicity depends strongly on the associated anion; EC50 values are lowest for tetrafluoroborate. No direct dependence of the reduced effect concentration was found on elongating the short, methyl chain to ethyl or n-hexyl. Only for the ionic liquid with an n-decyl chain, the longest one studied, did higher hydrophobicity result in a EC50 one order of magnitude lower than that obtained with the n-butyl entity. The effect concentrations of imidazolium ionic liquids in the HeLa system used are lower than the values obtained for conventional organic solvents such as dichloromethane, toluene or xylene.

Recovery of astaxanthin from seafood wastewater utilizing fish scales waste.

The paper presents basic data on astaxanthin adsorption from fisheries wastewater to fish scales. This process has been proposed to be applicable in fisheries and shrimp waste management [Helgason, Recovery of compounds using a natural adsorbent, Patent WO 01/77230, 2001]. The innovative feature of the method is the application of a solid waste (fish scales) as a natural adsorbent for a carotenoid pigment (astaxanthin) from the seafood industry wastewater. The model investigations were performed with pure synthetic carotenoids to exclude the role of matrix in which astaxanthin is present in the wastewater. Under the experimental conditions used, the maximum loading capacity of astaxanthin onto the scales is 360 mg kg(-1) dry wt. Studies of the thus formed value added product indicated that drying causes significant loss of astaxanthin activity. Due to the effective filtration characteristics of the studied sorption material, we suggest the scale/astaxanthin sorption process to be suitable for treatment of wastewater from different industries.

Enhanced photo-degradation of contaminants in petroleum refinery wastewater.

In order to rise efficiency of the wastewater treatment in a refinery plant, several oxidation experiments were done, testing their applicability as an additional pretreatment method. The influence of treatment with low concentrations of H2O2 combined with stirring and UV light on degradation of organic compounds present in the refinery wastewater was studied. Oxidation of the total petroleum hydrocarbons occurs at relatively low concentrations of H2O2, additional UV irradiation slightly accelerates the process due to the increased formation of hydroxyl radicals. 1,2-dichloroethane and t-butyl methyl ether degrade in the similar manner and except for the lowest H2O2 concentration used (1.17 mM), the reduction after 24 h is total. The degradation rate for dichloromethane is the lowest one, depending both on hydrogen peroxide concentration and the presence of UV. Its maximum reduction of 83% was obtained using the highest applied peroxide concentration of 11.76 mM.