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Induction cues for tegument formation during the transformation of Schistosoma mansoni cercariae.

Adult schistosomes are parasitic blood flukes that have a continuous double lipid bilayered membrane surrounding the entire worm. This tegumental membrane is synthesised during invasion of the vertebrate host by free-swimming infectious forms called cercariae. As cercariae invade their final hosts they lose their tails and encounter a changing environment that includes altered temperature, sugar concentration and osmolarity. We have identified a glucose transporter protein designated SGTP4 that is found exclusively in the outer adult tegument and on membranous vesicles within the tegumental cytoplasm. By using immunofluorescence analysis to monitor the appearance and distribution of SGTP4 we can track the process of new tegumental membrane formation and examine the cues that trigger this developmental pathway. Cercariae in water do not transform their tegument while those incubated in rich medium do so rapidly. We have examined which of the many constituents of rich medium are responsible for triggering this transformation. Incubation in a solution of moderate osmolarity (120 mOsM PBS) is sufficient by itself to trigger tegument transformation, albeit at a slower rate relative to incubation in rich medium. Adding either glucose (to 100 mM) to the solution or increasing the temperature of incubation (from 22 degrees C to 37 degrees C) further increased the rate of tegument biogenesis. The introduction of glucose together with an increase in the incubation temperature further accelerated the process, suggesting that these factors act synergistically to promote transformation rates. The critical nature of osmolarity in inducing the process is highlighted by the fact that transformation proceeds as efficiently in 360 mOsM alone as it does in rich medium. While the fatty acids linolenic acid (cis-9, cis-12, cis-15-octadecatrienoic acid at 1 mM) and capric acid (Decanoic acid, at 0.1 mM) have both been proposed to stimulate tegumental transformation, we show that neither promotes the morphogenesis of a normal schistosomulum tegument. The schistosomicide praziquantel (to 1 mM) has no detectable effect on new tegument formation.

Animals↗

Comparison between cachaça and rum using pattern recognition methods.

The differentiation between cachaça and rum using analytical data referred to alcohols (methanol, propanol, isobutanol, and isopentanol), acetaldehyde, ethyl acetate, organic acids (octanoic acid, decanoic acid, and dodecanoic acid), metals (Al, Ca, Co, Cu, Cr, Fe, Mg, Mn, Ni, Na, and Zn), and polyphenols (protocatechuic acid, sinapaldehyde, syringaldehyde, ellagic acid, syringic acid, gallic acid, (-)-epicatechin, vanillic acid, vanillin, p-coumaric acid, coniferaldehyde, coniferyl alcohol, kaempferol, and quercetin) is described. The organic and metal analyte contents were determined in 18 cachaça and 21 rum samples using chromatographic methods (GC-MS, GC-FID, and HPLC-UV-vis) and inductively coupled plasma atomic emission spectrometry, respectively. The analytical data of the above compounds, when treated by principal component analysis, hierarchical cluster analysis, discriminant analysis, and K-nearest neighbor analysis, provide a very good discrimination between the two classes of beverages.

Acetaldehyde↗

Analysis of anal secretions from phlaeothripine thrips.

The anal secretions of 16 phlaeothripine thrips species (Thysanoptera: Phlaeothripidae) were studied, including a reinvestigation of three species previously reported. A total of 37 components were detected, including hydrocarbons, acetates, terpenes, carboxylic acids, a quinone, an aromatic compound, and a pyranone compound. The secretions of all species were composed of some of these components, with Xylaplothrips inquilinus possessing as many as 11 components. Of these components, (Z)-9-octadecene, (Z)-9-nonadecene, nonadecadiene, octanoic acid, decanoic acid, geranial, neral, alpha-pinene, beta-pinene, caryophyllene, 2-hydroxy-6-methylbenzaldehyde, and two unidentified monoterpenes [UK-I (M+136) and UK-II (M+168)] were detected for the first time. The chemicals were species-specific; four Liothrips species and three Holothrips species could be distinguished from each other and their congeners by the GC profiles of the ether extracts of their anal secretions. The anal secretions of gall-inducing thrips commonly contained terpenes. of which citral (a mixture of geranial and neral) and beta-acaridial repelled ants or had antifungal activity. The findings suggest that these terpenes play a defensive role and prevent galls from fungal infestation. 3-Butanoyl-4-hydroxy-6-methyl-2H-pyran-2-one, found from three Holothrips spp., caused paralysis in ants. Chemical analysis of anal secretion components is a useful method for the classification of tubuliferan species that are difficult to distinguish on the basis of morphological characters.

Adaptation, Physiological↗

The metabolism of n-decane by a Pseudomonas.

The growth of a Pseudomonas on n-decane was found to produce stearic acid, oleic acid, palmitic acid, palmitoleic acid, decanoic acid, octanoic acid, beta-hydroxydecanoic acid, beta-hydroxyoctanoic acid, beta-hydroxyhexanoic acid and beta-hydroxyadipic acid. Small amounts of n-decanamide and n-valeramide were also isolated. The effects of nitrogen and oxygen limitation on the formation of these products in continuous fermentations is reported.

Alkanes↗

Triglyceride synthesis by the small-intestinal epithelium of the pig, sheep and chicken.

1. A comparative study was made of triglyceride synthesis by the intestinal epithelium of pigs, sheep and chickens. In pig and chicken tissue both the glycerol 3-phosphate and the monoglyceride pathway of triglyceride synthesis were operative, but the former pathway predominated in sheep tissue. 2. The fatty acid specificity of the glycerol 3-phosphate pathway was studied in pig and sheep total-homogenate preparations. Maximum incorporation was obtained with myristic acid and palmitic acid under optimum conditions for each fatty acid. Lauric acid, myristic acid, oleic acid, linoleic acid and linolenic acid were inhibitory at concentrations above their optimum, but octanoic acid, decanoic acid, palmitic acid and stearic acid did not show this effect. 3. Subcellular fractionation located the glycerol 3-phosphate and monoglyceride pathways of triglyceride synthesis in the microsomes in all instances. Phosphatidate phosphohydrolase was associated with both the microsomes and the particle-free supernatant. 4. Glycerol 1-mono-oleate was incorporated into triglycerides to a greater extent than glycerol 1-mono-palmitate or glycerol 1-monostearate by microsomal preparations from pig and chicken. 5. A lipase specific for monoglycerides was detected in the particle-free supernatant of all the species examined.

Animals↗

Purification and kinetic mechanism of the major glutathione S-transferase from bovine brain.

The major glutathione S-transferase isoenzyme from bovine brain was isolated and purified approx. 500-fold. The enzyme has a pI of 7.39 +/- 0.02 and consists of two non-identical subunits having apparent Mr values of 22,000 and 24,000. The enzyme is uniformly distributed in brain, and kinetic data at pH 6.5 with 1-chloro-2,4-dinitrobenzene (CDNB) as substrate suggest a random rapid-equilibrium mechanism. The kinetics of inhibition by product, by GSH analogues and by NADH are consistent with the suggested mechanism and require inhibitor binding to several different enzyme forms. Long-chain fatty acids are excellent inhibitors of the enzyme, and values of 1nKi for hexanoic acid, octanoic acid, decanoic acid and lauric acid form a linear series when plotted as a function of alkyl chain length. A free-energy change of -1900 J/mol (-455 cal/mol) per CH2 unit is calculated for the contribution of hydrophobic binding energy to the inhibition constants. The turnover number of the purified enzyme dimer is approx. 3400/min. When compared with the second-order rate constant for the reaction between CDNB and GSH, the enzyme is providing a rate acceleration of about 1000-fold. The role of entropic contributions to this small rate acceleration is discussed.

Animals↗

Chemicals in laboratory room air stimulate olfactory neurons of female Bombyx mori.

Laboratory air contained odorants that elicited electrophysiological responses in female Bombyx mori antennae. Air entrainments on charcoal filters, extracted with CS(2) and subsequently with acetone, were analyzed by coupled gas chromatography (GC)-electroantennogram (EAG) as well as by GC-mass spectrometry. The CS(2) extract contained 12 EAG-active peaks from which benzaldehyde, octanal, limonene, 1,8-cineol, methyl benzoate, nonanal, decanal and geranyl acetone were identified. In the acetone extract we identified eight EAG-active peaks as phenol, nonanal, 2-ethylhexanoic acid, octanoic acid, benzoic acid, nonanoic acid, decanoic acid and dimethyl phthalate. The concentrations of benzoic acid and benzaldehyde present in laboratory air were determined. The origin of the substances and importance of such odorants in laboratory air for the interpretation of physiological experiments on the olfactory system is discussed.

Air↗

Effect of yeast hulls on stuck and sluggish wine fermentations: importance of the lipid component.

The effect of yeast hulls (yeast ghosts) on sluggish or stuck white wine fermentations was studied. The enhancing effect on yeast growth and fermentation rate displayed by the hulls was shown to be similar to the effect provided by lipid extract from the same hulls. Unsaturated fatty acids and sterols were incorporated into the yeast from lipid extracts during fermentation carried out under oxygen-limited conditions. Adsorption of toxic medium-chain fatty acid (decanoic acid) onto the yeast hulls took place through a dialysis membrane. However, when the hulls were placed inside a dialysis bag, the increase in yeast growth and fermentation rate seen when freely suspended hulls were used did not occur. Accordingly, the effect of yeast hulls in preventing stuck fermentations cannot be attributed only to the adsorption and consequent removal of medium-chain fatty acids from the juice.

Journal Article↗

The level of dietary protein and carbohydrate has a different effect on intestinal uptake of hexoses and lipids in rabbits with an ileal resection than in those with an intact intestinal tract.

Either high protein, low carbohydrate (HP) or low protein, high carbohydrate (LP) diets were fed for 6 weeks to rabbits with or without resection of the distal half of the small intestine. Control and resected rabbits fed HP consumed more food and gained more weight than the animals fed LP. The level of dietary protein has a different effect on intestinal transport in animals with an ileal resection than in those with an intact small intestine. With 0.5 mM glucose, the in vitro uptake in control rabbits was greater for the LP than HP diet but was unchanged in resected rabbits; uptake of 0.5 mM galactose and 3-O-methyl glucose was unaffected by HP and LP, whereas in rabbit uptake was lower in LP than HP. The uptake of 40 mM glucose was greater with the LP than HP diet in control rabbits, but lower with LP than HP in resected rabbits. In control rabbits, the uptake of aluric acid was lower on the LP than HP diet; the uptake of octanoic acid, decanoic acid, and cholesterol was unchanged; and the uptake of each fatty acid and cholesterol was greater in resected rabbits fed LP than HP. Feeding a low protein diet to animals with an ileal resection is associated with lower jejunal uptake of high concentrations of glucose, but the higher uptake of galactose and enhanced permeability to fatty acids result in superior weight gain. Thus, recommendations for alterations in dietary protein and carbohydrate levels following ileal resection must be made with the knowledge that these changes may influence intestinal transport function as well as body weight gain.

Animals↗

Peptide purification, complementary deoxyribonucleic acid (DNA) and genomic DNA cloning, and functional characterization of ghrelin in rainbow trout.

We have identified ghrelin from the stomach of rainbow trout. Four isoforms of ghrelin peptide were isolated: the C-terminal amidated type of rainbow trout ghrelin (rt ghrelin) composed of 24 amino acids (GSSFLSPSQKPQVRQGKGKPPRV-amide) is a basic form; des-VRQ-rt ghrelin, which deleted three amino acids (V13R14Q15) from rt ghrelin; and further two types of rt ghrelin that retained the glycine residue at the C terminus, rt ghrelin-Gly, and des-VRQ-rt ghrelin-Gly. The third serine residue was modified by octanoic acid, decanoic acid, or the unsaturated form of those fatty acids. In agreement with the isolated peptides, two cDNAs of different lengths were isolated. The rt ghrelin gene has five exons and four introns, and two different mRNA molecules are predicted to be produced by alternative splicing of the gene. A high level of ghrelin mRNA expression was detected in the stomach, and moderate levels were detected in the brain, hypothalamus, and intestinal tracts. Des-VRQ-rt ghrelin stimulated the release of GH in the rat in vivo. Furthermore, des-VRQ-rt ghrelin stimulated the release of GH, but not the release of prolactin and somatolactin in rainbow trout in vivo and in vitro. These results indicate that ghrelin is a novel GH secretagogue in rainbow trout that may affect somatic growth or osmoregulation through GH. Because ghrelin is expressed in various tissues other than stomach, it may play important role(s) in cellular function as a local regulator.

Amino Acid Sequence↗

Production of gamma-lactones by the brown-rot basidiomycete Piptoporus soloniensis.

A wild strain of brown-rot basidiomycete Piptoporus soloniensis produced a sweet flavor similar to tropical fruits in liquid cultures. The major and minor compounds were identified to be gamma-decalactone and gamma-octanolactone by gas chromatography-mass spectrometry analysis, respectively. The growth and production of gamma-decalactone by P. soloniensis in broth to which fatty acids had been added were investigated. The addition of 12-hydroxystearic acid and ricinoleic acid to the culture markedly enhanced the production of gamma-decalactone. On the other hand, addition of myristic acid, palmitic acid, stearic acid and oleic acid to the culture resulted in a higher production of gamma-octanolactone. The addition of hexanoic acid, octanoic acid, decanoic acid, lauric acid, linoleic acid and linolenic acid to the culture reduced the growth of P. soloniensis and production of gamma-decalactone and gamma-octanolactone. This strain accumulated oxalic acid in liquid culture and grew sufficiently under strongly acidic conditions.

Journal Article↗

Role of Yersinia intermedia and Pseudomonas putida in the development of a fruity off-flavour in pasteurized milk.

Analysis by gas chromatography-mass spectrometry of pasteurized milk with a fruity (pineapple like) off odour and a sour, rancid and soapy taste indicated the presence of concentrations at microg/ml levels of ethyl butanoate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, octanoic acid, decanoic acid and dodecanoic acid. The off-odour and taste were attributed to the presence of these compounds in the milk. Microbiological examination confirmed that the milk was also contaminated with a series of psychrotrophic bacteria including Yersinia intermedia, Pseudomonas putida and Rahnella aquatilis. Growth of isolates of these bacteria in UHT milk at 23 degrees C for 7 d showed that Yer. intermedia produced significant quantities of the C4-C12 alkanoic acids; Ps. putida produced only small quantities of these acids and Rah. aquatilis produced none. In addition, Yer. intermedia and Ps. putida also produced small but significant quantities of the corresponding ethyl esters. In milk inoculated with both Yer. intermedia and Ps. patida, the quantity of ethyl esters produced was greater than that found in cultures containing only one of the isolates. These studies indicated that Yer. intermedia was the principal source of the alkanoic acids in the tainted milk and that the major producer of the corresponding ethyl esters was Ps. patida. This is the first report that Yer. intermedia and Ps. putida can cause an off-odour or taste in dairy products.

Animals↗

Competition and coadsorption of di-acids and carboxylic acid solvents on HOPG.

The self-assemblies of di-acids HOOC-(CH(2))(n)-COOH (n = 20, 18, 16, 14, 12, 10) in three solvents hexanoic acid, octanoic acid, and decanoic acid on highly oriented pyrolytic graphite (HOPG) were studied with scanning tunneling microscopy (STM). In the solvent hexanoic acid, solvent molecules coadsorb with HOOC-(CH(2))(n)-COOH (n = 20, 18, 16) via formation of hydrogen bonds. The solvent octanoic acid coadsorbs with HOOC-(CH(2))(n)-COOH (n = 20, 18). Decanoic acid only coadsorbs with HOOC-(CH(2))(20)-COOH. In each solvent, the trend of coadsorption between solvent molecules and di-acid molecules decreases with decreasing chain-length of di-acid molecules. These systematic investigations suggest that coadsorption of solvent molecules with di-acid solute molecules is mainly dependent on the relative hydrogen-bond densities in the formed monolayer. This is consistent with the maximization of adsorption heat of the self-assembled monolayers of di-acids dissolved in solvents of carboxylic acids.

Journal Article↗

From decanoate micelles to decanoic acid/dodecylbenzenesulfonate vesicles.

Different aspects of mixtures of decanoic acid and sodium decanoate were investigated in aqueous solution up to a total concentration of 300 mM. Depending on the ratio of ionized to nonionized decanoic acid, micelles or vesicles form above the critical concentrations of micelle (cmc) or the critical concentration for vesicle formation (cvc). The micelles and the vesicles are always present together with nonmicellized or nonvesiculized decanoate. The latter was determined for different total concentrations. On the basis of titration curves, by application of the Gibbs phase rule, and on the basis of differential scanning calorimetry measurements and an electron microscopy analysis, the pH region within which vesicles exist was identified (pH 6.8-7.8). At pH 7.0, the concentration of nonvesiculized decanoate is approximately 20 mM. Decanoic acid/decanoate vesicles can be sized down by the extrusion technique to form stable and mainly unilamellar vesicles with a mean diameter of less than 100 nm. By coaddition of an equimolar amount of sodium dodecylbenzenesulfonate (SDBS) to decanoic acid, vesicles also formed below pH 6.8. These mixed vesicles were investigated as potential templates for the peroxidase-catalyzed polymerization of aniline at pH 4.3. Furthermore, decanaote micelles (at pH 11.0) were applied as reaction modifiers for the simultaneous competitive alkaline hydrolysis of p-nitrophenylacetate and fluorescein diacetate. While the rate of hydrolysis of fluorescein diacetate is slowed considerably in the presence of the micelles in comparison with the micelle-free system, the rate of hydrolysis of p-nitrophenylacetate remains almost unaffected.

Journal Article↗

In vitro studies on the oxidation of medium-chain dicarboxylic acids in rat liver.

The degradation of medium-chained dicarboxylic (DC) acids was investigated on purified mitochondria and peroxisomes. Intact organelles were incubated with dodecanedioic acid (DC12), suberic acid (DC8) and adipic acid (DC6), and the production of lower-chained DC-acids and of acetyl-CoA + acetyl-carnitine was monitored. It was shown, that intact peroxisomes could beta-oxidize DC12, DC10, and DC8 at least as far as DC6, while intact mitochondria readily beta-oxidized DC12, and DC10 as far as succinic acid. DC8 and DC6 were not oxidized by intact mitochondria when these two acids were presented externally to the intact organelle. When they were formed intramitochondrially from DC12 and DC10, both DC8 and DC6 were, however, to a great extent beta-oxidized as far as succinic acid. The major reason for this difference between mitochondrial oxidation of externally and internally located DC8 and DC6 seems to be an inability to transport these two acids through the mitochondrial membrane. For DC12 and DC10, the mitochondrial transport systems, which were indicated to be identical to the systems used by the corresponding monocarboxylic acids, were found to be rate-limiting in the beta-oxidation of these acids. A contributing factor to the undetectable beta-oxidation of externally located DC8 and DC6 may also be, that the Km values of DC8-CoA (460 +/- 70 mumol/l) and DC6-CoA (980 +/- 90 mumol/l) towards the acyl-CoA dehydrogenases are very high. These results imply that very high concentrations of intermediates are created intramitochondrially during beta-oxidation, concentrations which are probably only formed through formation of DC8-CoA and DC6-CoA from longer DC-acids and not by transport from outside the mitochondria. The data presented thus for the first time give evidence to a pathway for medium-chained monocarboxylic acids (especially lauric acid and decanoic acid) through cytosolic omega-oxidation followed by activation, transport over the mitochondrial membrane and beta-oxidation to succinic acid.

Acetyl Coenzyme A↗

Use of a fluorescent radiolabeled triacylglycerol as a substrate for lipoprotein lipase and hepatic triglyceride lipase.

A fluorescent radiolabeled triacylglycerol has been synthesized by using a fluorescent fatty acid (pyrene decanoic acid) and a radiolabeled oleic acid. This analog of the natural substrate, 1(3)pyrene decanoic-2,3 (1,2)-dioleoyl-sn-glycerol, has been tested as substrate for determining lipoprotein lipase and hepatic triacylglycerol lipase activities in post-heparin plasma. Optimal conditions for the determination of the two post-heparin plasma lipases were similar to those using radiolabeled triolein. Using this substrate, both post-heparin lipases exhibited their characteristic properties (pH optimum and effect of inhibitors) and attacked external ester bonds (1 or 3) containing pyrene decanoic and oleic acids at a similar rate.

Animals↗

General (medium-chain) acyl-CoA dehydrogenase deficiency (non-ketotic dicarboxylic aciduria): quantitative urinary excretion pattern of 23 biologically significant organic acids in three cases.

Urinary analysis of the pattern of 23 organic acid metabolites derived from fatty acids in three patients with general (medium-chain) acyl-CoA dehydrogenase deficiency was performed. Although there exist quantitative differences in the excreted amounts of the different metabolites in the three patients the qualitative picture was the same. The excretion of adipic, suberic and sebacic acids was substantial, whereas that of dodecanedioic acid was within or just above control limit. The monounsaturated C6-C10-dicarboxylic acid excretion was only marginally or not increased. 5-OH-hexanoic acid and hexanoylglycine were excreted in excessive amounts, whereas 7-OH-octanoic acid, 9-OH-decanoic acid, octanoylglycine and decanoylglycine were excreted in limited amounts. The excreted amounts of 6-OH-hexanoic, 8-OH-octanoic and 10-OH-decanoic acids were not or only marginally elevated compared to controls. In one of the patients the excretion of ethylmalonic and methylsuccinic acids was enhanced, whereas the excretion of these two acids in the two other patients was comparable to that in controls. The urinary excretion of hexanoic, octanoic, decanoic and dodecanoic acids was just a little above the control limit, whereas the esterified hexanoic and octanoic acids were excreted in appreciable amounts. It is argued that the microsomal omega- and omega-1-oxidation systems are involved in the dicarboxylic and omega-1-OH-monocarboxylic acids formation at C10 and C12 level and that the C8-C6-dicarboxylic and omega-1-OH-monocarboxylic acids are formed from higher chained acids by beta-oxidation in both mitochondria and peroxisomes.

Acyl-CoA Dehydrogenase↗

Beta-oxidation of C-6-C-10 fatty acids in rat liver homogenates measured by selected ion monitoring: effects of cyanide and clofibrate.

The beta-oxidation of C-6-C-10-fatty acids/acyl CoA/acylcarnitines in whole rat liver homogenates was determined by specific and simultaneous measurements of the C-6-C-10-fatty acids, i.e. hexanoic, octanoic and decanoic acids, in hydrolysed homogenates in relation to time in assays incubated with the above-mentioned substrates. Measurements were performed by a combined gas chromatographic mass spectrometric technique, i.e. selected ion monitoring. The rate of beta-oxidation of the C-6-C-10-fatty acids/acyl CoA/acylcarnitines were registered as the consumption rate of the added substrate. The conversion to the C-2-shortened beta-oxidation products was illustrated simultaneously. The rate of beta-oxidation of C-6-C-10-fatty acids was several times higher in homogenates from clofibrate-treated rats than in control rats, both in the presence and absence of 2.0 mmol l-1 cyanide. Cyanide caused a minor but significant decrease in the beta-oxidation rate in both control and clofibrate-treated rats. No differences were found between the beta-oxidation of decanoic acid and decanoyl CoA in homogenates from clofibrate-treated rats, whereas the degree of beta-oxidation of DL-decanoylcarnitine was halved compared with decanoic acid and decanoyl CoA.

Animals↗