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Differential inhibition of neuronal Na+-Ca2+ exchange versus store-operated Ca2+ channels by volatile anesthetics in pheochromocytoma (PC12) cells.

BACKGROUND: Ca2+ influx is a key component of neuronal intracellular Ca2+ ([Ca2+]i) regulation. The authors hypothesized that volatile anesthetic inhibition of neuronal activity is mediated by inhibition of Ca2+ influx via two major mechanisms: plasma membrane Na+-Ca2+ exchange (NCX) and the novel mechanism of Ca2+ influx triggered by endoplasmic reticulum Ca2+ depletion (store-operated Ca2+ channels [SOCCs]). METHODS: Differentiated rat pheochromocytoma cells loaded with the Ca2+ indicator fura-2 were Na+-loaded with 0 Ca2+, 145 mm Na+ Tyrode's and 5 microm cyclopiazonic acid plus 10 microm ryanodine (functionally isolating plasma membrane). Influx-mode NCX was rapidly reactivated by 0 Na+ and 2.5 mm Ca2+. The protocol was repeated in the presence of volatile anesthetics (0.5-1.5 minimum alveolar concentration [MAC] halothane, isoflurane, or sevoflurane) or other drugs to characterize NCX. To examine SOCCs, endoplasmic reticulum Ca2+ was depleted by cyclopiazonic acid in 0 extracellular Ca2+, and Ca2+ influx was triggered by rapid reintroduction of extracellular Ca2+. The protocol was repeated in the presence of anesthetics or other drugs to characterize SOCCs. RESULTS: Influx via NCX was not inhibited by voltage-gated Ca2+ channel blockers but was sensitive to NCX inhibitors. Halothane and isoflurane (0.5-1.5 MAC) significantly inhibited NCX (P < 0.05; paired comparisons), whereas sevoflurane at less than 1.5 MAC did not inhibit NCX. SOCC-mediated Ca2+ influx was insensitive to a variety of Ca2+ channel blockers but was inhibited by Ni2+. Such influx was sensitive only to halothane at greater than 1 MAC but not isoflurane or sevoflurane. CONCLUSIONS: These data indicate that volatile anesthetics, especially halothane and isoflurane, interfere with neuronal [Ca2+]i regulation by inhibiting NCX but not SOCC-mediated Ca2+ influx (except high concentrations of halothane).

Anesthetics, Inhalation↗

Volatiles from potato plants infected with potato leafroll virus attract and arrest the virus vector, Myzus persicae (Homoptera: Aphididae).

The influence of viral disease symptoms on the behaviour of virus vectors has implications for disease epidemiology. Here we show that previously reported preferential colonization of potatoes infected by potato leafroll virus (genus Polerovirus) (luteovirus) (PLRV) by alatae of Myzus persicae, the principal aphid vector of PLRV, is influenced by volatile emissions from PLRV-infected plants. First, in our bioassays both differential immigration and emigration were involved in preferential colonization by aphids of PLRV-infected plants. Second, M. persicae apterae aggregated preferentially, on screening above leaflets of PLRV-infected potatoes as compared with leaflets from uninfected plants, or from plants infected with potato virus X (PVX) or potato virus Y (PVY). Third, the aphids aggregated preferentially on screening over leaflet models treated with volatiles collected from PLRV-infected plants as compared with those collected from uninfected plants. The specific cues eliciting the aphid responses were not determined, but differences between headspace volatiles of infected and uninfected plants suggest possible ones.

Animals↗

Volatile antimicrobials from Muscodor albus, a novel endophytic fungus.

Muscodor albus is a recently described endophytic fungus obtained from small limbs of Cinnamomum zeylanicum (cinnamon tree). This xylariaceaous fungus effectively inhibits and kills certain other fungi, and bacteria, by virtue of a mixture of volatile compounds that it produces. The majority of these compounds were identified by gas chromatography/mass spectrometry and then made into an artificial mixture that mimicked the antibiotic effects of the mixture of volatile compounds given off by the fungus. Each of the five classes of volatile compounds produced by the fungus (alcohols, esters, ketones, acids and lipids) had some inhibitory effect against the test fungi and bacteria, but none was lethal. However, collectively they acted synergistically to kill a broad range of plant- and human-pathogenic fungi and bacteria. The most effective class of inhibitory compounds was the esters, of which 1-butanol, 3-methyl-, acetate was the most active biologically. This report describes the ecological implications and potential practical benefits of the 'mycofumigation' effects of M. albus.

Aldehydes↗

Nitrogen deficiency increases volicitin-induced volatile emission, jasmonic acid accumulation, and ethylene sensitivity in maize.

Insect herbivore-induced plant volatile emission and the subsequent attraction of natural enemies is facilitated by fatty acid-amino acid conjugate (FAC) elicitors, such as volicitin [N-(17-hydroxylinolenoyl)-L-glutamine], present in caterpillar oral secretions. Insect-induced jasmonic acid (JA) and ethylene (E) are believed to mediate the magnitude of this variable response. In maize (Zea mays) seedlings, we examined the interaction of volicitin, JA, and E on the induction of volatile emission at different levels of nitrogen (N) availability that are known to influence E sensitivity. N availability and volicitin-induced sesquiterpene emission are inversely related as maximal responses were elicited in N-deficient plants. Plants with low N availability demonstrated similar volatile responses to volicitin (1 nmol plant(-1)) and JA (100 nmol plant(-1)). In contrast, plants with medium N availability released much lower amounts of volicitin-induced sesquiterpenes compared with JA, suggesting an alteration in volicitin-induced JA levels. As predicted, low N plants exhibited greater sustained increases in wound- and volicitin-induced JA levels compared with medium N plants. N availability also altered volicitin-E interactions. In low N plants, E synergized volicitin-induced sesquiterpene and indole emission 4- to 12-fold, with significant interactions first detected at 10 nL L(-1) E. Medium N plants demonstrated greatly reduced volicitin-E interactions. Volicitin-induced sesquiterpene emission was increased by E and was decreased by pretreatment the E perception inhibitor 1-methylcyclopropene without alteration in volicitin-induced JA levels. N availability influences plant responses to insect-derived elicitors through changes in E sensitivity and E-independent JA kinetics.

Animals↗

Selenium assimilation and volatilization from dimethylselenoniopropionate by Indian mustard.

Earlier work from our laboratory on Indian mustard (Brassica juncea L.) identified the following rate-limiting steps for the assimilation and volatilization of selenate to dimethyl selenide (DMSe): (a) uptake of selenate, (b) activation of selenate by ATP sulfurylase, and (b) conversion of selenomethionine (SeMet) to DMSe. The present study showed that shoots of selenate-treated plants accumulated very low concentrations of dimethylselenoniopropionate (DMSeP). Selenonium compounds such as DMSeP are the most likely precursors of DMSe. DMSeP-supplied plants volatilized Se at a rate 113 times higher than that measured from plants supplied with selenate, 38 times higher than from selenite, and six times higher than from SeMet. The conversion of SeMet to selenonium compounds such as DMSeP is likely to be rate-limiting for DMSe production, but not the formation of DMSe from DMSeP because DMSeP was the rate of Se volatilization from faster than from SeMet and SeMet (but no DMSeP) accumulated in selenite- or SeMet-supplied wild-type plants and in selenate-supplied ATP-sulfurylase transgenic plants. DMSeP-supplied plants absorbed the most Se from the external medium compared with plants supplied with SeMet, selenate, or selenite; they also accumulated more Se in shoots than in roots as an unknown organic compound resembling a mixture of DMSeP and selenocysteine.

Brassica↗

Ion channel-forming alamethicin is a potent elicitor of volatile biosynthesis and tendril coiling. Cross talk between jasmonate and salicylate signaling in lima bean.

Alamethicin (ALA), a voltage-gated, ion channel-forming peptide mixture from Trichoderma viride, is a potent elicitor of the biosynthesis of volatile compounds in lima bean (Phaseolus lunatus). Unlike elicitation with jasmonic acid or herbivore damage, the blend of substances emitted comprises only the two homoterpenes, 4,11-dimethylnona-1,3,7-triene and 4,8,12-trimethyltrideca-1,3,7,11-tetraene, and methyl salicylate. Inhibition of octadecanoid signaling by aristolochic acid and phenidone as well as mass spectrometric analysis of endogenous jasmonate demonstrate that ALA induces the biosynthesis of volatile compounds principally via the octadecanoid-signaling pathway (20-fold increase of jasmonic acid). ALA also up-regulates salicylate biosynthesis, and the time course of the production of endogenous salicylate correlates well with the appearance of the methyl ester in the gas phase. The massive up-regulation of the SA-pathway (90-fold) interferes with steps in the biosynthetic pathway downstream of 12-oxophytodienoic acid and thereby reduces the pattern of emitted volatiles to compounds previously shown to be induced by early octadecanoids. ALA also induces tendril coiling in various species like Pisum, Lathyrus, and Bryonia, but the response appears to be independent from octadecanoid biosynthesis, because inhibitors of lipoxygenase and phospholipase A(2) do not prevent the coiling reaction.

Alamethicin↗

Effects of volatile anesthetics on cardiac calcium channels.

In order to investigate how volatile anesthetics affect cardiac calcium channels, the effects of halothane, enflurane, and isoflurane on the specific binding of [3H]-nitrendipine to bovine heart sarcolemmal membranes were studied. All three anesthetics added in liquid form inhibited [3H]-nitrendipine binding in a dose-dependent manner, and more interestingly, the order of inhibition by these volatile anesthetics roughly followed that of their anesthetic potencies. The partial pressures, calculated using the gas/water partition coefficients of halothane, enflurane, and isoflurane which inhibited [3H]-nitrendipine binding by 30% at 37 degrees C were about 1.48 x 10(-2) atm. (1.48%), 4.89 x 10(-2) atm. (4.89%) and 2.76 x 10(-2) atm. (2.76%), respectively. One mmol/l halothane altered not only the maximal binding (Bmax) from 189 f mol/mg protein to 136 f mol/mg protein, but also the dissociation constant (Kd) from 0.074 nmol/l to 0.18 nmol/l. Halothane was also added to the reaction mixture in the gaseous form with air. The partial pressure of halothane needed to bring about 30% inhibition was 0.82 x 10(-2) (0.82%), a value almost similar to that for halothane added in the liquid form. These results indicate that all three volatile anesthetics have direct effects on cardiac calcium channels, and that the magnitude of the effects depends on their anesthetic potencies.

Animals↗

The effects of volatile microbial secondary metabolites on protein synthesis in Serpula lacrymans.

The effects of volatile secondary metabolites produced by Trichoderma pseudokoningii, Trichoderma viride and Trichoderma aureoviride on growth rate and protein synthesis in two Serpula lacrymans isolates were investigated. Mycelial growth was affected to differing degrees, depending on the specific interactive microbial couplet involved. Protein synthesis by both S. lacrymans (Forfar) and S. lacrymans (H28) was affected by the volatile secondary metabolites of T. aureoviride and T. viride, but not by those of T. pseudokoningii. Mycelial growth and the original pattern of protein synthesis resumed when the antagonists were removed. It is probable that volatile secondary metabolites have played an important role during the evolution of microorganisms in the context of community, population and functional dynamics.

Basidiomycota↗

Protective effect of volatile oil, alcoholic and aqueous extracts of Origanum majorana on lead acetate toxicity in mice.

Natural dietary antioxidants are extensively studied for their ability to protect cells from miscellaneous damages. Origanum majorana L., Lamiaceae, is a potent antioxidant. The effect of administration of O. majorana (volatile oil, alcoholic and aqueous extracts) on oral administration of lead acetate in the diet of mice at concentration 0.5% (W/W) for one month were studied by measuring serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), urea and creatinine, histopathological changes of the liver and kidney and genotoxicity including, rate of micronucleus and chromosomal aberrations in bone marrow cells. Mice were treated with the 3 different forms of O. majorana, one month before and maintained with lead acetate administration. The 3 forms of O. majorana induced a significant decrease in serum activities of transaminases (AST & ALT), ALP, urea and creatinine and improved the liver and kidney histology in comparison with lead acetate treated group. Alcoholic extracts of O. majorana significantly reduced the rate of micronucleus, number of aberrant cells and different kinds of chromosomal aberrations. Volatile oil extract significantly reduced the rate of micronucleus and chromosomal fragments. Aqueous extract and volatile oil also of O. majorana significantly reduced number of gaps, ring chromosome and stickiness. It could be concluded that O. majorana plays an important role in ameliorating liver and kidney functions and genotoxicity induced by lead toxicity.

Animals↗

Effects of volatile constituents of rosemary extract on lung inflammation induced by diesel exhaust particles.

Epidemiological and experimental studies have implicated that diesel exhaust particles are involved in increases in morbidity and mortality from lung diseases. Recently, we have demonstrated that rosmarinic acid, a polyphenolic liquid component in perilla, inhibits lung inflammation induced by diesel exhaust particles in vivo, partly through its antioxidative property. We have also shown the antioxidative activities of volatile constituents of rosemary extract, the gaseous component in perilla, in vitro. The purpose of this study was to evaluate the effects of intratracheal administration of volatile rosemary extract on lung inflammation induced by diesel exhaust particles. ICR mice were treated with intratracheal administration of volatile rosemary extract before intratracheal exposure to diesel exhaust particles. Twenty-four hr later, diesel exhaust particles exposure elicited lung inflammation characterized by the infiltration of neutrophils and eosinophils, which was confirmed by cellular profile of bronchoalveolar lavage fluid and histological examination. Diesel exhaust particles enhanced the protein expressions of interleukin-1beta, macrophage inflammatory protein-1alpha, macrophage chemoattractant protein-1, and keratinocyte chemoattractant in the lung. Pretreatment with rosemary extract significantly inhibited the diesel exhaust particles-induced lung inflammation. Rosemary extract treatment also suppressed the diesel exhaust particles-enhanced lung expression of macrophage inflammatory protein-1alpha, macrophage chemoattractant protein-1, and keratinocyte chemoattractant. These results suggest that intratracheal administration of rosemary extract can prevent lung inflammation induced by diesel exhaust particles. The preventive effect is mediated, at least partly, through the inhibition of the enhanced lung expressions of macrophage inflammatory protein-1alpha, macrophage chemoattractant protein-1, and keratinocyte chemoattractants.

Administration, Inhalation↗

Genetic determinants of volatile-thiol release by Saccharomyces cerevisiae during wine fermentation.

Volatile thiols, particularly 4-mercapto-4-methylpentan-2-one (4MMP), make an important contribution to the aroma of wine. During wine fermentation, Saccharomyces cerevisiae mediates the cleavage of a nonvolatile cysteinylated precursor in grape juice (Cys-4MMP) to release the volatile thiol 4MMP. Carbon-sulfur lyases are anticipated to be involved in this reaction. To establish the mechanism of 4MMP release and to develop strains that modulate its release, the effect of deleting genes encoding putative yeast carbon-sulfur lyases on the cleavage of Cys-4MMP was tested. The results led to the identification of four genes that influence the release of the volatile thiol 4MMP in a laboratory strain, indicating that the mechanism of release involves multiple genes. Deletion of the same genes from a homozygous derivative of the commercial wine yeast VL3 confirmed the importance of these genes in affecting 4MMP release. A strain deleted in a putative carbon-sulfur lyase gene, YAL012W, produced a second sulfur compound at significantly higher concentrations than those produced by the wild-type strain. Using mass spectrometry, this compound was identified as 2-methyltetrathiophen-3-one (MTHT), which was previously shown to contribute to wine aroma but was of unknown biosynthetic origin. The formation of MTHT in YAL012W deletion strains indicates a yeast biosynthetic origin of MTHT. The results demonstrate that the mechanism of synthesis of yeast-derived wine aroma components, even those present in small concentrations, can be investigated using genetic screens.

Fermentation↗

Changes in the mouse intestinal microflora during weaning: role of volatile fatty acids.

The influence of volatile fatty acids on the ecology of the bacterial flora of the mouse intestinal tract has been studied in three situations where large fluctuations in the composition of the microflora have been observed. Young mice were shown to ingest solid food particles when 11 days old; this correlated with the appearance of strictly anaerobic fusiform bacilli in the intestinal lumen and a 10,000-fold decrease in numbers of coliform bacilli. Over the same period, volatile fatty acids were shown by gas-liquid chromatography to appear in the intestinal content. It is suggested that the fusiform bacilli are responsible for the presence of the volatile acids (especially butyric acid) which exert an inhibitory effect on the coliform bacteria, resulting in the decline in numbers. When germ-free mice are placed in a specific pathogen-free mouse colony, changes in the intestinal flora occurred which were similar to those observed in the young mice approaching weaning. Once again, the decline in the coliform population correlated with the appearance of significant levels of butyric acid in the large intestine. In a further series of experiments, mice were fed penicillin and levels of the intestinal fatty acids were measured. The antibiotic eliminated the anaerobic fusiforms from the intestine, resulting in the disappearance of significant levels of butyric acid and a million-fold increase in the numbers of coliform bacilli.

Age Factors↗

Volatile fatty acids in the faeces of patients in 'germ-free' isolation.

Faecal volatile fatty acids represent the end products of the metabolism of the anaerobic flora of the large bowel. The excretion of these volatile acids has been investigated in five leukaemic patients maintained in plastic ;germ-free' isolators. Under ;isolator' conditions there is a pronounced fall in volatile fatty acid excretion. The possibility that the measurement of these acids may be used to monitor anaerobic overgrowth and recolonization in these patients is discussed.

Adult↗

Analysis by gas liquid chromatography of production of volatile fatty acids by anaerobic bacteria grown on solid medium.

Volatile fatty acids produced in Robertson's cooked meat medium by a range of clinically relevant anaerobes were compared by gas liquid chromatography with those produced in blood agar. The same volatile fatty acid profiles were obtained in both media, although the concentration of acids was lower in blood agar. We conclude that detection of volatile fatty acids from a pure culture of an organism on solid medium is practicable and offers advantages over the conventional technique.

Acetates↗

Effects of wheat bran and porridge oats on hepatic portal venous volatile fatty acids in the pig.

Adult male pigs (40-60 kg of body weight) of the Kangaroo Island strain were surgically implanted with chronic indwelling hepatic portal venous cannulae. After a 24-hour fast the animals were given meals containing 500 g of either wheat bran or porridge oats and 200 g of sucrose and 2 litres of milk. With both cereal preparations plasma volatile fatty acids rose in the hepatic portal vein but the increase was significantly greater with wheat bran. Omission of sucrose and milk did not alter the response to porridge oats but diminished the response to wheat bran. These changes in plasma volatile fatty acids were unaffected by prior cooking of the cereals with hot water. With all test meals acetate and propionate were the major acids found, with butyrate contributing less than 8% of the total. This compositional profile was also found when the pigs were fed a commercial ration. The absence of butyrate differed from observations in the rat and reflected low concentrations of this acid in large bowel digesta. The difference in the response of the concentration of volatile fatty acids to feeding porridge oats and wheat bran in the pig was also the reverse of that found in the rat. These species differences may be of significance in relation to the choice of animal models for human fibre metabolism.

Animals↗

Two in vitro models for gas-phase exposure to volatile compounds.

Two experimental models suitable for the screening of volatile compounds were set up. The first consisted of a glass-chamber slide with eight wells, one carrying the test compound, and the others carrying cells in monolayers. In the second model, the cells were cultured in a glass Petri dish, and the test compound was poured onto a filter lying on a glass cover-slip, supported by a metal ring. Four plant volatiles [carvacrol, S-(+)- carvone, thymol and decanal] and one essential oil (caraway oil) were chosen as test compounds. The toxicity rankings obtained with the two models were compared with that obtained in a previous study performed with the same compounds under conventional culture conditions. Differences in the toxicity ranking were observed between the conventional culture conditions and the gas-phase models, confirming the importance of correct exposure conditions for the evaluation of toxicity. Both models have advantages that make them suitable as a preliminary step in the toxicity screening of volatile compounds, or for use in a test battery when combined with conventional approaches.

Aldehydes↗

Real time analysis of breath volatiles using SIFT-MS in cigarette smoking.

The selected ion flow tube mass spectrometry (SIFT-MS) technique enables real time analysis of trace volatiles at ppb levels without preconcentration steps or chemical derivatization. Most previous studies of trace compounds on the breath were analyzed using gas chromatography where enhanced detection sensitivity was achieved by concentrating the breath using cryogenic or adsorption trapping techniques. In this paper, we have examined volatile organic substances, isoprene, acetone, ammonia and ethanol in breath before and after smoking a cigarette. It is interesting that isoprene levels increased in all the subjects after smoking one cigarette with a mean increase of 70%. The mean increase for acetone was found to be 22%. In contrast to isoprene, a decreasing ethanol level was observed in all the subjects except one with the negative mean decrease of 28%. Further SIFT-MS studies also have high-lighted some organic substances produced even by unburned cigarettes, US and New Zealand products. Certain US brands have shown much higher levels of volatile species than cigarettes produced in New Zealand.

Acetone↗

Comparison of headspace techniques for sampling volatile natural products in a dynamic system.

Commonly used dynamic sorption techniques for collecting biologically active volatile compounds have been compared. Solid phase microextraction (SPME) using two types of fibers (polydimethylsiloxane, PDMS, 100 microm, and carbowax/divinylbenzene, CW/DVB, 65 microm) was compared to purge and trap methods (Porapak Q, Tenax TA and charcoal) and a technique based on absorption in methanol in a cooling bath. Sampling was done in a stream of purified air (20 ml/min) in a closed and temperature-regulated (27 degrees C) glass tube, passing over a capillary tube containing a hexane solution of tridecane, heptadecane, 1-octen-3-ol, 1-hexadecanol, ethyl tetradecanoate, alpha-pinene, linalool, terpinen-4-ol, cis-verbenol, verbenone, beta-caryophyllene, E,E-farnesol, and geranylgeraniol. With all of the methods, the sampling was performed for a period of 30 min before extraction and analysis was done on a GC-FID system. In general, SPME gave a higher response for all compounds except for alpha-pinene, which was only extracted by the CW/DVB fiber. Purge and trap methods and methanol absorption gave the same response for all substances extracted. None of the methods extracted hexadecanol and geranylgeraniol under the conditions used. However, the SPME equipped with the PDMS coating extracted heptadecane, E,E-farnesol and ethyl tetradecanoate. Our results show that SPME, when selecting the fibers to fit the polarity and volatility of the compounds, is an outstanding extraction method compared to purge and trap and methanol absorption, especially for a qualitative analysis. The best conditions for storing fibers exposed to compounds of high volatility were at low temperatures (6 degrees C) in sealed vials, while the worst way was to leave the exposed fiber unprotected at room temperature (22 degrees C). The dynamic sampling system was effectively tested on a fruiting body of a polypore fungus (Ganoderma applanatum) emitting 1-octen-3-ol, and again SPME showed to be the most sensitive technique.

Biological Products↗