Anticonflict effects of rose oil and identification of its active constituents

The present study investigates the pharmacologically active constituents of rose oil, which possesses anti-conflict effects. Analysis using GC/MS revealed that rose oil contains 9 substances that were identified as myrcene, benzyl alcohol, 2-phenethyl alcohol, citronellol, geraniol, citronellyl acetate, eugenol, geranyl acetate and methyl eugenol. We examined the effects of each of these substances using the Geller and Vogel conflict tests in ICR mice. Myrcene, benzyl alcohol and citronellyl acetate did not produce any effects in either tests. Geranyl acetate and methyl eugenol produced no effect in the Geller conflict test. Geraniol and eugenol decreased the response rate during the safe period of the Geller conflict test, but did not affect the response rate during the alarm period. In contrast, 2-phenethyl alcohol and citronellol, like rose oil, produced an increasing effect on the response rate during the alarm period in the Geller conflict test. In addition, both chemicals increased the number of electric shocks mice received in the Vogel conflict test in a manner similar to that of rose oil. Given that 2-phenethyl alcohol and citronellol produced the same anti-conflict effects in both tests as rose oil, we concluded that they are the pharmacologically active constituents of anti-anxiety-like effect of rose oil.     

Coupling of Metabolism and Bioconversion: Microbial Esterification of Citronellol with Acetyl Coenzyme A Produced via Metabolism of Glucose in an Interface Bioreactor

Microbial esterification of citronellol with acetyl coenzyme A (acetyl-CoA) produced via metabolism of glucose was performed with Hansenula saturnus IFO 0809 in an interface bioreactor by using a hydrophilic carrier (an agar plate or an agar-coated filter pad) and a hydrophobic organic solvent (decane). An increase in the glucose concentration on an agar plate led to a decrease in oxidation of citronellol and an increase in the amount of citronellyl acetate. Fed-batch addition of citronellol efficiently alleviated substrate toxicity and resulted in the accumulation of high levels of citronellyl acetate. Furthermore, triple coupling of acetyl-CoA formation, microbial reduction of citronellal to citronellol, and esterification of the resulting citronellol with acetyl-CoA was also an efficient way to prepare citronellyl acetate. By using coupling systems, citronellyl acetate could be efficiently produced without any acetyl donor.     

Lipase catalysed synthesis of aliphatic, terpene and aromatic esters by alcoholysis in solvent-free medium

Biosynthesis of pentyl and terpene valerates, terpene acetates, hexyl and benzyl benzoates was conducted via alcoholysis, at 55 °C, in a solvent-free medium, using lipases from Candida antarcticaand Mucor mieheias catalysts. The yield higher than 90% was achieved (Candida antarctica)after 10 h for synthesis of pentyl valerate and 3-methylbutyl valerate, 35 h for 2-methylbytyl valerate, 6 h for geranyl and cytronelyl acetates, 10 h for geranyl and cytronelyl valerates, 75 h for hexyl benzoate and 100 h for benzyl benzoate.     

Evaluation of different approaches for lipase catalysed synthesis of citronellyl acetate

Three different approaches for lipase catalysed synthesis of citronellyl acetate by a commercial available immobilized lipase have been studied: a) direct esterification reaction of citronellol with acetic acid; b) alcoholysis of butyl acetate with citronellol and c) transesterification of citronellyl butyrate with butylacetate. Heptane was used as solvent for all the experiments. The extent of reaction occurred in the order alcoholysis>transesterification>esterification. Substrate partitioning between the immobilization support and the organic medium seems to greatly influence the catalytic performance of this enzyme preparation. Production of citronellyl acetate, was found to be dependent on the partition coefficient of the acyl donor which account the greatest value for the acetic acid.     

Some Neutral Components of Cigarette Smoke (I)

The neutral fraction of cigarette smoke Components, produced in constant-volume continuous smoking by an artificial smoking device, was sub-fractionated into alcohols, acid-, alcohol- and phenol-components of esters, unsaponifiable matters and hydrocarbons. From these sub-fractions ethyl alcohol, benzyl alcohol, β-phenylethyl alcohol and ethyl acetate were first isolated as their crystalline derivatives and then identified by their melting points, infrared spectra and elementary analyses. It is worth to note that the presence of these compounds in tobacco smoke has not yet been reported in the literature.     

Inhibition by cigarette smoke of lipid peroxidation-induced neurotransmitter release

Effects of water-soluble substance in cigarette smoke on neurotransmitter release were investigated using nerve terminals (synaptosomes) prepared from rat cerebral cortex. 2,2′-Azobis (2-amidinopropane) dihydrochloride (ABAP), a peroxyl radical-generator, enhanced the depolarization-evoked release of glutamate and aspartate from synaptosomes with concomitant increase in thiobarbituric acid-reactive substances (TBA-RS) levels in membrane lipids of synaptosomes. The trapped smoke-substance attenuated the lipid peroxidation-enhanced release of excitatory amino acids during the depolarization with reduction in TBA-RS, although it failed to affect the basal release of neurotransmitters. These data suggest that cigarette smoke may possess antioxidant properties to reduce oxidation-induced enhancement of transmitter release from nerve terminals.     

Exposure to Biomass Smoke Extract Enhances Fibronectin Release from Fibroblasts

COPD induced following biomass smoke exposure has been reported to be associated with a more fibrotic phenotype than cigarette smoke induced COPD. This study aimed to investigate if biomass smoke induced extracellular matrix (ECM) protein production from primary human lung fibroblasts in vitro. Primary human lung fibroblasts (n = 5–10) were stimulated in vitro for up to 72 hours with increasing concentrations of biomass smoke extract (BME) or cigarette smoke extract (CSE) prior to being assessed for deposition of ECM proteins, cytokine release, and activation of intracellular signalling molecules. Deposition of the ECM proteins perlecan and fibronectin was upregulated by both CSE (p<0.05) and BME (p<0.05). The release of the neutrophilic chemokine IL-8 was also enhanced by BME. ERK1/2 phosphorylation was significantly upregulated by BME (p<0.05). Chemical inhibition of ERK signalling molecules partially attenuated these effects (p<0.05). Stimulation with endotoxin had no effect. This study demonstrated that BME had similar effects to CSE in vitro and had the capacity to directly induce fibrosis by upregulating production of ECM proteins. The mechanisms by which both biomass and cigarette smoke exposure cause lung damage may be similar.     

Mouse Protocadherin-1 Gene Expression Is Regulated by Cigarette Smoke Exposure In Vivo

Protocadherin-1 (PCDH1) is a novel susceptibility gene for airway hyperresponsiveness, first identified in families exposed to cigarette smoke and is expressed in bronchial epithelial cells. Here, we asked how mouse Pcdh1 expression is regulated in lung structural cells in vivo under physiological conditions, and in both short-term cigarette smoke exposure models characterized by airway inflammation and hyperresponsiveness and chronic cigarette smoke exposure models. Pcdh1 gene-structure was investigated by Rapid Amplification of cDNA Ends. Pcdh1 mRNA and protein expression was investigated by qRT-PCR, western blotting using isoform-specific antibodies. We observed 87% conservation of the Pcdh1 nucleotide sequence, and 96% conservation of the Pcdh1 protein sequence between men and mice. We identified a novel Pcdh1 isoform encoding only the intracellular signalling motifs. Cigarette smoke exposure for 4 consecutive days markedly reduced Pcdh1 mRNA expression in lung tissue (3 to 4-fold), while neutrophilia and airway hyperresponsiveness was induced. Moreover, Pcdh1 mRNA expression in lung tissue was reduced already 6 hours after an acute cigarette-smoke exposure in mice. Chronic exposure to cigarette smoke induced loss of Pcdh1 protein in lung tissue after 2 months, while Pcdh1 protein levels were no longer reduced after 9 months of cigarette smoke exposure. We conclude that Pcdh1 is highly homologous to human PCDH1, encodes two transmembrane proteins and one intracellular protein, and is regulated by cigarette smoke exposure in vivo.     

Volatile constituents of Trichothecium roseum

In the course of investigation of Trichothecium roseum (Fungi Imperfecti) for its attractancy against Tyrophagus putrescentiae (cheese mite), the twenty following volatile compounds produced at a very low concentration by the microfungus were identified by gc, gc/ms, gc/c.i.ms and tlc: 3-methyl-1-butanol, 3-octanone, 1-octen-3-one, 3-octanol, octa-1,5-dien-3 one, 1-octen-3-ol, 6-methyl-5-hepten-2-ol, octa-1,5-dien-3 ol, furfural, linalool, linalyl acetate, terpineol (alpha and beta) citronellyl acetate, nerol, citronellol, phenylacetaldehyde, benzyl alcohol geranyl acetate, 1-phenyl ethanol and nerolidol. Octa-1,5-dien-3-ol and octa-1,5-dien-3-one have not been previously isolated from fungi; octa-1,5-dien-3-ol is the most potent attractant amount the volatile compounds detected by gc.     

Quantitative Studies on Fabrics as Disseminators of Viruses: III. Persistence of Vaccinia Virus on Fabrics Impregnated with a Virucidal Agent

Eight compounds were tested in vitro for virucidal and antiviral activity against poliovirus and vaccinia virus. These compounds included five quaternary ammonium salts, two bromosalicylanilides, and neomycin sulfate, an antibiotic. None of the compounds was active against poliovirus, but virucidal activity was demonstrated against vaccinia virus with three of the quarternary ammonium compounds: n-alkyl (C14, C12, C16) dimethyl benzyl ammonium chloride, di-isobutyl cresoxy ethoxy ethyl dimethyl benzyl ammonium chloride monohydrate, and n-alkyl (60% C14, 30% C16, 5% C12, 5% C18) dimethyl benzyl ammonium chlorides plus n-alkyl (50% C12, 30% C14, 17% C16, 3% C18) dimethyl ethylbenzyl ammonium chlorides. Wool blanketing, wool gabardine, and cotton sheeting materials were impregnated with the first of the above virucidal compounds, and the persistence of vaccinia virus on these fabrics was compared with the persistence of the agent on nonimpregnated fabrics of the same type held at 25 C in 35 and 78% relative humidity. No virus could be recovered from the chemically treated fabrics at any time after virus exposure, whereas the virus persisted as long as 4 weeks on nonimpregnated materials. Viable vaccinia virus was also found to persist less than 1 day on a cotton fabric finished with a wash-and-wear modified triazone resin. Poliovirus persisted less than 5 days on this wash-and-wear fabric.     

Amino Acid Regions of Family 45 Endoglucanases Involved in Cotton Defibrillation and in Resistance to Anionic Surfactants and Oxidizing Agents

In the detergent industry, fungal endoglucanases are used to release microfibrils from the surfaces of dyed cellulosic fabrics to enhance color brightness. Family 45 endoglucanase (glycoside hydrolase family 45, GH45) EGL3 from Humicola grisea is more resistant to anionic surfactants and oxidizing agents than family 45 endoglucanase RCE1 from Rhizopus oryzae, while in the present study, a catalytic domain of RCE1 had higher defibrillation activity on dyed cotton fabrics than did that of EGL3. To identify the amino acid regions involved in these properties, we compared the characteristics of RCE1, EGL3, and three chimeric endoglucanases, in which each of the three regions of the catalytic domain of EGL3 was replaced by the corresponding region of the catalytic domain of RCE1. Amino acids in the N-terminal region were involved in resistance to anionic surfactants and oxidizing agents. Furthermore, amino acids in the region adjacent to the N-terminal region were involved in releasing microfibrils and in binding to dyed cotton fabrics, indicating that the binding of the amino acids in this region might be important in the release of microfibrils from dyed cotton fabrics.     

香叶油的化学成分

为了探讨不同产地香叶油的香气特徵,我们对云南省昆明、玉溪、石屏、宾川产香叶油和法国波尔蓬,埃及产香叶油的化学成分用Finnigan-4510型气相色谱/质谱/计算机联用方法进行了定性、定量分析,共分离鉴定了35个成分(见表2)。 分析结果表明:云南产香叶油的香气成分主要由香茅醇,甲酸香茅酯,香叶醇,玫瑰醚构成;而法国、埃及产香叶油的香气成分则主要由香叶醇,甲酸香叶酯,香茅醇,甲酸香茅酯和芳樟醇构成。二者具有不同的格调和香气特徵。     

Composition of turpentine from Pinus edulis wood oleoresin

Monoterpenoids and sesquiterpenoid hydrocarbons of Pinus edulis wood oleoresin were analyzed by chromatographic and spectroscopic methods. Monoterpenoid hydrocarbons (20·3%) were composed mainly of α-pinene, with camphene, β-pinene, 3-carene, sabinene, myrcene, limonene, β-phellandrene, trans-ocimene and terpinolene in secondary to trace amounts. Oxygenated terpenoids (0.28%) contained bornyl acetate and verbenone as major constituents, and linalool, camphor, terpinene-4-ol, citronellyl acetate, borneol, neral, α-terpineol, citronellol, nerol, and geraniol in smaller amounts. Oleoresin contained 1·1% of acetogenins, composed mainly of ethyl caprylate. Sesquiterpenoid hydrocarbons were high (5·7%) in oleoresin) and were composed of germacrene D as a major constituent (36·6%), of γ-amorphene, α-copaene, and longifolene as secondary constituents (5–20%), and β-farnesene, α- and γ-murolenes, β₁-, γ-, δ-, and ε-cadinenes, α-amorphene, δ-guaiene, sibirene, α-cubebene, β-copaene, β-ylangene, sativene, cyclosativene, β-bourbonene, α- and γ-humulenes, caryophyllene, α-longipinene and longicyclene in smaller amounts. Composition of P. edulis and of P. monophylla turpentines was found to be similar, with percentage of ethyl caprylate being the best distinguishing criterion.     

Chemical composition of volatiles from cortical oleoresin of Pseudotsuga menziesii

Pseudotsuga menziesii cortical oleoresin was found to contain 1·7% of oxygenated terpenoids and compounds of similar volatility composed of linalool, methylsalicylate, bornyl acetate, citronellol, geranyl acetate, methylthymol, citronellyl acetate, terpinen-4-ol, borneol, isopulegol, anethole, terpinen-4-ol acetate, camphor, geraniol, neryl acetate, and nerol. Sesquiterpenoid hydrocarbons were low (only 0·07%) and contained sibirene and longifolene as main constituents, with β-caryophyllene, γ-muurolene, γ-cadinene (identified by IR), and 20 additional compounds in small amounts. p-Cymen-8-ene was identified in monoterpene hydrocarbon fraction.     

Amino acid regions of family 45 endoglucanases involved in cotton defibrillation and in resistance to anionic surfactants and oxidizing agents

In the detergent industry, fungal endoglucanases are used to release microfibrils from the surfaces of dyed cellulosic fabrics to enhance color brightness. Family 45 endoglucanase (glycoside hydrolase family 45, GH45) EGL3 from Humicola grisea is more resistant to anionic surfactants and oxidizing agents than family 45 endoglucanase RCE1 from Rhizopus oryzae, while in the present study, a catalytic domain of RCE1 had higher defibrillation activity on dyed cotton fabrics than did that of EGL3. To identify the amino acid regions involved in these properties, we compared the characteristics of RCE1, EGL3, and three chimeric endoglucanases, in which each of the three regions of the catalytic domain of EGL3 was replaced by the corresponding region of the catalytic domain of RCE1. Amino acids in the N-terminal region were involved in resistance to anionic surfactants and oxidizing agents. Furthermore, amino acids in the region adjacent to the N-terminal region were involved in releasing microfibrils and in binding to dyed cotton fabrics, indicating that the binding of the amino acids in this region might be important in the release of microfibrils from dyed cotton fabrics.     

Insights into structural features determining odorant affinities to honey bee odorant binding protein 14

Molecular interactions between odorants and odorant binding proteins (OBPs) are of major importance for understanding the principles of selectivity of OBPs towards the wide range of semiochemicals. It is largely unknown on a structural basis, how an OBP binds and discriminates between odorant molecules. Here we examine this aspect in greater detail by comparing the C-minus OBP14 of the honey bee (Apis mellifera L.) to a mutant form of the protein that comprises the third disulfide bond lacking in C-minus OBPs. Affinities of structurally analogous odorants featuring an aromatic phenol group with different side chains were assessed based on changes of the thermal stability of the protein upon odorant binding monitored by circular dichroism spectroscopy. Our results indicate a tendency that odorants show higher affinity to the wild-type OBP suggesting that the introduced rigidity in the mutant protein has a negative effect on odorant binding. Furthermore, we show that OBP14 stability is very sensitive to the position and type of functional groups in the odorant.     

How Far Does a Receptor Influence Vibrational Properties of an Odorant?

The biophysical mechanism of the sense of smell, or olfaction, is still highly debated. The mainstream explanation argues for a shape-based recognition of odorant molecules by olfactory receptors, while recent investigations suggest the primary olfactory event to be triggered by a vibrationally-assisted electron transfer reaction. We consider this controversy by studying the influence of a receptor on the vibrational properties of an odorant in atomistic details as the coupling between electronic degrees of freedom of the receptor and the vibrations of the odorant is the key parameter of the vibrationally-assisted electron transfer. Through molecular dynamics simulations we elucidate the binding specificity of a receptor towards acetophenone odorant. The vibrational properties of acetophenone inside the receptor are then studied by the polarizable embedding density functional theory approach, allowing to quantify protein-odorant interactions. Finally, we judge whether the effects of the protein provide any indications towards the existing theories of olfaction.     

MiR-101 and miR-144 Regulate the Expression of the CFTR Chloride Channel in the Lung

The Cystic Fibrosis Transmembrane conductance Regulator (CFTR) is a chloride channel that plays a critical role in the lung by maintaining fluid homeostasis. Absence or malfunction of CFTR leads to Cystic Fibrosis, a disease characterized by chronic infection and inflammation. We recently reported that air pollutants such as cigarette smoke and cadmium negatively regulate the expression of CFTR by affecting several steps in the biogenesis of CFTR protein. MicroRNAs (miRNAs) have recently received a great deal of attention as both biomarkers and therapeutics due to their ability to regulate multiple genes. Here, we show that cigarette smoke and cadmium up-regulate the expression of two miRNAs (miR-101 and miR-144) that are predicted to target CFTR in human bronchial epithelial cells. When premature miR-101 and miR-144 were transfected in human airway epithelial cells, they directly targeted the CFTR 3′UTR and suppressed the expression of the CFTR protein. Since miR-101 was highly up-regulated by cigarette smoke in vitro, we investigated whether such increase also occurred in vivo. Mice exposed to cigarette smoke for 4 weeks demonstrated an up-regulation of miR-101 and suppression of CFTR protein in their lungs. Finally, we show that miR-101 is highly expressed in lung samples from patients with severe chronic obstructive pulmonary disease (COPD) when compared to control patients. Taken together, these results suggest that chronic cigarette smoking up-regulates miR-101 and that this miRNA could contribute to suppression of CFTR in the lungs of COPD patients.     

Operating Mechanism and Molecular Dynamics of Pheromone-Binding Protein ASP1 as Influenced by pH

Odorant binding protein (OBP) is a vital component of the olfactory sensation system. It performs the specific role of ferrying odorant molecules to odorant receptors. OBP helps insects and types of animal to sense and transport stimuli molecules. However, the molecular details about how OBPs bind or release its odorant ligands are unclear. For some OBPs, the systems'' pH level is reported to impact on the ligands'' binding or unbinding capability. In this work we investigated the operating mechanism and molecular dynamics in bee antennal pheromone-binding protein ASP1 under varying pH conditions. We found that conformational flexibility is the key factor for regulating the interaction of ASP1 and its ligands, and the odorant binds to ASP1 at low pH conditions. Dynamics, once triggered by pH changes, play the key roles in coupling the global conformational changes with the odorant release. In ASP1, the C-terminus, the N-terminus, helix α2 and the region ranging from helices α4 to α5 form a cavity with a novel ‘entrance’ of binding. These are the major regions that respond to pH change and regulate the ligand release. Clearly there are processes of dynamics and hydrogen bond network propagation in ASP1 in response to pH stimuli. These findings lead to an understanding of the mechanism and dynamics of odorant-OBP interaction in OBP, and will benefit chemsensory-related biotech and agriculture research and development.