2D spatiotemporal visualization system of expired gaseous ethanol after oral administration for real-time illustrated analysis of alcohol metabolism

A novel 2-dimensional spatiotemporal visualization system of expired gaseous ethanol after oral administration for real-time illustrated analysis of alcohol metabolism has been developed, which employed a low level light CCD camera to detect chemiluminescence (CL) generated by catalytic reactions of standard gaseous ethanol and expired gaseous ethanol after oral administration. First, the optimization of the substrates for visualization and the concentration of luminol solution for CL were investigated. The cotton mesh and 5.0 mmol L⁻¹ luminol solution were selected for further investigations and this system is useful for 0.1-20.0 mmol L⁻¹ of H₂O₂ solution. Then, the effect of pH condition of Tris-HCl buffer solution was also evaluated with CL intensity and under the Tris-HCl buffer solution pH 10.1, a wide calibration range of standard gaseous ethanol (30-400 ppm) was obtained. Finally, expired air of 5 healthy volunteers after oral administration was measured at 15, 30, 45, 60, 75, 90, 105 and 120 min after oral administration, and this system showed a good sensitivity on expired gaseous ethanol for alcohol metabolism. The peaks of expired gaseous ethanol concentration appeared within 30 min after oral administration. During the 30 min after oral administration, the time variation profile based on mean values showed the absorption and distribution function, and the values onward showed the elimination function. The absorption and distribution of expired gaseous ethanol in 5 healthy volunteers following first-order absorption process were faster than the elimination process, which proves efficacious of this system for described alcohol metabolism in healthy volunteers. This system is expected to be used as a non-invasive method to detect VOCs as well as several other drugs [1] in expired air for clinical purpose.     

Continuous-flow analysis for ethanol with immobilized alcohol dehydrogenase

Alcohol dehydrogenase is immobilized by reaction with glutaraldehyde onto silica gel, which is packed in a column in a flow system. Ethanol (0.1–5 mmol) is determined by catalysis of the reaction with NAD⁺, the NADH produced being detected amperometrically. The reproducibility is ca. 1%.     

Non-invasive monitoring of diabetes through analysis of the exhaled breath condensate (aerosol)

We show that exhaled breath condensate (EBC) contains glucose (≈ 0.01 mM for healthy subjects), in contrast to previous works reporting minimal glucose content in EBC. The evaluated breath condensate glucose levels correlate positively with blood glucose levels, thus offering the prospect of a non-invasive approach to the monitoring of diabetes.     

UCSF Chimera--a visualization system for exploratory research and analysis

The design, implementation, and capabilities of an extensible visualization system, UCSF Chimera, are discussed. Chimera is segmented into a core that provides basic services and visualization, and extensions that provide most higher level functionality. This architecture ensures that the extension mechanism satisfies the demands of outside developers who wish to incorporate new features. Two unusual extensions are presented: Multiscale, which adds the ability to visualize large-scale molecular assemblies such as viral coats, and Collaboratory, which allows researchers to share a Chimera session interactively despite being at separate locales. Other extensions include Multalign Viewer, for showing multiple sequence alignments and associated structures; ViewDock, for screening docked ligand orientations; Movie, for replaying molecular dynamics trajectories; and Volume Viewer, for display and analysis of volumetric data. A discussion of the usage of Chimera in real-world situations is given, along with anticipated future directions. Chimera includes full user documentation, is free to academic and nonprofit users, and is available for Microsoft Windows, Linux, Apple Mac OS X, SGI IRIX, and HP Tru64 Unix from http://www.cgl.ucsf.edu/chimera/.     

Improved real-time puff-by-puff GC-MS system for whole smoke analysis

An automated puff-by-puff mainstream smoke (MSS) system is developed to monitor real-time whole smoke in mainstream cigarette smoke using gas chromatography (GC)-mass spectrometry (MS). The whole-smoke analysis is based on automated sample collection and injection into the GC-MS system. The important feature of this system is the real-time rapid analysis that is simple, sensitive, precise, flexible, and exhibits low carryover of volatile and semivolatile smoke constituents. The system is equipped with an automated sampling and switching valve and a smoking machine. The key improvements of the system, as compared with current and alternative methodologies, include minimizing variations caused by operator sampling techniques, the real-time analysis of MSS, the detection of flavorants in MSS from a single puff of cigarette smoke, the ability to analyze numerous smoke constituents from either whole smoke or the gas phase of a single puff, the ability to monitor a few selected smoke constituents in whole smoke using multiple puffs, and its good feasibility compared with solvent extraction and impinger trapping procedures for volatile organic compounds in MSS. System configuration and sampling methodologies are described. Sensitivity, flexibility, precision, feasibility, carryover, and applications of the system are discussed.     

2-(Trimethylsilyl)ethanol as a new alcohol equivalent for copper-catalyzed coupling of aryl iodides

2-(Trimethylsilyl)ethanol as a new alcohol equivalent has been employed for copper-catalyzed coupling of aryl iodides. Using mild reaction conditions, it has been observed that substituted phenols and phenols with sensitive functional groups can be readily prepared.     

Enzymatic microreactors for the determination of ethanol by an automatic sequential injection analysis system

A sequential injection analysis system with two enzymatic microreactors for the determination of ethanol has been designed. Alcohol oxidase and horseradish peroxidase were separately immobilized on glass aminopropyl beads, and packed in 0.91-mL volume microreactors, working in line with the sequential injection analysis system. A stop flow of 120 s was selected for a linear ethanol range of 0.005-0.04 g/L +/- 0.6% relative standard deviation with a throughput of seven analyses per hour. The system was applied to measure ethanol concentrations in samples of distilled and nondistilled alcoholic beverages, and of alcoholic fermentation with good performance and no significant difference compared with other analytical procedures (gas chromatography and high-performance liquid chromatography).     

An investigation into the electro-oxidation of ethanol and 2-propanol for application in direct alcohol fuel cells (DAFCs)

A comparative study of the electro-oxidation of ethanol and 2-propanol was carried out on carbon-supported platinum particles. Cyclic voltammetry, steady state polarisation, and electrochemical impedance spectroscopy were used to investigate the oxidation reactions. A difference in the mechanistic behaviour of the oxidation of ethanol and 2-propanol on Pt was observed, thereby highlighting the fact that the molecular structure of the alcohol has great influence on its electroreactivity. The study emphasizes the fact that 2-propanol is a promising fuel candidate for a direct alcohol fuel cell.     

NaNO2-activated, iron-TEMPO catalyst system for aerobic alcohol oxidation under mild conditions

FeCl3-TEMPO-NaNO2 catalyses the selective and mild aerobic oxidation of a broad range of alcohols to the corresponding aldehydes and ketones.     

A sensitive chemiluminescence method for the determination of H2O2 in exhaled breath condensate

In this paper, a novel flow injection chemiluminescence (FI-CL) method is proposed for the determination of picomolar L(-1) levels of hydrogen peroxide (H(2)O(2)) in exhaled breath condensate (EBC). This method is based on the oxidation of a low concentration of luminol (10(-7) M) by H(2)O(2) at a low concentration level (< 10(-8) M) in an alkaline medium catalyzed by a complex, K(5)[Cu(HIO(6))(2)] (DPC), which is not interfered by other metal ions or horseradish peroxidase (HRP). Under the optimum conditions, H(2)O(2) was determined over the range of 1.0 x 10(-10) to 1.0 x 10(-8) mol L(-1) with a detection limit of (3sigma) of 4.1 x 10(-11) mol L(-1). The relative standard deviation (RSD) was 3.2% for 5 nmol L(-1) H(2)O(2) (n = 7). The proposed method offers the advantages of ultra-sensitivity, selectivity, simplicity and rapidity for H(2)O(2) determination. It was successfully applied to directly determine trace amounts of H(2)O(2) (nmol L(-1)) in human's EBC of both rheum and healthy volunteers. A statistically significant difference was found between patients with rheum (n = 11) and control subjects without rheum (n = 11).     

Kinetics of ethanol decay in mouth‐ and nose‐exhaled breath measured on‐line by selected ion flow tube mass spectrometry following varying doses of alcohol

A study has been carried out of the decay of ethanol in mouth‐exhaled and nose‐exhaled breath of two healthy volunteers following the ingestion of various doses of alcohol at different dilutions in water. Concurrent analyses of sequential single breath exhalations from the two volunteers were carried out using selected ion flow tube mass spectrometry, SIFT‐MS, on‐line and in real time continuously over some 200 min following each alcohol dose by simply switching sampling between the two volunteers. Thus, the time interval between breath exhalations was only a few minutes, and this results in well‐defined decay curves. Inspection of the mouth‐exhaled and nose‐exhaled breath data shows that mouth contamination of ethanol diminished to insignificant levels after a few minutes. The detailed results of the analyses of nose‐exhaled breath show that the peak levels and the decay rates of breath ethanol are dependent on the ethanol dose and the volume of ethanol/water mixture ingested. From these data, both the efficiency of the first‐pass metabolism of ethanol and the indications of gastric emptying rates at the various doses and ingested volumes have been obtained for the two volunteers. Additionally and simultaneously, acetaldehyde, acetic acid and acetone were measured in each single breath exhalation. Acetaldehyde, the primary product of ethanol metabolism, is seen to track the breath ethanol. Acetic acid, a possible secondary product of this metabolism, was detected in the exhaled breath, but was shown to largely originate in the oral cavity. Breath acetone was seen to increase over the long period of measurement due to the depletion of nutrients. Copyright © 2010 John Wiley & Sons, Ltd.     

Toluidine blue adsorbed on alcohol dehydrogenase modified glassy carbon electrode for voltammetric determination of ethanol

A novel toluidine blue O (TBO) adsorbed alcohol dehydrogenase (ADH) biocomposite film have been prepared through simple adsorption technique with the help of electrostatic interaction between oppositely charged layers. Nafion (NF) coating was made on top of the biocomposite film modified glassy carbon electrode (GCE) to protect ADH from leaching. The fabricated ADH/TBO/NF biocomposite electrode remains highly stable in the pH range from 4 to 13. More facile electron transfer process occurs at ADH/TBO/NF biocomposite than at TBO/NF film, which is obvious from the six folds increase in k_s value. Maximum surface coverage concentration (Γ) of TBO is noticed at ADH/TBO/NF film, which is 82% higher than at TBO/NF and 15% higher than at ADH/TBO film modified GCEs. Electrochemical impedance spectroscopy studies reveal that ADH has been well immobilized in the biocomposite film. Scanning electron microscopy studies confirm the discriminate surface morphology of various components present in the biocomposite film. Cyclic voltammetry studies validate that ADH/TBO/NF biocomposite film exhibits excellent electrocatalytic activity for ethanol oxidation at low over potential (I_pa = −0.14 V). The same studies show biocomposite film possesses a good sensitivity of 7.91 μA M⁻¹ cm⁻² for ethanol determination. This above sensitivity value is 17.40% higher than the sensitivity obtained for TBO/NF film (6.74 μA M⁻¹ cm⁻²). Further, using differential pulse voltammetry, a sensitivity of 1.70 μA M⁻¹ cm⁻² has been achieved for ADH/TBO/NF biocomposite film.     

Amperometric sensor for ethanol based on one-step electropolymerization of thionine-carbon nanofiber nanocomposite containing alcohol oxidase

Thionine had strong interaction with carbon nanofiber (CNF) and was used in the non-covalent functionalization of carbon nanofiber for the preparation of stable thionine-CNF nanocomposite with good dispersion. With a simple one-step electrochemical polymerization of thionine-CNF nanocomposite and alcohol oxidase (AOD), a stable poly(thionine)-CNF/AOD biocomposite film was formed on electrode surface. Based on the excellent catalytic activity of the biocomposite film toward reduction of dissolved oxygen, a sensitive ethanol biosensor was proposed. The ethanol biosensor could monitor ethanol ranging from 2.0 to 252 μM with a detection limit of 1.7 μM. It displayed a rapid response, an expanded linear response range as well as excellent reproducibility and stability. The combination of catalytic activity of CNF and the promising feature of the biocomposite with one-step non-manual technique favored the sensitive determination of ethanol with improved analytical capabilities.     

Novel hierarchical classification and visualization method for multiobjective optimization of drug properties: application to structure-activity relationship analysis of cytochrome P450 metabolism

In the lead optimization process, medicinal chemists must consider various chemical properties of active compounds, including ADME/Tox properties, and find the best compromise among these. This study presents a novel data mining method for multiobjective optimization of chemical properties, which consists of the hierarchical classification and visualization of multidimensional data. A hierarchical classification tree model is generated by an extension of recursive partitioning that utilizes averaged information gains for multiple objective variables as a quality-of-split criterion. All the hierarchically structured data objects are represented using a large-scale data visualization technique. The technique is an extension of HeiankyoView, which displays data objects as colored icons and group nodes as rectangular borders. Each icon is divided into subregions with different colors, so that it can present multidimensional data according to brightness of the colors. The proposed method was applied to the structure-activity relationship analysis for cytochrome P450 (CYP) substrates. The substrate specificity of six CYP isoforms was successfully delineated: e.g., CYP2C9 substrates are anionic compounds, while CYP2D6 substrates are cationic; and CYP2E1 substrates are smaller compounds, while CYP3A4 substrates are larger compounds.     

Evaluation of CD detection in an HPLC system for analysis of DHEA and related steroids

The biological importance of dehydroepiandrosterone (DHEA) is reflected by the fact that DHEA is a crucial precursor of the biosynthesis of the steroidal sex hormones. Simultaneous separation of DHEA, dehydroepiandrosterone sulfate (DHEA-S), pregnenolone, androstenedione and testosterone has been accomplished by reversed-phase ion-pair high-performance liquid chromatography (RP-IP-HPLC) based on isocratic elution applying circular dichroism (CD) detection at 295 nm. Addition of tetrabutylammonium hydrogensulfate to the mobile phase increases the retention of DHEA-S on the C8-silica column by an apparent ion-pairing mechanism without affecting the retention of the other (non-ionic) steroids. CD spectroscopy provides highly selective detection of compounds possessing optically active absorption bands and the separation is even more selective in the higher wavelength range applied. The linearity of the steroid concentration (c, mg mL⁻¹) versus peak area was tested in the concentration range of 0.5–2 mg mL⁻¹ (injected quantities were 10–40 μg). The relative standard deviation (RSD) values for DHEA and DHEA-S indicated a good intra-assay and inter-assay precision of the method.      

Quantum theory of atoms in molecules analysis on the conformational preferences of vinyl alcohol and related ethers

Vinyl alcohol, methyl vinyl ether, and tert-butyl vinyl ether were studied within the framework of the quantum theory of atoms in molecules at the B3LYP/6-311++G(2d,2p) level. Local and integrated properties of the charge density indicate that the anti conformational preference of the tert-butyl derivative is not due to a differing resonance contribution with regard to the less bulky vinyl ethers but to steric effects. There is a small delocalization of charge density, either total or pi, between oxygen and the terminal vinyl carbon, which does not support the resonance picture of vinyl compounds.     

A turbidimetric micromethod for analysis of tertiary butyl alcohol

Summary A rapid turbidimetric method for the analysis of very dilute aqueous solutions of tertiary butyl alcohol has been developed, using a mercuric sulphate/sulphuric acid reagent similar to the Denigés reagent. Concentrations as low as 1–6 mg/litre can be determined to within about 5% error.

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Zusammenfassung Eine schnelle turbidimetrische Methode zur Bestimmung sehr verdünnter, wäßriger Lösungen von tertiärem Butylalkohol wurde beschrieben. Sie beruht auf der Reaktion mit einer dem Reagens vonDenigés ähnlichen Quecksilber(II)-sulfat-Schwefelsäure-Lösung. Konzentrationen von 1 bis 6 mg/1 können mit einer Genauigkeit von etwa 5% bestimmt werden.

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Résumé L'auteur a mis au point une méthode turbidimétrique rapide pour l'analyse de solutions aqueuses très diluées d'alcool butylique tertiaire; à cette fin il emploie un réactif acide sulfurique-sulfate mercurique semblable à celui deDenigès. Dans ces conditions il est possible de déterminer des concentrations s'abaissant à 1–6 mg/litre avec une erreur inférieure à environ 5%.

     

Dihydroisoxazole analogs for labeling and visualization of catalytically active transglutaminase 2

We report the synthesis and preliminary characterization of "clickable" inhibitors of human transglutaminase 2 (TG2). These inhibitors possess the 3-halo-4,5-dihydroisoxazole warhead along with bioorthogonal groups such as azide or alkyne moieties that enable subsequent covalent modification with fluorophores. Their mechanism for inhibition of TG2 is based on halide displacement, resulting in the formation of a stable imino thioether. Inhibition assays against recombinant human TG2 revealed that some of the clickable inhibitors prepared in this study have comparable specificity as benchmark dihydroisoxazole inhibitors reported earlier. At low micromolar concentrations they completely inhibited transiently activated TG2 in a WI-38 fibroblast scratch assay and could subsequently be used to visualize the active enzyme in?situ. The potential use of these inhibitors to probe the role of TG2 in celiac sprue as well as other diseases is discussed.