Contribution of uniformly 13C-enriched sterigmatocystin to the study of its pulmonary metabolism

Mycotoxins are secondary metabolites of filamentous fungi which can cause a wide range of systemic effects. Human health effects of inhaled mycotoxins remain poorly documented, despite the large amounts present, associated with air-borne particles. Among these mycotoxins, sterigmatocystin is one of the most prevalent. Because its chemical structure is close to that of the aflatoxins, we studied its metabolism and its cellular consequences when in contact with the airway epithelium, using the mass spectral signature from the 10% (13)C uniformly enriched sterigmatocystin. The metabolism was studied in vitro, using recombinant cytochrome P450s enzymes, and in porcine tracheal epithelial cell (PTEC) primary cultures at an air-liquid interface. The metabolites were analyzed by high-performance liquid chromatography coupled with tandem mass spectrometry detection. Expressed enzymes and PTECs were exposed to uniformly (13)C-enriched sterigmatocystin to confirm the relationship between sterigmatocystin and its metabolites because this isotopic cluster shape is conserved for all metabolites and their product ions. Incubation of sterigmatocystin with recombinant cytochrome P450 1A1 led to the formation of three metabolites identified as monohydroxysterigmatocystin, dihydroxysterigmatocystin and one glutathione adduct, the latter after the formation of a transient intermediate. In the PTEC cultures, sterigmatocystin metabolism resulted in a glucuro-conjugate. Two other products were detected, a sulfo-conjugate and a glucuro-conjugate of hydroxysterigmatocystin upon cytochrome P450 1A1 induction. This is the first study to report sterigmatocystin metabolism in airway epithelium, and it suggests that, contrary to the aflatoxins, sterigmatocystin is mainly detoxified into its conjugates and is unable to produce significant amounts of reactive metabolites in respiratory cells, at least in pigs.     

The production and characterisation of an antibody to detect the coccidiostat toltrazuril and its metabolite ponazuril

The production of an antibody to detect toltrazuril or its metabolite ponazuril is complicated due to structural constraints of conjugating these coccidiostats to a carrier protein. Therefore a search was carried out for a compound that shared a common substructure to use as an antigen mimic. The chosen compound, trifluoraminoether, was conjugated to two carrier proteins (HSA and BTG) and used in the immunisation of six rabbits. Two immunogen doses (1 mg and 0.1 mg) were also used. All six rabbits produced an immunological response to the hapten regardless of the carrier protein or immunogen dose used. The most sensitive polyclonal antibody produced, designated R609, was subsequently characterised. This antiserum exhibited an IC50 of 18 ng ml(-1) using a competitive ELISA format. Cross reactivity studies show that this serum is specific for toltrazuril and its metabolites (toltrazuril sulfoxide and toltrazuril sulfone) but does not cross-react with other coccidiostats such as halofuginone, nitroimidazoles or nicarbazin. This is the first reported production of an antibody capable of specifically binding toltrazuril and ponazuril.     

Microwave specific Wolff rearrangement of alpha-diazoketones and its relevance to the nonthermal and thermal effect

alpha-Diazoketones possess high electric dipole moments, as a consequence of the dipolar nature of the diazocarbonyl functional group. The vectorial analysis, theoretical calculations (PM3 and ab initio), and literature reports based on experimental and theoretical calculations reveal a higher dipole moment for the Z-configuration of the diazo functional group. Microwave irradiation of alpha-diazoketone (1a-m) (Figure 1) promotes Wolff rearrangement specifically via the Z-configuration in excellent yields. The dielectric properties of the solvent govern the course of the microwave rearrangement. 3-Diazocamphor (1m) on microwave irradiation in benzylamine exhibits nonthermal effects to furnish exclusively the Wolff rearrangement product (4m), equivalent to its photochemical behavior. In the presence of an aqueous medium, through solvent heating predominates, leading to the formation of a tricyclic ketone (5) as the principal product, arising from an intramolecular C-H insertion. This behavior is similar to its known thermal and transition metal catalyzed reactivity pattern.     

Variable-pitch dispensing workstation and its application to the preparation of microsensor arrays.

We have developed an 8-ch capillary-based dispensing workstation with a variable capillary pitch mechanism. The capillary intervals can be varied from 1 to 9 mm to dispense different solutions simultaneously at an arbitrary dispensing pitch, allowing direct dispensing from microplates to integrated analytical systems. To evaluate the precision of its dispensing performance, droplets of Rhodamine G dye were dispensed onto glass slides and the values of the optical volume were analyzed. The error in the dispensed volume proved to be 0.54 nL when dispensing 20 nL. In dispensing small volumes, the volume error for this workstation was found to be about 100-fold less than that seen in conventional dispensers. Even highly viscous solutions containing 50% glycerol could be dispensed with precision. Rapid dispensing was also achieved. Moreover, the application of the workstation to preparing addressable 8 x 12 microsensor array chips was demonstrated, providing an independent and reproducible spot array.     

New synthetic routes to the kainoids: a synthesis of kainic acid and its analogues

New approaches to the synthesis of kainic acid and its analogues are presented. Two distinctly different approaches are described; the former utilised an intermolecular nitrile oxide addition to a homochiral substrate to furnish epikainate models and the second utilised amino acid chemistry to secure kainic acid.     

Changes in the characteristics of a high-filled polymeric compound in the area of its fastening to a rigid multilayer substrate

Changes in the characteristics of a high-filled polymeric compound are investigated in the area of its fastening to a rigid multilayer substrate. It is established that, in the fastening area, an increase in the amount of the rubber sol-fraction and the content of the high-filled polymeric compound plasticizer is observed, as well as an increase in the modulus of elasticity and a decrease in the strain of the HFPC.     

Needle-type concentrator and its application to the determination of pollutants

A new method for impurity concentration in gas and liquid media using a needle type microconcentrator was developed. The main advantage of this method using microconcentrators is its efficiency and simplicity. Our work was stimulated by Pawliszyn's solid-phase microconcentration method.     

有机聚合物整体柱及其在生物大分子色谱分离中的应用

整体柱是近年发展起来的一种新型的色谱分离分析技术，与传统的色谱柱相比，具有制作简单、背压低、分辨率高、柱容量大等优点。本文介绍了有机聚合物整体柱及其在生物大分子色谱分离领域的应用，分析了其存在的问题，并展望了其应用前景。     

乙炔与氢化锂单体和二聚体加成反应中的HOMO-HOMO相互作用

用前线分子轨道分析乙炔与氢化锂单体和二聚体加成反应的过渡态.结果表明,在其反应过渡态形成过程中,反应物间的HOMO-LUMO和HOMO-HOMO相互作用均起重要作用.较强烈的HOMO-HOMO相互作用与过渡态附近反应由亲电性加成向亲核性加成转变相关,该反应特性的转变又与LiH中的氢原子在反应中的电荷转移相关.     

Simple HPLC-UV determination of nucleosides and its application to the authentication of Cordyceps and its allies

A simple HPLC-UV method combined with a simple extraction procedure of nucleosides (adenosine, cordycepin, 2'-deoxyadenosine, guanosine and uridine) was developed and applied to the authentication of Cordyceps and its allies. The separation was performed on a C(18) column by isocratic elution with acetonitrile-water, and UV detection at 260 nm. The amounts of adenosine, cordycepin, 2'-deoxyadenosine, guanosine and uridine in Cordyceps were 0.28-14.15, 0.006-6.36, 0.01-0.14, 0.68-14.79 and 0.19-20.29 mg/g, respectively. Among the nucleosides studied, cordycepin was characteristically included in Cordyceps militaris (L.) Link. (CM), which is one of key Cordyceps allies, and might be a good marker for authenticating CM. The ratio of nucleosides to adenosine contents in Cordyceps seemed to be a useful marker for authentication and quality control of Cordyceps.     

On the electrostatic properties of papain in relation to its enzymatic activity

The electrostatic potentials and fields created by the thiol protease, papain, are computed on the surface envelope of the protein and in the region of its active site using a technique based on accurate multipole expansions of the electron density of appropriately chosen subunits. The effect of binding counterions to the protein is considered specifically. The possible role of the “electrostatic environment” in the functioning of the enzyme is discussed and it is found that this environment can favor the proton transfer between Cys 25 and His 159 that is thought to initiate its mode of action, but that the reasons for this appear more complicated than earlier models would suggest.     

Approaches to the solvation of the molecular probe N-methyl-6-quinolone in its excited state

The molecular probe N-methyl-6-quinolone (MQ) gives experimental access to its local chemical environment, e.g. inside a biomolecule. Using ab initio molecular dynamics (MD), it is possible to simulate the time evolution of the Stokes shift as a function of the actual atomistic coupling to the surrounding hydrogen bond network and thus obtain a comprehensive view of the local environment. In contrast to ground state ab initio MD simulations, the choice of a method for excited state MD is nontrivial. Here, we develop a simple and accurate model for the solvation dynamics of MQ in its first excited state.     

Mid-infrared second-order susceptibility of alpha-quartz and its application to visible-infrared surface sum-frequency spectroscopy

We provide the first account of the second-order susceptibility of quartz down to 10 mum (1000 cm(-1)) and show how this data may be used along with the sum-frequency response of an amorphous gold surface to elucidate the nonlinear susceptibility of any material in the mid-infrared region. Crystalline quartz is an established material for use in second-harmonic and sum-frequency generation studies of new systems, on account of its well-characterized linear and nonlinear optical properties. Previous knowledge of its nonlinear susceptibility has been limited to its transparent region, wavelengths shorter than about 3 mum. Longer wavelength chi((2)) values for quartz are particularly important for techniques such as vibrational sum-frequency spectroscopy which are expanding into the mid-IR with the increasing availability of widely tunable infrared laser sources.     

A vidicon spectrometer and its application to the analysis of gold

A TV camera which is attached to a monochromator, is used for the photoelectric measurement of optical spectra. The properties of this detection system and its application to the analysis of trace metals in high purity gold with d.c. arc excitation are described. Problems which originate from this application are discussed and results are compared with those obtained with different analytical methods.     

Overview of the synthetic routes to sildenafil and its analogues

The biological and medicinal properties of sildenafil and its analogues have prompted enormous research aimed at developing synthetic routes to these heterocycles. This review focuses on the chemical properties associated with this system.     

Overview of the synthetic routes to tadalafil and its analogues

The biological and medicinal properties of tadalafil and its analogues have prompted enormous research aimed at developing synthetic routes to these heterocyclic analogues. This review focuses on the chemical properties associated with this system.     

On the fluorescence of methyl salicylate: the significance of its IMHB

The two forms of methyl salicylate bearing an intramolecular hydrogen bond (IMHB) are responsible for the three fluorescence emissions produced by this compound on electronic excitation in inert media. Whereas the form possessing an IMHB between its hydroxyl group and ether oxygen undergoes no excited state intramolecular proton transfer (ESIPT) in its first excited electronic state, that with an IMHB involving the carbonyl oxygen exhibits ESIPT with near-unity efficiency. Whereas the former species exhibits standard photophysical behaviour, the latter species exhibits two fluorescence emissions from the same electronic excited state; a photophysical scheme is proposed, which brings together all the available photophysical evidence for methyl salicylate in inert media.     

Electrophilically catalyzed splitting of dimethoxyamine to methoxynitrenium ion and its reactions

The electrophilically catalyzed splitting of the N-0 bond of dimethoxyamine results in the formation of methoxynitrenium ion, which in reaction with nucleophiles exhibits properties of an ambidentate cation — it methylates rigid nucleophiles and epiminates olefins. The stereospecificity of the addition of the nitrenium ion to cis-2-butene indicates its existence in a singlet electronic ground state.Translated from Izvestiya Akademii Nauk SSSR, Seriya Khimicheskaya, No. 6, pp. 1384–1389, June, 1990.     

From cyclic polyenes to carbohydrates: synthesis of the hexose sugar β-allose and its 2C-branched homologue from cyclooctatetraene

In an unconventional but interesting synthetic enterprise, the commercially available hydrocarbon cyclooctatetraene (COT) has been elaborated to the rare hexose sugar (dl)-β-allose and its 2C-branched analogue. The synthetic sequence delineated here is notable for its high regio- and stereoselectivity and is flexible enough to enable access to polyoxygenated systems, hexose sugars, and their siblings from a cyclic polyene precursor.     

Facile synthesis of 9-azajulolidine and its application to post-Ullmann reactions

This Letter describes the efficient synthesis of 9-azajulolidine from readily available reagents and its utilization as an effective electron-rich ligand for post-Ullmann-type reactions, that is, for C(aryl)-heteroatom (N, O, S) bond formation reactions, with dramatically enhanced reaction rates.