Irreversible and Reversible Deactivation of Bilirubin Oxidase by Urate

Oxygen is electroreduced to water on a carbon cathode coated with wired bilirubin oxidase in a pH 7.4 0.15 M NaCl phosphate buffer solution at 37 °C at much lesser polarization than it is on a pure platinum cathode in 0.5 M H₂SO₄. While the wired bilirubin oxidase cathode operates for over a week in the aerated or oxygenated buffer solution, it is degraded rapidly when in serum. We reported earlier that in the presence of O₂ an intermediate product of the electrooxidation of urate, which is a normal serum component, irreversibly damages the wired bilirubin oxidase and also reported that the electrocatalyst is irreversibly damaged, in the absence of urate, when it is brought, by disconnecting the electrode, to the O₂/H₂O half cell potential at pH 7.4. Here we report that a) dissolved bilirubin oxidase is irreversibly and rapidly damaged by urate in the presence of O₂; and b) that the immobilized wired bilirubin oxidase electrocatalyst is not only irreversibly deactivated by urate in the presence of O₂ in a few hours, but is initially reversibly deactivated, in 1 min or less, by the urate in the presence of O₂.     

Visual detection of cysteine and homocysteine

The determination of cysteine and homocysteine levels is of great current interest for the monitoring of desease states. A new colorimetric method for the simultaneous detection of l-cysteine and l-homocysteine has been developed. A fluorescein derivative reacts with the above amino acids, producing their respective thiazolidines resulting in color changes. Interference from other amino acids and proteins is minimal.     

PHYSIOLOGICAL RELATIONSHIPS BETWEEN PHYTOCHROME EFFECTS ON INTERNODE EXTENSION GROWTH AND DRY MATTER ACCUMULATION INLIGHT-GROWN MUSTARD

The physiological relationships between the effects of phytochrome photoequilibrium (Pfr/P) on internode extension growth and dry matter accumulation were investigated in white light (WL)-grown Sinapis alba L. seedlings. After 11 days under continuous WL, the seedlings were exposed: (a) to pulses of light providing different Pfr/P, followed by 24 h darkness (D); (b) to pulses of light providing different Pfr/P, followed by 3 h D and 24 h continuous WL; (c) to continuous WL with or without supplementary far-red light (to reduce Pfr/P); or (d) to pulses of light providing different Pfr/P followed by D, in factorial combination with either water or a saturating (0.2 M) sucrose solution applied to one of the leaves. In D (“a” and “d”) low, compared to high Pfr/P increased both internode extension growth and dry weight to the same extent. Under WL (“b” and “c”) low PfrlP promoted internode extension growth but had no proportional effects on internode dry weight. Sucrose promoted internode extension growth with a lag of at least 8 h (compared to the rapid effect of low Pfr/P) and did not reduce the effect of low Pfr/P. These results indicate that Pfr/P effects on internode extension growth are not the consequence of changes in photoassimilate translocation from the leaves. Under WL, PfdP effects on internode length occur partially at the expense of internode dry matter per unit length.     

Isolierung,Konstitution und Synthese der (R)-(−)-Eucominsäure

From the bulbs of Eucomis punctata L'Hérit. (Liliaceae) and of a hitherto undefined species of Eucomis a new optically active phenolic carboxylic acid, eucomic acid, was isolated. Structure 1 was assigned on the basis of chemical and spectral evidence. The absolute configuration of eucomic acid was determined by its correlation with piscidic acid ((2 R, 3 S)-2-(4′-hydroxybenzyl)-tartaric acid) ( 8 ). Consequently, eucomic acid is (R)-(?)-2-(4′-hydroxybenzyl)-malic acid ( 1 ). For the stereospecific synthesis, methyl cis-p-methoxybenzylidene-succinic acid ( 22 ) was transformed into the γ-lactone 24 which, by catalytic hydrogenolysis, yielded (±)-2-(4′-hydroxybenzyl)-malic acid 1-methyl ester ( 27 ). Resolution with (?)-quinine led to the enantiomeric acids 29 and 30 . The methyl ester of the levorotatory enantiomer 30 was identical with the dimethyl ester 3 of 4′-O-methyl-eucomic acid.     

Assignment of the relative configuration of spiro[4.5]decanes by 13C, 15N and 17O NMR

The relative configuration of 11 1,4-diazaspiro[4.5]decanes (1a-1j and 1m), 15 1,4-oxazaspiro[4.5]decanes (2a-2o) and 10 1,4-dioxaspiro[4.5]decanes (3a-3n) substituted at the 2-, 6-, 7- or 8-position by a methyl group or using the tert-butyl group as a model for the ananchomeric structure is reported. The relative stereochemistry was analyzed by 1H, 13C, 15N and 17O NMR and all isomers present were characterized spectroscopically. Compounds with a methyl group in the six-membered ring show a chair conformation preference with the methyl group in the equatorial position. Compounds with one or two nitrogens exhibit a tautomeric equilibrium between the imine-diazolidine forms, as demonstrated by IR and 13C NMR.     

Stille-type cross-coupling-An efficient way to various symmetrically and unsymmetrically substituted methyl-bipyridines: toward new ATRP catalysts

Various mono- and disubstituted 2,2'-bipyridines were synthesized in high yields and multigram scales utilizing Stille-type coupling procedures. The corresponding bromo-picoline and tributyltin-picoline building blocks were prepared from commercially available amino-picoline compounds. As first examples of metal complexes, 4,5'-dimethyl-2,2'-bipyridine was reacted with copper(II) and iron(II) ions and investigated as catalyst in ATRP.     

Determination of oxidized and reduced glutathione, by capillary zone electrophoresis, in Brassica juncea plants treated with copper and cadmium

A rapid method of capillary zone electrophoresis is described to determine the oxidized (GSSG) and reduced (GSH) form of glutathione in plant tissue. In order to separate both analytes in a fused-silica capillary, the pH and composition of the electrolyte solution were optimized. The electrolyte composition was 100 mmol/L, borate 25 mmol/L Tris, and 0.2% w/v metaphosphoric acid (MPA), pH 8.2. Some instrumental conditions used to run the samples were hydrostatic injection for 30 s, 30 kV applied voltage, and UV detection (185 nm) at 25 degrees C. Linearity and useful range obtained for the calibration curves were optimum, with correlation coefficients about 0.999 in the 0-120 micromol/L range. The migration time was highly reproducible, less than 5 min being afforded to run a sample. Electrolyte buffer and samples required a careful pH control for optimal separation of both analytes. This aspect constitutes a critical analytical step when acids are used in the procedure for sample preparation. Simultaneous analysis of GSH and GSSG may provide a useful tool for comparative studies of plants in order to select those species with a potential capacity for detoxification from toxic elements or those appearing promising from phytoremediation for these elements.     

Transport in ceria electrolytes modified with sintering aids: effects on oxygen reduction kinetics

Small (2 mol%) cobalt oxide additions to ceria-gadolinia (CGO) materials considerably improve sinterability, making it possible to obtain ceramics with 95–99% density and sub-micrometre grain sizes at 1,170–1,370 K. The addition of Co causes a significant shift of the electrolytic domain to lower pO₂. This modification to the minor electronic conductivity of the electrolyte material has influence on the cathodic oxygen reduction reaction. The impedance technique is shown to provide information not only about polarisation resistance, but also about the active electrode area from analysis of the current constriction resistance. It is demonstrated that this current constriction resistance can be related to the minor electronic contributions to total conductivity in these materials. A simple imbedded grid approach gives control of the contact area allowing the properties of the electrolyte materials to be studied. A much lower polarisation resistance for the Co-containing CGO electrolyte is observed, which can be clearly attributed to an increased three-phase reaction area in the Co-containing material, as a consequence of elevated p-type conductivity.     

Annulative π-extension of BODIPYs made easy via gold(i)-catalyzed cycloisomerization

Here we report gold(i)-catalyzed cycloisomerization as a new powerful synthetic tool for the preparation of π-extended BODIPY derivatives. The catalytic system PPh^F₃AuCl/AgSbF₆ enables the synthesis of [b]-[2,1]naphtho-fused-BODIPYs (2a–2c) under mild conditions, in excellent yields and short reaction times. The reaction is totally regioselective to the 6-endo-dig product and for the α-position of the BODIPY, which is both the kinetically and thermodynamically favored pathway, as supported by the free energy profile calculated by means of Density Functional Theory (DFT). Moreover, this methodology also allows the synthesis of two new families of [b]-aryl-fused-BODIPYs, namely, [3,4]phenanthro- (2e and 2f) and [1,2]naphtho-fused (2g) BODIPYs. Their molecular and electronic structures were established by NMR and UV-vis spectroscopies as well as single-crystal X-ray diffraction analysis. As can be noted from the X-ray structures, 2a, 2e and 2g present interesting structural differences at both the molecular and packing level. Interestingly, despite being isomers, the UV/vis spectra of 2a and 2g revealed significant differences in their electronic structures. The origin of this finding was studied by Time-Dependent DFT calculations. Calculated DFT Nuclear Independent Chemical Shift (NICS(0)) values also supported the different electronic structures of 2a and 2g.

Here we report gold(i)-catalyzed cycloisomerization as a new powerful synthetic tool for the preparation of π-extended BODIPY derivatives.     

NADH Electrooxidation Using Bis(1,10-phenanthroline-5,6-dione) (2,2'-bipyridine)ruthenium(II)-Exchanged Zirconium Phosphate Modified Carbon Paste Electrodes

We present a carbon paste electrode (CPE) modified using the electron mediator bis(1,10‐phenanthroline‐5,6‐dione)(2,2′‐bipyridine)ruthenium(II) ([Ru(phend)₂bpy]²⁺) exchanged into the inorganic layered material zirconium phosphate (ZrP). X‐Ray powder diffraction showed that the interlayer distance of ZrP increases upon [Ru(phend)₂bpy]²⁺ intercalation from 10.3 Å to 14.2 Å. The UV‐vis and IR spectroscopies results showed the characteristic peaks expected for [Ru(phend)₂bpy]²⁺. The UV‐vis spectrophotometric results indicate that the [Ru(phend)₂bpy]²⁺ concentration inside the ZrP layers increased as a function of the loading level. The exchanged [Ru(phend)₂bpy]²⁺ exhibited luminescence even at low concentration. Modified CPEs were constructed and analyzed using cyclic voltammetry. The intercalated mediator remained electroactive within the layers (E°′=–38.5 mV vs. Ag/AgCl, 3.5 M NaCl) and electrocatalysis of NADH oxidation was observed. The kinetics of the modified CPE shows a Michaelis–Menten behavior. This CPE was used for the oxidation of NADH in the presence of Bakers' yeast alcohol dehydrogenase. A calibration plot for ethanol is presented.