Decontamination effect of electrolysed NaCl solutions on carp

AIMS: To evaluate the efficacy of electrolysed NaCl solutions (EW) for disinfecting bacterial isolates from carp, and the potential application of EW to reducing the bacterial load in whole carp and carp fillets. METHODS AND RESULTS: EW was produced by using a two-compartment batch-type electrolysed apparatus. Pure cultures (in vitro), whole carp (skin surface) and carp fillets were treated with EW to detect its antimicrobial effects. The anodic solution [EW (+)] completely inhibited growth of the isolates. Furthermore, dipping the fish samples in EW (+) reduced the mean total count of aerobic bacteria on the skin of whole carp and in fillets by 2.8 and 2.0 log(10), respectively. The cathodic solution [EW (-)] also reduced growth of the isolates from carp by ca 1.0 log(10). Moreover, the total counts of aerobic bacteria in whole carp (on the skin) and fillets were reduced by 1.28 and 0.82 log(10), respectively. CONCLUSIONS: EW (+) has a strong bactericidal effect on bacteria isolated from carp. SIGNIFICANCE AND IMPACT OF THE STUDY: Treatment with EW (+) could extend the shelf life of these fish.     

γ‐Aminobutyric acid quantification in small volume biological samples through enzymatically induced electrochemiluminescence

γ‐Aminobutyric acid (GABA) is a well‐known neurotransmitter that regulates inhibitory neurotransmission in the mammalian central nervous system and participates in several processes outside the brain. A reliable quantification method is needed to determine its role in different physiological and pathological conditions. However, GABA measurements have several challenges because GABA is neither fluorescent nor electroactive, and it is difficult to detect using enzymatic reactions because no oxidases or dehydrogenases have been identified. Several methods have been developed to quantify GABA concentrations based on the instrumentation available, the sensitivity required, and the volume of samples analyzed. Most of these methods use high‐performance liquid chromatography (HPLC). Here, we describe a method for quantifying GABA concentrations in small volume samples using enzymatically‐induced electrochemiluminescence with the well‐known GABAse complex, which produces glutamate for use in a luminescent reaction with glutamate oxidase and luminol in an electrochemiluminescence cell. The luminescence obtained was proportional to the GABA concentrations in the micromolar range (1–1000), with linear r² values > 0.95. GABA standards were treated with the enzymatic reactors to generate glutamate (Glu), which was measured simultaneously with an HPLC technique, to validate this new procedure. The assay was further used to determine GABA concentrations in hippocampal extracts. This alternative may be used to quantify GABA levels in fluid samples, such as microdialysates, other perfusates and tissue extracts. Thus, the method presented here is a good alternative for monitoring GABA levels with good sensitivity compared with the traditional methods that are still in use.     

Schistosoma mansoni:The Circulating Cathodic Antigen Forms an Abundant Product of 41/42 kDa in the Urine of Infected Patients

Abdeen, H. H., Attallah, A.-F. M., El-Mohamady, H. I., Harrison, R. A., and Mansour, M. M. 1998.Schistosoma mansoni: The circulating cathodic antigen forms an abundant product of 41/42 kDa in the urine of infected patients.Experimental Parasitology90, 286–289.     

New polyacrylamide matrices for drift-free isoelectric focusing

The major cause for pH gradient decay and cathodic drift during isoelectric focusing in polyacrylamide gels has been found to be electroendo-osmotic flow generated by fixed charges in the gel matrix. These charges have the following causes: (a) trace impurities of acrylic acid in the co-monomers; (b) covalent incorporation of catalysts (persulphate and riboflavin 5'-phosphate) as terminal groups in polyacrylamide chains; (c) hydrolysis of amide groups to acrylic acid in the gel layer underneath the cathodic filter paper strip. The result of these fixed negative charges in the matrix is a movement of counter-ions with hydration water towards the cathode (i.e. electroendo-osmosis) with concomitant drift of pH gradient and focused protein zones in the same direction. It is impossible to cure the cathodic drift by increasing the pH of the anolyte, or decreasing the pH of the catholyte, or both, as previously suggested. One way to reduce the cathodic drift efficiently is to incorporate covalently in the matrix tertiary or quaternary groups (3-dimethylaminopropylmethacrylamide) in stoichiometric amounts as compared with the negative charges.This ‘balanced’ polyacrylamide displays zero drift for at least 5000V·h, which is considered to be an ample time for equilibrium separation of protein species in isoelectric focusing.     

Multi-channel electrochemiluminescence of luminol at a copper electrode.

Multi-channel electrochemiluminescence (ECL) of luminol at a copper electrode has been studied under conventional cyclic voltammetric (CV) conditions. Compared with the ECL of luminol at other electrodes, three ECL peaks were observed at 0.30, -0.24 and -0.65 V (vs. SCE), respectively, which was also imaged by a CCD camera. The effects of potential scan direction, anodic reverse potential, the presence of N2 and O2 of the solution, the pH of the solution, the NaNO3 concentration and the potential scan rate were examined. The effect of n-alkanethiol self-assembled monolayers on copper electrodes and 20 L-amino acids, dopamine, adrenaline and noradrenaline on the ECL of luminol were also investigated. The emission spectra of various ECL peaks at different potentials demonstrated that all ECL peaks were related to the luminol reaction. The results show that the oxygen dissolved in solution and copper oxide covered on the surface of the electrode play an important role in the luminol ECL process at a copper electrode. It has been proposed that three ECL channels of luminol at a copper electrode resulted from the reactions of luminol or luminol radical electrooxidized by luminol with various electrogenerated oxygen-containing species, such as O2, OOH- and copper oxides at different potentials.     

Electrochemiluminescence determination of codeine or morphine with an organically modified silicate film immobilizing Ru(bpy)3(2+).

An ECL approach was developed for the determination of codeine or morphine based on tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)(3)(2+)) immobilized in organically modified silicates (ORMOSILs). Tetramethoxysilane (TMOS) and dimethyldimethoxysilane (DiMe-DiMOS) were selected as co-precursors for ORMOSILs, which were then immobilized on a surface of glassy carbon electrode (GCE) by a dip-coating process. Ru(bpy)(3)(2+) was immobilized in the ORMOSIL film via ion-association with poly(p-styrenesulphonate). The ORMOSIL-modified GCE presented good electrochemical and photochemical activities. In a flow system, the eluted codeine or morphine was oxidized on the modified GCE and reacted with immobilized Ru(bpy)(3)(2+) at a potential of +1.20 V (vs. Ag/AgCl). The modified electrode was used for the ECL determination of codeine or morphine and showed high sensitivity. The calibration curves were linear in the range 2 x 10(-8)-5 x 10(-5) mol/L for codeine and 1 x 10(-7)-3 x 10(-4) mol/L for morphine. The detection limit was 5 x 10(-9) mol/L for codeine and 3 x 10(-8) mol/L for morphine, at signal:noise ratio (S:N)=3. Both codeine and morphine showed reproducibility with RSD values <2.5% at 1.0 x 10(-6) mol/L. Furthermore, the modified electrode immobilized Ru(bpy)(3)(2+) was applied to the ECL determination of codeine or morphine in incitant samples.     

Studies on lysozyme modifications induced by substituted p-benzoquinones

Protein misfolding can facilitate a protein damaging process and makes it susceptible to a series of events such as unfolding, adduct formation, oligomerization, or aggregation. Loss of a protein’s native structure may result in its biological malfunction and/or cellular toxicity that could cause associated diseases. Several factors were identified for causing structural changes of a protein, however quinone-induced protein modifications received very little attention whether for amyloidal or non-amyloidal proteins. In this paper, we report our investigation on lysozyme modifications upon treatment with selected benzoquinones (BQs), utilizing fluorescence spectroscopy including anisotropy determination, UV–Vis spectroscopy, and SDS-PAGE. Lysozyme was reacted with substituted BQs in order to examine substituent effects on protein modifications. In addition, we evaluated lysozyme modifications induced by 1,4-benzoquinone in concentration-, pH-, temperature-, and time-dependent studies. Our study shows that all BQs can readily modify lysozyme in a complex manner through adduct formation, oligomerization, polymeric aggregation, and/or fibrilization. Electrochemical properties of selected BQs were monitored using cyclic voltammetry in phosphate buffered aqueous solution, and it was found that quinone reduction potentials correlate well with their reactivity trend toward lysozyme.     

Exploring the electrochemiluminescent behavior of procaterol hydrochloride in the presence of Ru(bpy)32+ and its analytical application in pharmaceutical preparation

Based on the strong enhancement effect of procaterol hydrochloride on the electrochemiluminescence (ECL) of Ru(bpy)₃²⁺ (bpy = 2,2′‐bipyridine) in an alkaline H₃PO₄–NaOH buffer solution on a bare Pt electrode, a simple, rapid and sensitive method was developed for the determination of procaterol hydrochloride. The optimum conditions for the enhanced ECL have been developed in detail in this work. Under optimum conditions, the logarithmic ECL enhancement vs. the logarithmic concentration of procaterol hydrochloride is linear over a wide concentration range of 2.0 × 10^?7 to 2.0 × 10^?4 M (r =  0.9976), with a limit of detection of 1.1 × 10^?8 M (S/N =  3), and a relative standard deviation of 2.1% (n =  7, c =  5.0 × 10^?6 M). The proposed method was applied to the determination of this drug in tablets with recoveries of 89.7%–98.5%. In addition, a possible mechanism for the enhanced ECL of Ru(bpy)₃²⁺, which is caused by ProH, has also been proposed.     

Self‐quenching in the electrochemiluminescence of Ru(bpy)32+ using ascorbic acid as co‐reactant

In this study, electrochemiluminescence (ECL) of Ru(bpy)₃²⁺ (bpy = 2,2′‐bipyridyl) using ascorbic acid (H₂A) as co‐reactant was investigated in an aqueous solution. When H₂A was co‐existent in a Ru(bpy)₃²⁺‐containing buffer solution, ECL peaks were observed at a potential corresponding to the oxidation of Ru(bpy)₃²⁺, and the intensity was proportional to H₂A concentration at lower concentration levels. The formation of the excited state *Ru(bpy)₃²⁺ was confirmed to result from the co‐reaction between Ru(bpy)₃³⁺and the intermediate of ascorbate anion radical (A⁻•), which showed the maximum ECL at pH = 8.8. It is our first finding that the ECL intensity would be quenched significantly when the concentration of H₂A was relatively higher, or upon ultrasonic irradiation. In most instances, quenching is observed with four‐fold excess of H₂A over Ru(bpy)₃²⁺. The diffusional self‐quenching scheme as well as the possible reaction pathways involved in the Ru(bpy)₃²⁺–H₂A ECL system are discussed in this study. Copyright © 2008 John Wiley & Sons, Ltd.