Review of enzyme-linked immunosorbent assays (ELISAs) for analyses of neonicotinoid insecticides in agro-environments.

Immunoassay is a promising method that is suitable for rapid and simple analyses of pesticides, which are likely to persist at a trace level in agro-environments, including agricultural products, soil, and water. Particularly, enzyme-linked immunosorbent assay (ELISA) has wide application to development of analytical methods for pesticide residues because it can very sensitively and very accurately determine them in samples. This paper presents a review of the fundamental analytical performance, a device for the sample pretreatment methods before determination, and cases of applications to various samples on ELISA methods that have been developed for detection of neonicotinoid insecticides in food or environmental matrices. The reviewed ELISAs can be ranked as quantitative, rapid and simple analytical methods for single analytes. The recognition of ELISA as an analytical methodology for pesticide residues is expected to advance rapidly in the future.

Rapid and simple analysis of pesticides persisting on green pepper surfaces swabbing with solvent-moistened cotton.

A rapid and simple nondestructive extraction (NDE) method that includes wiping off of systemic neonicotinoid insecticides has been developed to streamline sample pretreatment procedures conducted before chromatographic determination. Pesticide residues were extracted from green pepper surfaces by swabbing them with absorbent cotton moistened with acetone or acetonitrile. After spraying of pesticides, the extraction rate decreased gradually, except for thiacloprid. Presumably, extraction rates depend on the physicochemical properties of pesticides, especially water solubility. It was thought that the applicability of the proposed method greatly depended on the systemic speed of each pesticide, and water solubility was placed as the index that was important to making certain. Direct analysis of some insecticides persisting on sample surfaces has been possible only by extraction before chromatographic determination. These findings indicate strongly that the proposed NDE method has collateral conditions, but it appears promising for on-site pretreatment for pesticide residue analysis.

MRT letter: In situ observation method for microstructural changes of steel during hot deformation.

We report on the result of an in situ method for observing microstructural changes during hot deformation. The observation of microstructural changes of steel at 1,473 K under tensile strain is demonstrated using the reported method. The development of deformed structures and the formation of a new grain boundary, which subsequently moved with increased strain, were clearly observed. The effectiveness of this method was confirmed by the results of several examples.

Propanil and swep inhibit 4-coumarate:CoA ligase activity in vitro.

4-Coumarate:CoA ligase (4CL, EC 6.2.1.12) in the phenylpropanoid pathway in plants has attracted interest as a novel target for developing effective plant growth inhibitors (PGIs). In a previous study in which the 4CL inhibitory activity of 28 existing herbicides was investigated using an optimized in vitro screening assay, 4CL activity was found to be strongly inhibited by propanil and swep at 100 microM. Here, further experimental evidence is provided to substantiate the previous result. Using 4-coumaric acid as substrate, tobacco 4CL activity was inhibited by propanil or swep in a concentration-dependent manner, with 50% inhibition concentrations (I(50)) of 39.6 and 6 microM respectively. These herbicides also exhibited uncompetitive inhibition towards 4-coumaric acid. Furthermore, 4CLs from several plant species were inhibited by the herbicides within a range from 1 to 50 microM. It is proposed that these herbicides have another site of action as a result of the inhibition of 4CL in the phenylpropanoid pathway, and this enzyme represents a new target site for the development of PGI.

An in vitro screening assay to discover novel inhibitors of 4-coumarate:CoA ligase.

4-Coumarate:CoA ligase (4CL, EC 6.2.1.12) exists only in plants and plays an important role in the phenylpropanoid pathway. Identification of inhibitors targeting 4CL provides a novel approach for developing effective plant growth inhibitors (PGIs). The full-length gene of tobacco 4CL (Nt4CL1) was cloned and expressed in Escherichia coli Cast & Chalm. The recombinant 4CL protein was extracted and purified by several purification steps including gel-filtration and anion-exchange chromatography. 4CL activity assay was miniaturized and optimized using a 96-well microplate and a reader. Among 28 existing herbicides, propanil and swep strongly inhibited in vitro 4CL enzyme activity, and they were selected for further studies. The process of this assay can be developed into a high-throughput screening system of PGI targeting 4CL in the phenylpropanoid pathway.