Effect of 3-O-octanoyl-(+)-catechin on the responses of GABA(A) receptors and Na+/glucose cotransporters expressed in xenopus oocytes and on the oocyte membrane potential

Recently, 3-O-octanoyl-(+)-catechin (OC) was synthesized from (+)-catechin (C) by incorporation of an octanoyl chain into C in the light of (-)-epicatechin gallate (ECg) and (-)-epigallocatechin gallate (EGCg), which are the major polyphenols found in green tea and have strong physiological activities. OC was found to inhibit the response of ionotropic gamma-aminobutyric acid (GABA) receptors (GABA(A) receptors) and Na+/glucose cotransporters expressed in Xenopus oocytes in a noncompetitive manner more strongly than does C. OC also induced a nonspecific membrane current and decreased the membrane potential of the oocyte, and thus death of the oocyte occurred even at lower concentrations than that induced by C or EGCg. Although EGCg produced H2O2 in aqueous solution, OC did not. This newly synthesized catechin derivative OC possibly binds to the lipid membrane more strongly than does C, Ecg, or EGCg and as a result perturbs the membrane structure.     

PCB 118 and Aryl Hydrocarbon Receptor Immunoassays for Screening Dioxins in Retail Fish

The efficacy of a combination of two enzyme-linked immunosorbent assay (ELISA) kits was examined for screening the toxic equivalent (TEQ) concentrations of dioxins in retail fish. The coplanar PCB-EIA system, which is a competitive immunoassay specific for polychlorinated biphenyl (PCB) 118, was tested as a screening method for mono- ortho PCBs. The Ah immunoassay (Ah-I), which is an ELISA-based aryl hydrocarbon receptor binding assay, was analyzed for its screening ability for non- ortho PCBs, polychlorinated dibenzo- p-dioxins (PCDDs), and dibenzofurans (PCDFs). Dilution and recovery tests using purified fish extracts revealed no major interference of the matrix in the PCB-EIA and suggested that the matrix effect was minimized in the Ah-I. Finally, the results for the fish samples ( n = 20) showed a strong correlation between this method and high-resolution gas chromatography coupled to high-resolution mass spectrometry for the determination of the TEQ concentrations of mono- ortho PCBs ( r = 0.99) and non- ortho PCBs and PCDD/Fs ( r = 0.97). These data indicate that our method is suitable for screening retail fish to determine the TEQ concentrations of dioxins.     

旱稻水分利用率与碳同位素识别值之间的关系

通过温室盆栽试验 ,分析和探讨了三个水平的土壤水分条件对分蘖期和成熟期收获的旱稻(OryzasativaL .)生物量累积、水分利用率 (WUE)、植株不同部位的碳同位素识别值 (CID)的影响 ,并了解了它们之间的相互关系。水分条件包括 :饱和含水量 (W1 )、饱和含水量的 70 %(W2 )、饱和含水量的 4 0 %(W3)。结果表明当土壤水分条件从W1降低到W2时 ,分蘖期收获的生物量降低 4 5 %左右 ,成熟期收获的生物量降低 1 6 %～ 1 9%;而当从W1降低到W3时 ,分蘖期收获的生物量降低 73%左右 ,成熟期收获的生物量降低 5 5 %～ 5 7%。然而 ,根据地上部干重计算而来的WUE(WUE 地上部 )和根据全株干重计算而来的WUE(WUE 全株 )则随土壤含水量的降低而增加 ,其增幅在分蘖期为 0 .0 7～ 0 .2 8gkg-1 ,在成熟期为 0 .0 7～ 0 .4 5gkg-1 。植株的CID值变幅为 1 7.0～ 2 0 .6 ,但植株不同部位间差别显著 ,分蘖期收获的样品CID值从小到大的顺序为 :根 <最近完全伸展叶 <叶芽 <茎秆 ;而成熟期收获的样品CID值从小到大的顺序为 :籽粒 <根 <茎秆 <旗叶。随着土壤含水量的降低 ,植株所有部位的CID值亦显著减小。叶部的CID值与WUE 地上部(和WUE 全株 )之间呈一致的负相关关系。     

Evaluation and validation of a commercially available enzyme-linked immunosorbent assay for the neonicotinoid insecticide imidacloprid in agricultural samples

The performance of a commercially available microtiter plate ELISA kit for the determination of the neonicotinoid insecticide imidacloprid was evaluated for sensitivity, selectivity, influence of organic solvent used for extraction procedure, matrix interference originated from agricultural sample, accuracy, and method comparison with conventional HPLC analysis. The limit of detection for the kit (0.1 or 0.5 ng/mL) was determined. The working range (1-39 ng/mL) experimentally calculated on the basis of a criterion, which is determined as the range from I(20) to I(80), was comparable to that established by the manufacturer (1-50 ng/mL). The linearity of the standard curve based on the kit-assembled standard solutions agreed with the one based on the self-made standard solutions. Specificity studies indicate that the imidacloprid monoclonal antibody can readily distinguish the target compound from other structurally related neonicotinoid analogues and some metabolites, with the exception of clothianidin, the cross-reactivity of which was approximately 12%. To extract imidacloprid from an agricultural sample (apple) as simply and rapidly as possible, some extraction methods were examined. Consequently, the extraction method with hand-shaking for 5 min was the best among the examined methods. For the analysis of imidacloprid in apple samples, it was extracted directly with methanol and the extracts were diluted 10-fold (100-fold in the well) with water prior to ELISA analysis. No significant matrix interference was observed with the dilution factor. Recoveries of imidacloprid from fortified apple samples ranged from 87.7 to 112.0%. The results obtained with the ELISA kit correlated well with those by the reference method (conventional HPLC analysis) for apple samples (r > 0.998). These findings strongly indicate that the ELISA kit may be employed routinely for an on-site imidacloprid residue analysis of apple samples.     

Synthesis of haptens for development of antibodies to alkylphenols and evaluation and optimization of a selected antibody for ELISA development

The development of an enzyme-linked immunosorbent assay (ELISA) based on polyclonal antibodies for a class of endocrine disrupting compounds, 4-nonylphenol, is described. The parent molecule was derivatized at the ortho position of the free phenolic hydroxyl group to obtain the hapten, NP1, and it was conjugated with keyhole limpet hemocyanin, which was used as an immunogen. Four antisera were generated and screened against three coating antigens. The most sensitive ELISA from the screening tests (antiserum NP03As, 1/1000, and coating antigen NP1-BSA, 1 microg/mL) was further optimized and characterized. The influence of various physicochemical factors (organic solvent, pH, ion strength) was investigated. Methanol as the additive organic solvent was found to be the best organic solvent for the ELISA, with optimal sensitivity observed at a concentration of 5%. The ELISA parameters were changed at more acidic or basic pH values, whereas higher ionic strengths strongly suppressed the I(50) value and the maximum absorbance. The most sensitive ELISA for 4-nonylphenol exhibited an I(50) value of 38.6 +/- 5.5 microg/L, with a dynamic range from 12 to 350 microg/L, and the lower limit of detection was 7.7 +/- 1.3 microg/L. The optimized ELISA displayed no significant cross-reaction against the parent compounds, nonylphenol ethoxylates, degradation products, carboxylates, and bisphenol A, except in 4-octylphenol.     

Background level of indium and gallium in soil with special reference to the pollution of the soils from zinc and lead smelters

The mean and range of indium content in unpolluted soils (Humic Andosols, Eutric Fluvisols, and Mollic Gleysols) was found to be 0.037 (0.016–0.078) μg/g dry wt. In soils polluted by common heavy metals, elevated values of indium up to 1.92 μg/g dry wt were observed together with an increase of cadmium, zinc, and lead. In contrast to indium, gallium content in polluted soils was nearly the same as that in unpolluted soils throughout the profiles. The mean and range of gallium content in all unpolluted and polluted samples was 14.2 (12.8–16.3) μg/g dry wt. The indium and gallium contents in 4 Canadian reference soils were also analyzed.     

Formation of methyl mercaptan in paddy soils I

It is well known that methyl mercaptan is porduced by the microbiological decomposition of methionine1),2),3). According to Kondo ⁴⁾ and Onitake ¹⁾not only hydrogen sulfide, but also methyl mercaptan were produced from cystine by E. coli and Proteus vulgaris in the medium containing one of glucose, lactose, sucrose, glycerin or histidine. Moreover, Onitake ¹⁾ found that methyl mercaptan was produced by the action of E. coli in the medium containing hydrogen sulfide and a trace of ethyl alcohol, and that evolution of methyl mercaptan began only 5 minutes after the start of experiment in the medium containing methionine, but it began after 12hrs in the medium containing 1-cystine and glucose. According to Birkinshaw, Findlay and Webb⁵⁾ methyl mercaptan was found in the medium containing glucose, sulfate and other mineral salts, inoculated by Schizophyllum commune. In the same cultural condition as given above, methyl mercaptan, dimethyl sulfide, and dimethyl disulfide were detected by Challenger and Chartons ⁵⁾ from the data presented above, in addition to microbiological formation of methyl mercaptan from methionine, the possibility cannot be excluded of methyl mercaptan formation by microbes from cystine, sulfate or hydrogen sulfide in the medium containing one of organic compounds such as sugars, glycerin, histidine and ethyl alcohol, etc.     

Application of N2/Ar ratio method for N2 fixation studies in submerged soil

Abstract

Recently there has been developments in the measurement of N₂ fixation due mainly to the C₂H₂ reduction method (1). This method, however, has several disadvantages, especially for submerged soil, and the estimated amount of fixed N₂ on the basis of the C₂H₂ reduction activity is not very reliable. The tracer ¹⁵N₂ technique which gives a reliable estimation of the fixed N₂ is too expensive for common use. Development of an alternative method suitable for submerged soil would therefore be desirable. The present authors expected that the measurement of the ratio N₂/Ar in the soil solution might provide advantages for the estimation of the fixed N₂ in submerged soil.     

Role for piRNAs and noncoding RNA in de novo DNA methylation of the imprinted mouse Rasgrf1 locus

Genomic imprinting causes parental origin-specific monoallelic gene expression through differential DNA methylation established in the parental germ line. However, the mechanisms underlying how specific sequences are selectively methylated are not fully understood. We have found that the components of the PIWI-interacting RNA (piRNA) pathway are required for de novo methylation of the differentially methylated region (DMR) of the imprinted mouse Rasgrf1 locus, but not other paternally imprinted loci. A retrotransposon sequence within a noncoding RNA spanning the DMR was targeted by piRNAs generated from a different locus. A direct repeat in the DMR, which is required for the methylation and imprinting of Rasgrf1, served as a promoter for this RNA. We propose a model in which piRNAs and a target RNA direct the sequence-specific methylation of Rasgrf1.