High-affinity monoclonal antibodies for detection of the microbial metabolite, 2-methylisoborneol

The production of 2-methylisoborneol (MIB) by certain fungi and algae can contribute musty off-flavors to foods and water supplies if uncontrolled. The goal of this research was to develop a nonsensory simple method for the detection of MIB. Anti-MIB monoclonal antibodies were produced by immunizing mice with borneol-conjugated protein and selecting positive clones with an MIB-protein conjugate. An indirect competitive immunoassay developed using this antibody had a detection limit of 0.6 microg L(-)(1) and an I(50) value of 5 microg L(-)(1). Detection was relatively specific for MIB and showed 20% cross-reactivity with borneol or isoborneol and 4-5% cross-reactivity with camphor. No cross-reactivity to geosmin was observed.     

Measurement of Atmospheric Formaldehyde and Monoaromatic Hydrocarbons using Differential Optical Absorption Spectroscopy during Winter and Summer Intensive Periods in Seoul, Korea

Ozone effects on Mediterranean forests were studied for 10 years in southern France. The aim of these studies was to understand the impact of the pollutant on foliar damage among Aleppo pine (Pinus halepensis Mill) and Arolla pine (Pinus cembra L) populations. During the summers of 2001 and 2002, ozone concentrations were measured using UV absorption analysers (4) and passive samplers (29) at two forest monitoring sites: Site 1 in the Nice coastal area for Aleppo pine and Site 2 in the Mercantour mountains for Arolla pine, in the vicinity of tree observation sites. AOT 40 values were quite high (25–33 ppm.h), especially in the Mercantour mountains, for the 5 months of measurement. Foliar damage (defoliation and discoloration) and specific symptoms (chlorotic mottle) were observed on 8 Aleppo pine and 13 Arolla pine plots. For Arolla pines, there was a significant correlation between defoliation and chlorotic mottles. There was also a correlation between mean ozone concentrations measured by passive samplers and chlorotic mottles. These correlations were less significant for Aleppo pine.     

Multi-Step Competitive Sorption and Desorption of Chlorophenols in Surfactant Modified Montmorillonite

Single- and bi-solute sorption and desorption of 2,4-dichlorophenol (2,4-DCP) and 2,4,5-trichlorophenol (2,4,5-TCP) in montmorillonite modified with hexadecyltrimethylammonium (HDTMA) were investigated using multi-step sorption and desorption procedure. Effect of pH on the multi-step sorption and desorption was investigated. As expected by the magnitude of octanol-water partition coefficient, K _ow , both sorption and desorption affinity of 2,4,5-TCP was higher than that of 2,4-DCP at pH 4.85 and 9.15. For both chlorophenols, the protonated speciation (at pH 4.85) exhibited a higher affinity in both sorption and desorption than the predominant deprotonated speciation (about 95% and 99% of 2,4-dichlorophenolate and 2,4,5-trichlophenolate anions at pH 9.15, respectively). Desorption of chlorinated phenols was strongly dependent on the current pH regardless of their speciation in the previous sorption stage. Freundlich model was used to analyze the single-solute sorption and desorption data. No appreciable desorption-resistant (or non-desorbing) fraction was observed in organoclays after several multi-step desorptions. This indicates that sorption of phenols in organoclay mainly occurs via partitioning into the core of the pseudo-organic medium, thereby causing desorption nearly reversible. In bisolute competitive systems, sorption (or desorption) affinity of both chlorophenols was reduced compared to that in its single-solute system due to the competition between the solutes. The ideal adsorbed solution theory (IAST) coupled to the single-solute Freundlich model successfully predicted bisolute multi-step competitive sorption and desorption equilibria.     

Lipoxygenase inhibitory activity of anacardic acids

6[8'(Z)-pentadecenyl]salicylic acid, otherwise known as anacardic acid (C15:1), inhibited the linoleic acid peroxidation catalyzed by soybean lipoxygenase-1 (EC 1.13.11.12, type 1) with an IC50 of 6.8 microM. The inhibition of the enzyme by anacardic acid (C15:1) is a slow and reversible reaction without residual activity. The inhibition kinetics analyzed by Dixon plots indicates that anacardic acid (C15:1) is a competitive inhibitor and the inhibition constant, KI, was obtained as 2.8 microM. Although anacardic acid (C15:1) inhibited the linoleic acid peroxidation without being oxidized, 6[8'(Z),11'(Z)-pentadecadienyl]salicylic acid, otherwise known as anacardic acid (C15:2), was dioxygenated at low concentrations as a substrate. In addition, anacardic acid (C15:2) was also found to exhibit time-dependent inhibition of lipoxygenase-1. The alk(en)yl side chain of anacardic acids is essential to elicit the inhibitory activity. However, the hydrophobic interaction alone is not enough because cardanol (C15:1), which possesses the same side chain as anacardic acid (C15:1), acted neither as a substrate nor as an inhibitor.     

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.     

Correlation between Environmental Conditions and the Distribution of Mosses Exposed to Urban Air Pollutants

Moss surveys have several advantages over conventional depositionmonitoring based on precipitation, and allow the study of regional differences and temporal trends of airborne pollution. The purposes of this study were to: (i) describe the moss species growing in one of the largest industrial cities in Asia, (ii) document edaphic and atmospheric conditions at each site, (iii) collect materials and data for a program of regular moss monitoring, previously lacking in the Far East, (iv) uncover correlations among atmospheric and edaphic factors expected to influence moss growth. As a result, a total of fifteen moss species were recorded in twenty-five sites in Seoul, Korea, and most sites had four or five species. Each species' density was affected by soil parameters, but density of Pogonatum nessii and Taxiphyllum taxirameum was also affected by CO. In contrast, SO₂ was the only environmental variablethat significantly affected the composition of moss communities. On the whole, species diversity rather than abundance of specific moss species might be a good indicator of atmospheric contamination in Seoul. This investigation will help fill the gap in knowledge of the interactions between environmental pollution and moss vegetation under field conditions.     

Influence of green tea polyphenol in rats with arginine-induced renal failure

To determine whether green tea polyphenol ameliorates the pathological conditions induced by excessive dietary arginine, green tea polyphenol was administered to rats at a daily dose of 50 or 100 mg/kg body weight for 30 days with a 2% w/w arginine diet. In arginine-fed control rats, urinary and/or serum levels of guanidino compounds, nitric oxide (NO), urea, protein, and glucose increased significantly, while the renal activities of the oxygen species-scavenging enzymes superoxide dismutase (SOD) and catalase decreased, compared with casein-fed rats. However, rats given green tea polyphenol showed significant and dose-dependent decreases in serum levels of creatinine (Cr) and urea nitrogen and urinary excretion of Cr, and they exerted a slight reduction of nitrite plus nitrate, indicating that green tea polyphenol reduced the production of uremic toxins and NO. In addition, in arginine-fed rats the urinary urea, protein, and glucose level increases were reversed by the administration of green tea polyphenol. Moreover, in rats given green tea polyphenol the SOD and catalase activities suppressed by excessive arginine administration increased dose-dependently, implying the biological defense system was augmented as a result of free radical scavenging. These results suggest that green tea polyphenol would ameliorate renal failure induced by excessive dietary arginine by decreasing uremic toxin, and NO production and increasing radical-scavenging enzyme activity.     

Peroxynitrite scavenging and cytoprotective activity of 2,3,6-tribromo-4,5-dihydroxybenzyl methyl ether from the marine alga Symphyocladia latiuscula

Peroxynitrite (ONOO(-)), formed from the reaction of superoxide (O(2)*(-)) and nitric oxide (*NO), is a cytotoxic species that can oxidize several cellular components such as proteins, lipids, and DNA. It has been implicated in diseases such as Alzheimer's disease, rheumatoid arthritis, cancer, and atherosclerosis. Due to the lack of endogenous enzymes responsible for ONOO(-) inactivation, developing a specific ONOO(-) scavenger is of considerable importance. The aim of this study was to evaluate the ability of marine natural products to scavenge ONOO(-) and to protect cells against ONOO(-). Methanolic extracts of 17 marine alga were tested for their ONOO(-) scavenging activity. Among them, Symphyocladia latiuscula showed the potent scavenging activity. CH(2)CH(2) fraction was partitioned with CH(2)CH(2) following n-hexanal extraction from the methanol extract of S. latiuscula. It was highly effective for ONOO(-) scavenging activity. Further analysis of the active fractionated extract identified 2,3,6-tribromo-4,5-dihydroxybenzyl methyl ether (TDB) as a potent ONOO(-) scavenger. The data demonstrated that TDB led to decreased ONOO(-)-mediated nitration of tyrosine through electron donation. TDB showed significant inhibition on nitration of bovine serum albumin and low-density lipoprotein by ONOO(-) in a dose-dependent manner. It also provided cytoprotection from cell damage induced by ONOO(-). TDB can be developed as an effective peroxynitrite scavenger for the prevention of the involved diseases.     

Biochemically marked lymphocytoid lines: establishment of Lesch-Nyhan cells

Two lymphocytoid cell lines have been established from a patient with the Lesch-Nyhan syndrome. The cells are deficient in hypoxanthine-guanine phosphoribosyltransferase, as demonstrated by their failure to incorporate [H(3)]hypoxanthine and by their inability to grow in medium in which they were nutritionally dependent upon exogenous hypoxanthine. This represents the first establishment of presumptively permanent human lymphocytoid cell lines that are deficient in a specific enzyme.