Evaluation of Intestinal Absorption of Dietary Halocynthiaxanthin,a Carotenoid from the Sea Squirt Halocynthia roretzi

Halocynthiaxanthin is an acetylenic carotenoid mainly found in Halocynthia roretzi. To date, several bioactivities of halocynthiaxanthin have been reported, but its mechanism of digestion and absorption in mammals has not been studied yet. In this study, we evaluated the intestinal absorption of halocynthiaxanthin in mice. The halocynthiaxanthin-rich fraction was prepared from the tunicate Halocynthia roretzi. Mice were orally administered the fraction at a dose of 5 mg/kg body weight. The halocynthiaxanthin levels in the plasma, liver, and small intestine, were quantified using HPLC-PDA, 1, 3, 6, and 9 h after ingestion. The halocynthiaxanthin-rich fraction mainly consisted of the all-trans form and a small amount of cis forms. These three isomers were detected in the plasma of mice 3 h after ingestion. Time-course changes after the ingestion of this fraction were found, with cis isomers being more abundant than the all-trans isomer in the mouse plasma and liver. In the small intestine, however, the all-trans isomer was primarily detected. The possibility that cis isomers might be absorbed rapidly from the small intestine cannot be denied, but our results suggest that dietary all-trans-halocynthiaxanthin might be isomerized to the cis isomer after intestinal absorption.     

Effects of patch cutting on leaf nitrogen nutrition in hinoki cypress (Chamaecyparis obtusa Endlicher) at different elevations along a slope in Japan

Leaf nitrogen nutrition of hinoki cypress (Chamaecyparis obtusa Endlicher) was investigated at three positions along a slope over a period of 3 years. At each slope position, nitrogen properties were compared in patch-cut plots (0.06–0.09 ha) and uncut control plots (0.04 ha). Nitrogen cycling at the lower slope was characterized by a higher rate of soil nitrogen mineralization, and higher nitrogen concentration in fresh leaves and leaf-litter. The soil nitrogen mineralization rate and fresh-leaf nitrogen concentration in the patch-cut plots were higher than those in the control plots. However, leaf-litter nitrogen concentration did not differ between the patch-cut and control plots. The results suggest that slope position strongly affects leaf nitrogen nutrition of hinoki cypress and soil nitrogen availability. By contrast, patch cutting does not affect leaf-litter nitrogen concentration. These findings indicated that hinoki cypress would not enhance forest nitrogen cycling through changes in leaf-litter nitrogen concentration after patch cutting.     

Relation between pathogenicity and genetic variation within <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis>

The relation between diversity of pathogenicity on clubroot-resistant (CR) cultivars of Chinese cabbage (Brassica rapa subsp. pekinensis) bred in Japan and DNA polymorphisms in 17 populations of Plasmodiophora brassicae from cruciferous plants was examined by inoculation tests and random amplified polymorphic DNA (RAPD) analysis using 18 arbitrary primers. Four pathotypes (A–D) were identified after inoculation of six CR cultivars of Chinese cabbage in the 17 populations from cruciferous crops. A relatively high level of genetic diversity was also detected among these populations in the RAPD analysis. Although the four pathotypes could not be clearly differentiated using the RAPD data, most populations of three pathotypes had a consistent location on the dendrogram. All pathotype B (virulent on five cultivars except Utage 70) and D (avirulent on all cultivars) populations, which were common in incompatible interactions with cv. Utage 70, were located in a single subcluster. All five pathotype C populations (virulent only on cv. Utage 70) except for one population grouped in another single subcluster. Because four pathotype A populations (virulent on all six cultivars, races 4 and 9) fell in different subclusters, the populations may be genetically polyphyletic. Populations from cruciferous weed Cardamine flexuosa differed remarkably from those from cruciferous crops in pathogenicity on common cultivars of Chinese cabbage and turnip and C. flexuosa, but they grouped in a single cluster with all race 9 populations from crops. Race 9 populations from crops may thus be closely related to populations from the weed rather than to races 1 and 4 from crops.     

Paraticulate metals in waters of a typical irrigation river: The River Sakuragawa,Japan

The particulate metallic elements in waters of a water system of the River Sakuragawa and Lake Kasumigaura were measured by the multielement analyses using particle-induced X-ray emission. The heavy metal particulates for environmental standards were less than 0.0001 ppm for Cd, 0.00037 ppm for Cr, and 0.00078 ppm both for Hg and Pb in the river waters. The predominant metal particles in the waters were Fe, Mn, and Zn.     

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.     

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.     

Time-course comparison of pulmonary inflammation induced by intratracheal instillation of four different nickel oxide nanoparticles in male Fischer rats

Occupational exposure to nickel oxide (NiO) is an important cause of respiratory tract cancer. Toxicity is known to be associated with the dissociated component, i.e. nickel (II) ions. To address the relationship between physicochemical properties, including solubility in artificial lysosomal fluid, of NiO and time-course changes in the pulmonary response, we conducted an intratracheal instillation study in male Fischer rats using four different well-characterized NiO products, US3352 (NiO A), NovaWireNi01 (NiO B), I small particle (NiO C), and 637130 (NiO D). The NiOs were suspended in purified water and instilled once intratracheally into male F344 rats (12 weeks old) at 0 (vehicle control), 0.67, 2, and 6 mg/kg body weight. The animals were euthanized on days 3, 28, or 91 after instillation, and blood analysis, bronchoalveolar lavage fluid (BALF) testing, and histopathological examination were performed. The most soluble product, NiO B, caused the most severe systemic toxicity, leading to a high mortality rate, but the response was transient and surviving animals recovered. The second-most-soluble material, NiO D, and the third, NiO A, caused evident pulmonary inflammation, and the responses persisted for at least 91 days with collagen proliferation. In contrast, NiO C induced barely detectable inflammation in the BALF examination, and no marked changes were noted on histopathology. These results indicate that the early phase toxic potential of NiO products, but not the persistence of pulmonary inflammation, is associated with their solubility.     

Reduced glutathione decreases energy expenditure in chicks exposed to separation stress

Recently, we reported that intracerebroventricular (i.c.v.) injection of reduced glutathione (GSH) induces hypnotic and sedative effects under an acute stressful condition in chicks. However, no information is available on the effects of GSH on energy expenditure (EE) under stressful conditions. The purpose of the present study was to clarify whether i.c.v. injection of GSH affects EE of neonatal chicks, and whether EE is correlated with behavioral changes after isolation-induced stress. The EE was rapidly decreased by i.c.v. injection of GSH, but was increased 27 min after injection. This change in EE was correlated with behavioral changes in which GSH induced hypnotic and sedative effects shortly after injection, followed by a period in which activity increased. The present study demonstrates that central GSH initially causes lowered EE through hypnotic and sedative effects under an acute stressful condition in chicks.