Application of ELISA-based kit for detecting AOZ and determining its clearance in eel tissues

Furazolidone, an antibacterial drug that was once widely used in the livestock industry and aquaculture, is now prohibited in numerous countries. It is difficult to detect residual furazolidone because it is readily metabolized in animal tissues but, by using and liquid chromatography coupled with tandem mass spectrometry, its metabolite, 3-amino-2-oxazolidinone (AOZ) can be detected. Here we describe the validity of an enzyme-linked immunosorbent assay (ELISA) kit to detect AOZ in Japanese eel Anguilla japonica tissue. ELISA is capable of detecting AOZ at 1.0 μg/kg in an eel sample with excellent accuracy and precision. Our results show that ELISA is suitable for regulatory purposes and for studying the fate of AOZ residues in eel treated with furazolidone. To measure the persistence of AOZ in eel tissues, eels (1.4–6.5g) were immersed in tanks containing 2 and 10 mg furazolidone/L for 3 h, and then maintained in a tank supplying well water for the next 160 days. The half-lives of AOZ, calculated from the linear terminal part of the excretion curve, were 25.0 days in muscle and 21.6 days in liver from fish exposed to 2 mg/L furazolidone. In the eels treated with 10 mg/L furazolidone, by contrast, high levels of AOZ were detected in liver and muscle, but the half-lives of AOZ were similar to those in fish treated with 2 mg/L furazolidone. The half-lives of AOZ in eel tissues were prolonged by the condition of low water temperature.     

Comparisons of energy balance and evapotranspiration between flooded and aerobic rice fields in the Philippines

The seasonal and annual variability of sensible heat flux (H), latent heat flux (LE), evapotranspiration (ET), crop coefficient (K_c) and crop water productivity (WP_ET) were investigated under two different rice environments, flooded and aerobic soil conditions, using the eddy covariance (EC) technique during 2008-2009 cropping periods. Since we had only one EC system for monitoring two rice environments, we had to move the system from one location to the other every week. In total, we had to gap-fill an average of 50-60% of the missing weekly data as well as those values rejected by the quality control tests in each rice field in all four cropping seasons. Although the EC method provides a direct measurement of LE, which is the energy used for ET, we needed to correct the values of H and LE to close the energy balance using the Bowen ratio closure method before we used LE to estimate ET. On average, the energy balance closure before correction was 0.72 ± 0.06 and it increased to 0.99 ± 0.01 after correction. The G in both flooded and aerobic fields was very low. Likewise, the energy involved in miscellaneous processes such as photosynthesis, respiration and heat storage in the rice canopy was not taken into consideration.Average for four cropping seasons, flooded rice fields had 19% more LE than aerobic fields whereas aerobic rice fields had 45% more H than flooded fields. This resulted in a lower Bowen ratio in flooded fields (0.14 ± 0.03) than in aerobic fields (0.24 ± 0.01). For our study sites, evapotranspiration was primarily controlled by net radiation. The aerobic rice fields had lower growing season ET rates (3.81 ± 0.21 mm d⁻¹) than the flooded rice fields (4.29 ± 0.23 mm d⁻¹), most probably due to the absence of ponded water and lower leaf area index of aerobic rice. Likewise, the crop coefficient, K_c, of aerobic rice was significantly lower than that of flooded rice. For aerobic rice, K_c values were 0.95 ± 0.01 for the vegetative stage, 1.00 ± 0.01 for the reproductive stage, 0.97 ± 0.04 for the ripening stage and 0.88 ± 0.03 for the fallow period, whereas, for flooded rice, K_c values were 1.04 ± 0.04 for the vegetative stage, 1.11 ± 0.05 for the reproductive stage, 1.04 ± 0.05 for the ripening stage and 0.93 ± 0.06 for the fallow period. The average annual ET was 1301 mm for aerobic rice and 1440 mm for flooded rice. This corresponds to about 11% lower total evapotranspiration in aerobic fields than in flooded fields. However, the crop water productivity (WP_ET) of aerobic rice (0.42 ± 0.03 g grain kg⁻¹ water) was significantly lower than that of flooded rice (1.26 ± 0.26 g grain kg⁻¹ water) because the grain yields of aerobic rice were very low since they were subjected to water stress.The results of this investigation showed significant differences in energy balance and evapotranspiration between flooded and aerobic rice ecosystems. Aerobic rice is one of the promising water-saving technologies being developed to lower the water requirements of the rice crop to address the issues of water scarcity. This information should be taken into consideration in evaluating alternative water-saving technologies for environmentally sustainable rice production systems.     

Factors affecting the magnitude of post-glacial hillslope incision in Japanese mountains

In Japanese mountains, increased post-glacial rainfall triggered gullying in bedrock and thousands of shallow failures in regolith, resulting in widespread hillslope incision. To identify factors influencing the areal extent of the post-glacial incision, fifteen rugged river basins in central and northeast Japan were surveyed. Topographical and geological properties of hillslopes were measured on geomorphological maps, digital elevation data, and geological maps. Analyses of ca. 3,000 0.5 × 0.5 km morphometric samples have shown that the magnitude of regolith failure depends mainly on slope inclination and geology, and subordinately on local storm intensity. The magnitude of bedrock gullying also depends on these three factors, but with greater dominance of storm intensity. These observations reflect the differential stability of hillslopes with varying inclination, erodibility of bedrock, and sensitivity of regolith and bedrock to change in storm intensity. The three factors also exert control on alluvial-fan development in the lower reaches, because they affect the amount of hillslope sediment supply and in turn the type of fluvial processes.     

Identification of reaction products of acylated anthocyanins from red radish with peroxyl radicals

Red radish anthocyanin extract, which consists of 12 known acylated anthocyanins, was reacted with 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) to generate peroxyl radicals under acidic pH conditions at 37 degrees C. The reaction products were isolated using preparative HPLC, and their chemical structures were determined to be p-hydroxybenzoic acid (1), 6-O-(E)-p-coumaroyl-2-O-beta-d- glucopyranosyl-alpha-d-glucopyranoside (3), p-coumaric acid (4), 6-O-(E)-feruloyl-2-O-beta-d-glucopyranosyl-alpha-d-glucopyranoside (5), and ferulic acid (6). Some products were not identified. HPLC analyses of the mixture of acylated pelargonidin isolated from red radish and AAPH revealed that the acylated pelargonidins possess the radical scavenging ability on some common sites even if the characteristics of the intramolecular acyl units are different. Degradation rates of acylated pelargonidins and the formation rates of the resulting reaction products were found to be quite different.     

Oxidative degradation of bisphenol a by crude enzyme prepared from potato

When crude enzymes prepared from some vegetables and fruits were incubated with bisphenol A (2,2-bis(4-hydroxyphenyl)propane, BPA) at 37 degrees C, BPA was oxidized by crude enzymes from potato, eggplant, and lettuce. The crude enzyme prepared from potato (Solanum tuberosum) had the strongest oxidative activity for BPA. Its optimal temperature and pH were 40-45 degrees C and 8.0, respectively. More than 95% of BPA was oxidized after the incubation with potato enzyme for 60 min. BPA gave two oxidation products besides insoluble compounds during the oxidation by potato enzyme. The oxidation products were identified to be 4[1-(4-hydroxyphenyl)-1-methyl-ethyl]-benzene-1,2-diol and 4[1-(4-hydroxyphenyl)-1-methyl-ethyl]-benzene-1,3-diol. Enzymatically oxidized BPA lost the estrogen-like activity to enhance the growth of human breast cancer (MCF-7) cells.     

The influence of barley malt protein modification on beer foam stability and their relationship to the barley dimeric alpha-amylase inhibitor-I (BDAI-I) as a possible foam-promoting protein

The foam stability of beer is one of the important key factors in evaluating the quality of beer. The purpose of this study was to investigate the relationship between the level of malt modification (degradation of protein, starch, and so on) and the beer foam stability. This was achieved by examining foam-promoting proteins using two-dimensional gel electrophoresis (2DE). We found that the foam stability of beer samples brewed from the barley malts of cultivars B and C decreased as the level of malt modification increased; however, the foam stability of cultivar A did not change. To identify the property providing the increased foam stability of cultivar A, we analyzed beer proteins using 2DE. We analyzed three fractions that could contain beer foam-promoting proteins, namely, beer whole proteins, salt-precipitated proteins, and the proteins concentrated from beer foam. As a result, we found that in cultivar A, some protein spots did not change in any of these three protein fractions even when the level of malt modification increased, although the corresponding protein spots in cultivars B and C decreased. We analyzed these protein spots by peptide mass finger printing using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. As a result, all of these spots were identified as barley dimeric alpha-amylase inhibitor-I (BDAI-I). These results suggest that BDAI-I is an important contributor to beer foam stability.     

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.     

Dual effects of reversed winter–spring temperatures on year‐to‐year variation in the recruitment of chub mackerel (Scomber japonicus)

To clarify the effects of temperature on the recruitment of chub mackerel (Scomber japonicus) in the North Pacific, we investigated the influence of winter surface temperature (WST) on spawners at the time of maturity around the spawning grounds and the influence of ambient spring temperature on larvae using estimated temperature (ET) obtained from particle tracking experiments. We found a significant positive correlation between ET approximately 10 days following hatching and the recruitment per spawning stock biomass (RPS) after 2000. The closer (more meandering) the Kuroshio Current (KC) axis was in relation to the spawning ground, the higher (lower) the spring surface temperature and the higher (lower) RPS was in the spawning ground. In contrast, WST inside KC near the maturity/spawning ground was significantly negatively correlated with RPS. A significant negative correlation between the temperatures in winter and spring was detected in the area after 2000, when the conditions of the Pacific decadal oscillation index and the stability of the Kuroshio Extension were synchronous, indicating that KC shifted northward during this time. The reversed temperature pattern was consistent with the winter–spring movement of KC axis in the offshore direction and was correlated with the winter–spring difference in the intensity of the Aleutian low. These results suggest that the annual variation in chub mackerel recruitment after 2000 was strongly affected by the combined effects of ambient temperature because of the reversal of conditions that occurred between winter and spring around the maturity/spawning ground, which was related to the KC path.     

Ceramide aminoethylphosphonate from jumbo flying squid Dosidicus gigas attenuates the toxicity of cyanotoxin microcystin-LR

We have previously established the method for isolation of ceramide aminoethylphosphonate (CAEP) from jumbo flying squid Dosidicus gigas. In this study, we performed a MTT assay to evaluate the safety of CAEP to the cell lines for the application to health food and supplements. The CAEP did not show any cytotoxicity to various HEK293-transfectant cells. Next, we elucidated the positive function of CAEP to the somatic cells. Recently, we have reported that hepatotoxin microcystin-LR was taken up into the hepatocytes mediated by hepatocellular uptake transporters OATP1B1 and OATP1B3, and the cells were induced cytotoxicity subsequently. Cytotoxicity of microcystin-LR to permanently OATP1B3-expressing HEK293-OATP1B3 cells rather than to HEK293-OATP1B1 cells was preferentially attenuated by CAEP in a concentration-dependent manner. In addition, the enzyme activity of serine/threonine phosphatase, which was inhibited by microcystin-LR, was recuperated by co-exposure to CAEP. Furthermore, microcystin-LR-induced cellular protein phosphorylation were disrupted by CAEP exposure. These results suggested that CAEP is a promising remedy and/or preventive medicine for liver damage with microcystin-LR.