Anti type I allergic property of Japanese butterbur extract and its mast cell degranulation inhibitory ingredients

Pollenosis is a disease that affects 1 in 10 of the Japanese population. During the season of cedar pollen dispersal, many patients suffer from symptoms such as sniffling, sternutation, and itching of the eyes. Japanese butterbur is a popular vegetable and is one of the few domestic vegetables in Japan. The anti type I allergic effects of an aqueous ethanol extract from aerial parts of Japanese butterbur (JBE) were evaluated in rats and RBL-2H3 mast cells. In the passive cutaneous anaphylaxis reaction in rats, a single oral treatment of JBE (1000 mg/kg) was found to suppress the reaction. In IgE-sensitized RBL-2H3 cells, JBE (10-100 microg/mL) inhibited beta-hexosaminidase release, leukotriene C(4)/D(4)/E(4) synthesis, and TNF-alpha production. Moreover, a high concentration of JBE (1000 microg/mL) suppressed smooth muscle constriction induced by histamine (10 microM) and leukotriene D(4) (10 nM) in a guinea pig trachea strip. The search for components in JBE with an inhibitory activity on mast cell degranulation was guided by inhibition of beta-hexsosaminidase release. Two eremophilane-type sesquiterpenes, six polyphenolic compounds, and two triterpene glycosides were isolated. Of these compounds, fukinolic acid, a principal polyphenol constituent, showed potent inhibitory activity (IC(50) value = 2.1 microg/mL). Consequently, On the basis of its inhibition of mast cell activation and direct smooth muscle reaction induced by released mediators, JBE was found to suppress the type I allergic reaction.     

Acid Dissolution of Olivines, Feldspars and Dunite

We have treated feldspars (orthoclase, albite, anorthite), JF-1 (a mixture of orthoclase and albite), olivines (fayalite, forsterite) and JP-1 (dunite: an olivine-rich rock) with HNO₃ solution at pH 2 for 10 – 1200 min. The resultant changes in surface composition were assessed by X-ray photoelectron spectroscopy (XPS). The XPS data for fayalite, notably shifts in the Si 2s and O 1s binding energies, indicate preferential dissolution of Fe and formation of a Si-rich surface layer composed of amorphous silica. A Si-rich layer also forms on the surface of the other minerals as the result of acid treatment but the changes in binding energies are relatively small. Forsterite and anorthite dissolved almost congruently in acid solutions, and the Si-rich surface layer is poorly developed. Unlike forsterite, JP-1 shows preferential leaching of Mg relative to Si. Although forsterite is the major constituent of JP-1, this rock also contains some talc and orthopyroxene. These minor mineral constituents appear to influence the behavior of JP-1 in acid solutions. On the other hand, the dissolution and resultant surface alteration of JF-1 were comparable to those of its constituent minerals. The extent of Si-rich surface layer formation followed the order of albite = orthoclase > anorthite for the feldspars, and fayalite > forsterite for the olivines.     

Effect of stocking density and addition of proteins on larval survival in Japanese flounder, Paralichthys olivaceus

The effects of larval density, rearing media, and protein additions on fish larval survival were tested by rearing larvae in small containers. Yolk-sac larvae of Japanese flounder, Paralichthys olivaceus, were transferred 1 day after hatching to plastic dishes and cell-culture microplates by Pasture pipette at densities ranging from 0.3 to 30 individuals/ml and incubated at 16 °C without food. Dead larvae were removed and counted every 12 h. Three peaks of larval mortality were observed: the first peak appeared just after the transfer, the second corresponded to the completion of yolk absorption, and the third occurred 10–12 days after the transfer. Since about 50% of larvae survived until the third peak, it was clear that mortality at the second peak (completion of yolk absorption; equivalent to the first feeding) was not due to starvation. At the first mortality peak, better survival was unexpectedly recorded at higher larval densities (30 individuals/ml), suggesting that some material beneficial for survival is derived from the larvae. This idea was further supported by the improved survival in media previously used to rear larvae at high densities. The addition of bovine serum albumin (BSA) at 10 μg/ml effectively reduced mortality in microplates and in 30-l tanks. To reduce the larval mortality just after handling, addition of egg white at a dilution rate of 1/30,000 (vol/vol) or less is recommended as a practical alternative to BSA. Since the addition of polyethylene glycol 6000 (1 μg/ml) also effectively reduced mortality, reduction in mortality could not be attributed to a specific mechanism. The precise determination of the magnitude and timing of larval mortality was attained through the use of small containers, and a method to prevent mortality due to handling was identified in this study. These types of tests may allow the logical evaluation and establishment of better rearing procedures for seed production in aquaculture.     

Lipid characteristics of coastal migratory Sarda orientalis tissues

To characterize the 22:6 n-3 (docosahexaenoic acid, DHA) levels of the oriental bonito Sarda orientalis, a coastal migratory tuna species, total lipids (TL) of white muscle, dark muscle, liver, pyloric cecum, gonad, and other viscera were separated into lipid classes, the constituents of TL were quantified, and the fatty acid composition of TL, triacylglycerols (TAG), phosphatidylethanolamine, and phosphatidylcholine was analyzed. The crude lipid contents of muscle and other organs were 0.8–3.2% and 2.3–9.1%, respectively. DHA was found in TL of various organs at levels ranging from 19.2% to 27.6% in muscle and 16.3% to 28.5% in other organs. The levels of DHA in muscle TAG (8.2–16.0%) were lower than or comparable to those in visceral TAG (6.9–24.0%). These findings did not coincide with those observed in active-migratory tuna species, which accumulate DHA in their muscle TAG during migration. These findings suggest that the DHA distribution of S. orientalis is different from that observed in active-migratory tuna species, and that the differences may be due to migration type of the fish.     

Supplemental effect of bile salts to soybean meal-based diet on growth and feed utilization of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

A feeding experiment was conducted to evaluate the supplemental effect of bile salts to a defatted soybean meal-based non-fish meal diet for rainbow trout Oncorhynchus mykiss. A fish meal-based diet (FM) and two non-fish meal diets with and without bovine bile salts (SC+B and SC, respectively) were fed to fish (13 g initial weight) for 10 weeks. Fish fed diet SC showed inferior growth and feed efficiency, while bile salt supplementation improved the parameters to the same levels as fish fed diet FM. Crude fat and starch digestibility of diet SC-fed fish decreased after the 10-week feeding trial compared to the data obtained with fish that had no experience of the diet. Total biliary bile salt content and intestinal maltase activity of fish fed diet SC were the lowest among treatments, while these parameters were improved by bile salt supplementation. Morphological changes occurred in the distal intestine and liver of the diet SC group, although the histological features of fish fed diet SC+B were similar to those of fish fed diet FM. These results suggest that bile salt supplementation to a soybean meal-based diet improves the nutrient utilization by normalizing digestive processes in rainbow trout.     

Preparation of β-1,3-glucanase from scallop mid-gut gland drips and its use for production of novel heterooligosaccharides

The mid-gut gland of scallop Patinopecten yessoensis has been discarded in scallop processing factories as a fishery waste and various attempts have been made to turn the waste into valuable resources. In the present study, we tried to use mid-gut gland drips from scallop as a source of β-1,3-glucanase. The mid-gut gland drips were collected in a local fishery factory in Yubetsu-cho, Hokkaido Prefecture. β-1,3-Glucanase was purified from the mid-gut gland drips by ammonium sulfate fractionation followed by successive chromatography on Toyopearl Phenyl-650M and Toyopearl DEAE-650M. The scallop β-1,3-glucanase, named PyLam38 in the present study, showed a molecular mass of approximately 38 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis, and hydrolyzed laminarin, a β-1,3-glucan from Laminaria sp., producing laminaribiose and glucose with an optimal pH and temperature of 6.0 and 45°C, respectively. PyLam38 exhibited high transglycosylation activity toward various accepter substrates such as monosaccharides, alcohols and xylooligosaccharides. Thus, PyLam38 was found to be useful for the production of various novel heterooligosaccharides consisting of laminarioligosaccharides and various accepters.     

The Colletotrichum higginsianum secreted effector protein ChEC91 induces plant cell death

ChEC91, a novel cell death-inducing effector protein from the fungal pathogen Colletotrichum higginsianum, causal agent of crucifer anthracnose disease, is described. Both transient expression of ChEC91 and infiltration of purified recombinant protein induced necrotic lesions in Nicotiana benthamiana leaves. The recombinant protein also induced electrolyte leakage and callose deposition in Arabidopsis thaliana leaf tissue and the expression of defence marker genes. Moreover, fungal mutants constitutively over-expressing ChEC91 in C. higginsianum were impaired in appressorial penetration on Brassica rapa cotyledons. These results suggest that inappropriate expression of ChEC91 might negatively affect the early stage of C. higginsianum infection by inducing plant defence responses. Protein domain deletion analysis showed that the C-terminal region of ChEC91 was necessary, but not sufficient, for activity in N. benthamiana. Homologous effector proteins cloned from C. gloeosporioides, Fusarium graminearum, and Pyricularia oryzae differed in their cell death-inducing activity, which appeared related to sequence variations in the C-terminal region of these proteins. Moreover, this region contained amino acid residues that were well conserved within Colletotrichum species. These results suggest that the amino acid residues in the C-terminal region may be important for inducing cell death in plants.