We have developed an enzyme-linked immunosorbent assay (ELISA) using two anti-Cry j I monoclonal antibodies (KW-S10 and KW-S131) for the quantitation of the major allergen from Japanese cedar (Cryptomeria Japonica) pollen, Cry j I. Polystyrene microplates coated with KW-S131 were incubated with pollen allergen extracts. Cry j I, which bound to the antibody, was detected with alkaline phosphatase-conjugated KW-S10 using chromogenic enzyme substrate. Cry j I could be measured in concentration of between 0.16 and 2.5 ng/ml by this assay. Intra- and inter-assay coefficients of variation were 1.1-3.5% and 0.9-4.6%, respectively. This assay was considered that it was specific for Cry j I of Japanese cedar pollen because it didn't react with allergens of hinoki (Chamaecypairs obtusa) pollen which have antigenicity in common to Japanese cedar pollen. This assay would be useful for the standardization of Japanese cedar pollen allergen extracts.
Lampteromyces japonicus (Japanese name, Tsukiyotake) is a poisonous and bioluminescent mushroom, which causes vomiting, stomachache and diarrhea.This mushroom is often mistaken by amateur mushroom hunters for the edible mushrooms Pleurotos osteatus (Hiratake), Lentinus edodes (Shiitake), and Panellus serotinus (Mukitake), since these species are all very similar in shape and color. Some people in the Shounai area of Yamagata Prefecture have eaten the salted poisonous mushrooms.We examined the gastrointestinal toxicity of the salted mushrooms and the nature of the toxic principles by using mice and frogs.Oral administration of the methanolic extract of L. japonicus to mice and frogs caused toxic effects such as swelling of the stomach in mice and vomiting in frogs. However, no toxic effects were associated with the extract of the salted mushroom.Isolation of the toxic principles from L. japonicus with guidance by biological assay was carried out by chromatographic separation, NMR and mass spectroscopic analysis revealed that illudin S was one of the toxic principles.
Ethyl acetate, n-butanol and water extracts of C. morifolium were examined for mutagenicity towards strains of Salmonella typhimurium (TA98 and TA100) with or without S9 mix. The ethyl acetate extract showed mutagenicity without S9 mix in TA98, but the mutagenicity was lost with S9 mix. The n-butanol and water extracts showed no mutagenicity in TA98 or TA100 with or without S9 mix.Antibacterial activities of the ethyl acetate extract were examined by using B. cereus, B. subtilis, M. luteus, St. aureus, E. coli, S. enteritidis, TA98 and TA100. This extract showed antibacterial activities towards gram-positive bacterial strains and TA98.
A serological typing scheme of Bacillus cereus has been developed by immunochemical analyses of flagellar antigen using an agglutination method. Enzyme-linked immunosorbent assay (ELISA) for the classification of flagellar serotype of Bacillus cereus had greater sensitivity. 10-500 times, than that of agglutination method. The specificity of flagellar antigen and antibody was determined by immunogold electron microscopy and ELISA inhibition assay. Application of ELISA is useful for the detection of the small amounts and many kinds of antigen-antibody reactions.
