Detection of Bacillus cereus flagellar antigen by enzyme-linked immunosorbent assay (ELISA)

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.     

Synthesis of human gamma-glutamyl transpeptidase (GGT) during the fetal development of liver

We have determined expression of human GGT gene encoding gamma-glutamyl transpeptidase (GGT) during fetal development of liver using the Northern-blot analysis with a cloned human GGT cDNA and immunohistochemistry with a monoclonal antibody. GGT mRNA could be detected as early as the 12th week of gestation. It then increased gradually to a peak of approx. threefold the amount at week 12, at week 40, just before birth. The size of the mRNA in the fetal liver was 2.7 kb and mRNA of the same size was detected both in the human fetal kidney and human hepatocellular carcinoma as well as normal adult liver. Immunohistochemical analyses show that GGT increased as the fetal liver developed in parallel with the increase in mRNA. Histochemically, GGT was shown to be located in the wall of bile canaliculi when synthesis was low in early development, but to be distributed, in addition, all over the cell membrane of the fetal hepatocytes when synthesis was high at the later stage of development.     

Amplification and substantial purification of cardiolipin synthase of Escherichia coli.

A simple, specific, and sensitive assay procedure for cardiolipin synthase of Escherichia coli has been developed. This measures the radioactivity of glycerol formed from phosphatidyl [2-3H]glycerol and is mainly based on the findings that 400 mM phosphate and 0.015% Triton X-100 markedly activate the enzyme. Cardiolipin synthase was amplified 760-fold upon induction with isopropyl beta-D-thiogalactoside in cells harboring a pBR322 derivative in which the cls gene encoding this enzyme was preceded by the tac promoter. Under these conditions, cardiolipin content increased, membrane potential decreased, spheroplasts became fragile, cells lost viability, and inducer-resistant mutants appeared at a high frequency. The amplification enabled the isolation of an enzyme preparation with a specific activity approximately 10,000-times higher than that of wild-type whole cell lysate. This purification was simply achieved by extraction of the crude membrane fraction with Triton X-100 and a single phosphocellulose column chromatography. This preparation, together with the crude envelope fraction, was used to characterize the basic properties of E. coli cardiolipin synthase, some of which were utilized in setting up the assay conditions.     

In vitro propagation ofAtractylodes lancea

Shoot cultures ofAtractylodes lancea DC. (Compositae) have been established by inoculating the flower bud on Linsmaier-Skoog's medium supplemented with 10⁻⁵ M naphthaleneacetic acid and 10⁻⁵ M 6-benzyl-aminopurine. Shoots were multiplied on a medium containing 10⁻⁶ M 6-benzylaminopurine. Propagated shoots rooted on a medium devoid of any plant growth regulators were transferred to potting soil and finally to the field.     

The mechanism of dNTP-unbalanced cell death induced by 5-fluorouracil and its derivatives

The mechanism of 5-fluorouracil (5-FU) and 5-fluorodeoxyuridine (FdUR)-induced death of mouse mammary tumor FM3A cells was studied. When the cells were exposed to 5-FU or FdUR, an unbalance of intracellular dNTP pool resulted. The unbalance was followed by breakage of mature DNA. DNA double strand breaks were observed in the FdUR (1 microM) treated cells 16 hrs after the administration. We assume that the double strand breaks play an important role in the mechanism of the FdUR-mediated cell death. In addition, the activity that can induce DNA double strand breaks was detected in the lysate of FdUR treated FM3A cells. Since intracellular dNTP pool unbalance seems to be the trigger of these events, this phenomenon may be termed as dNTP-unbalanced cell death.     

The large-scale detoured migration route and the shifting pattern of migration in Oriental honey-buzzards breeding in Japan

We describe the detoured migration route of the Oriental honey-buzzard Pernis ptilorhyncus , showing differences between autumn and spring migration, using data from 10 adult individuals marked with satellite transmitters. In autumn, the migration routes were very similar from Japan to the south end of the Malay Peninsula. The wintering sites were distributed within the Philippines, Borneo and the Malay Archipelago. During autumn, migration of the birds had few long-term stopover sites, instead, sometimes decidedly slowing their migration rate while proceeding in a consistent direction. During spring migration, the honey-buzzards penetrated into southern China, moving north to the base of the Korean Peninsula. The birds then went south through the Korean Peninsula to reach Japan. Before travelling to China, all spring migrants stopped for several weeks in south-east Asia. The slow rate of travel in the autumn suggests that migrants were foraging and replenishing their energy reserves. Instead of a migration strategy that uses only a few long-term stopover sites, honey-buzzards may adopt a strategy based on a number of short-term stay sites.