A food‐borne outbreak of gastroenteritis due to genotype G1P[8] rotavirus among adolescents in Japan

Six high school students in Tochigi prefecture, Japan, developed gastroenteritis after eating at a pork cutlet shop. Molecular epidemiologic analyses showed that the causative agent was genotype G1P[8] rotavirus (RV), this being detected in stool samples from both the patients and the asymptomatic food handlers. The detected RV strains were closely related genetically. The only uncooked food that all victims had eaten was raw sliced cabbage. These findings results suggest that uncooked foods contaminated with RV may be sources of infectious gastroenteritis in adolescents.     

Binding sites of an aminoglycoside in the cochlea examined by immunocytochemistry

To localize the binding sites of aminoglycosides in the cochlea, immunocytochemistry was used with the antibody to gentamicin and the protein-A/gold complex. We found that the main binding sites were the stereocilia, the cuticular plates of hair cells, the head plates of Deiters' cells, cell filaments and the cones of pillar cells, tectorial membranes, basilar membranes, the matrix of the spiral limbus, plasma membranes, mitochondria, and the chromatin of various kinds of cells. Triphosphoinositide and acidic glycosaminoglycans are the two most likely candidates for the cause of binding activity.     

Detection of soluble interleukin-2 receptor and soluble intercellular adhesion molecule-1 in the effusion of otitis media with effusion

We measured sIL-2R, TNF-alpha and sICAM-1 in the sera and middle ear effusions (MEEs) of patients with otitis media with effusion (OME). Although there was no signmcant difference between the sIL-2R levels of the serous and mucoid MEEs, they were significantly higher than serum sIL-2R levels of OME patients and healthy controls. TNF-alpha levels of the mucoid MEEs were significantly higher than those of the serous type. However, TNF-alpha was rarely detected in the sera of OME patients or healthy controls. We observed significant differences between the serous and mucoid MEEs with respect to their sICAM-1 levels, which were also higher than serum slCAM-1 levels of OME patients and healthy controls. Our findings suggested that IL-2, TNF-alpha and ICAM-1 could be significantly involved in the pathogenesis of OME through the cytokine network.     

Polymyxin-B-binding sites in the cochlea as demonstrated by polymyxin-B/gold labeling

Summary A polymyxin-B/bovine-serum-albumin/gold complex was used as a probe to detect the binding sites of polymyxin B on thin sections of cochlea embedded in Spurr's resin. The binding sites were found to be mainly located on the stereocilia, the cuticular plate of hair cells, the head plate of Deiters' cells, the tonofilaments in pillar cells and Deiters' cells, fibrous structures in the spiral limbus, the tectorial membrane and the basilar membrane and neural elements such as nerve endings, fibers, and the myelin sheath. The mitochondria, plasma mimbrane, and chromatin of the nuclei of the cells observed also exhibited binding. Our results suggest that phospholipids, glycoconjugates, cytoskeletal proteins and nucleic acids are responsible for this binding activity.     

Chemical modification of the sugar part of methyl acarviosin: synthesis and inhibitory activities of nine analogues.

Nine analogues of methyl acarviosin (1), the core structure of acarbose and its homologues, the 6-hydroxy-(2), 6-azido-(3), 6-amino- (4), 6-acetamido-(5), 6-methoxy-(6), 6-hydroxy-2-O-methyl-(8), and 6-hydroxy-3-O-methyl derivatives (9), including the 5-methoxycarbonyl analogue (7) and 3,6-anhydro derivative (10) of 2, were synthesized by chemical modification of the sugar part of 2 derived by condensation of methyl 3,4-anhydro-alpha-D-galactopyranoside (17) and 4,7:5,6-di-O-isopropylidenevalienamine (26) or by direct coupling between 26 and the 6-substituted methyl 3,4-anhydro-alpha-D-galactopyranoside derivatives. Compounds 2 and 8 show notable inhibitory activity against yeast alpha-D-glucosidase almost comparable to that of 1. Introduction of a polar substituent at C-6 of 1 decreases the inhibitory activity. Interestingly, inversion of the conformation of the sugar part of 1 by introduction of the 3,6-anhydro bridge elicits almost no effect on the inhibitory activity.     

Rapid Preparation of Stable Isotope Labeled Peptides that Bind to Target Proteins by a Phage Library System

We have developed a system for directly isolating foreign peptides displayed on the N-terminus of the major coat protein of bacteriophage M13. The phage particle in this system is formed as a mixture of wild type and modified coat proteins. The N-terminal segment of the modified coat protein was mutated for chemical cleavage, in order to obtain the displayed peptide from the major coat protein. Using ¹³C, ¹⁵N- labeled medium, we introduced stable isotopes, ¹³C and/or ¹⁵N, into the coat proteins. The NMR spectra for the cleaved peptides from the phage particles could be recorded within a few days after the selection of the phage clone.     

Characterization of DNA recognition by the human UV-damaged DNA-binding protein.

The UV-damaged DNA-binding (UV-DDB) protein is the major factor that binds DNA containing damage caused by UV radiation in mammalian cells. We have investigated the DNA recognition by this protein in vitro, using synthetic oligonucleotide duplexes and the protein purified from a HeLa cell extract. When a 32P-labeled 30-mer duplex containing the (6-4) photoproduct at a single site was used as a probe, only a single complex was detected in an electrophoretic mobility shift assay. It was demonstrated by Western blotting that both of the subunits (p48 and p127) were present in this complex. Electrophoretic mobility shift assays using various duplexes showed that the UV-DDB protein formed a specific, high affinity complex with the duplex containing an abasic site analog, in addition to the (6-4) photoproduct. By circular permutation analyses, these DNA duplexes were found to be bent at angles of 54 degrees and 57 degrees in the complexes with this protein. From the previously reported NMR studies and the fluorescence resonance energy transfer experiments in the present study, it can be concluded that the UV-DDB protein binds DNA that can be bent easily at the above angle.