Rapid and simple determination of histamine-N-methyl transferase activity by high-performance liquid chromatography with UV detection.

A rapid, simple and low-cost assay method of histamine-N-methyltransferase activity was developed. Methylhistamine, which was separated from the enzymatic reaction system on reversed-phase high-performance liquid chromatography using an ion-paired chromatographic technique, was detected spectrophotometrically at 226 nm. The mobile phase used for the separation of methylhistamine was 0.05M NH4H2PO4 (pH 3.0) containing 2 mM of sodium octanesulfonate. The new assay technique could detect methylhistamine as an enzyme activity product of histamine-N-methyltransferase in the brain and kidney of rats. Chloropheniramine maleate, an antihistamine, activated the histamine-N-methyltransferase. Whether neurotransmitter or neuromodulator, the role of histamine in the brain has not yet been made clear. Therefore, the present method could be applicable for the enzymatic investigation of histamine metabolism in central nervous system or inflammatory reactions.

Analysis of defense mechanisms against bacterial infection by oral streptococcus in normal flora.

The incidence of oral alpha-streptococci with inhibitory activity against pathogens as a defense mechanism in the oral cavity was investigated in healthy individuals. Inhibitory strains were isolated from tonsil, tongue, cheek, saliva and dental plaque, and the detection rate of these strains isolated from tonsil was the highest. These results suggested that tonsillar flora is most important as a defense mechanism of the oral cavity. With respect to the effects of antibiotics against inhibitory alpha-streptococci, minimal inhibitory concentration of 90% of cells (MIC90) of PCG, ABPC, CCL, CFIX and EM against inhibitory alpha-streptococci, and relative detection rates of inhibitory alpha-streptococci before and after antimicrobial therapy were investigated. MIC90s of all antibiotics against these strains were low and sensitive to antibiotics tested. However, in vivo, detection rates of these strains before and after therapy did not differ significantly. Therefore, inhibitory strains were not affected by antibiotics as their MIC90 were low during short term medication.

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.