Eiji Inoshita

Inhibitory Effect of Human Plasma and Saliva on Co-aggregation Between Bacteroides gingivalis and Streptococcus mitis

The effect of human plasma and saliva on co-aggregation between Bacteroides gingivalis and Streptococcus mitis was studied by means of a turbidimetric assay. The co-aggregation activity was obtained from the maximum slope of the absorbance vs. time curve. Its dependence on pH, temperature, and ionic strength was examined, and the number of Bacteroides cells in relation to the number of Streptococcus cells resulting in optimal co-aggregation was established. Co-aggregation inhibition experiments showed that the co-aggregation activity was inhibited by 1-arginine and 1-lysine, although the activity was unaffected by the sugars tested. Human plasma and saliva were able to inhibit the co-aggregation in a dose-dependent reaction. Plasma exhibited the most potent inhibitory activity in these fluids. Fibrinogen was the most potent inhibitor of the plasma-derived proteins tested. These data suggest the possibility that the oral fluids may modulate the attachment of B. gingivalis to Gram-positive bacteria in periodontal pockets.

Effect of Transferrin on the Growth of Porphyromonas gingivalis

This study describes the effect of transferrin as an iron source on the growth of Porphyromonas (formally Bacteroides) gingivalis. Bacterial growth was monitored spectrophotometrically. All strains of P. gingivalis tested grew well in medium containing transferrin. The growth of P. gingivalis depended not only on the concentration of transferrin, but also on the iron saturation level of the protein. However, growth was not stimulated with either the ferrous or ferric iron salts tested. The addition of dipyridyl to the medium containing transferrin suppressed the growth of P. gingivalis, which also did not show species-specificity for human transferrin. Transferrin-binding activity was found in P. gingivalis by solid-phase assay with peroxidase-conjugated human transferrin. These results suggest that P. gingivalis may be capable of utilizing transferrin as an iron source for growth in vivo.

Changes in Oxygen Consumption in Dog Gingiva during Induction of Experimental Periodontitis

The purpose of this study was to examine changes in oxygen consumption in dog gingiva during induction of experimental periodontitis. The disease was induced in adult mongrel dogs during a 16-week period by placement of silk ligatures around selected teeth. The oxygen consumption rate of gingival tissue was determined in vivo by a non-invasive technique, tissue reflectance spectrophotometry. Changes in such clinical parameters as gingival index, plaque index, pocket depth, attachment level, and gingival crevicular fluid flow indicated acute inflammatory responses during the first three weeks after ligation, followed by the appearance of chronic inflammation during the remaining 13 weeks. The oxygen consumption rate increased during the first seven days after ligation and stayed near the maximum level for 2-7 weeks; this was followed by a gradual decrease during the final nine weeks. These results suggest that gingival oxygen consumption increases rapidly with the increase of acute inflammation responses and then decreases slightly with the gradual development of chronic inflammation. Positive correlations were observed between the oxygen consumption rate and other clinical indices. Thus, the tissue reflectance spectrophotometry is a new, useful method for objective, quantitative, and non-invasive assessment of gingival oxygen consumption.

Purification and Some Properties of Fucosyltransferase in Human Parotid Saliva

Fucosyltransferase was purified from human parotid saliva by affinity chromatography on GDP-hexanolamine Sepharose, followed by chromatofocusing on PBE 94 exchanger gel. The purified enzyme had the N-acetylglucosaminide α1→4, the N-acetylglucosaminide α1→3, and the glucoside α1→3 fucosyltransferase activities. The molecular weight of the purified enzyme was estimated to be approximately 20,000. These enzyme activities showed identical pH and divalent metal ion dependencies and identical rates of inactivation upon being heated. The paper chromatographic analysis of the fucosylated products by the purified enzyme and the susceptibility of these products to linkage-specific fucosidase digestion indicated that the transferase formed the Fuc α1→4GlcNAc, Fuc α1→3GlcNAc, and Fuc α1→3Glc linkages.

The Presence of Fucosyltransferases with Different Substrate Specificity in Human Parotid Saliva

Using glycoproteins and milk oligosaccharides as substrate acceptors, we demonstrated at leaet two fucosyltransferases in human parotid saliva. One enzyme transferred L-fucose from GDP-fucose to the C-3 position of N-acetylglucosamine or glucose residue of oligosaccharide chains, and the other transferred to the C-4 position of N-acetylglucosamine residue of oligosaccharide chains.