Biochemical analyses of the crustacean hyperglycemic hormone of the crayfish Astacus leptodactylus.

A biochemical analysis was made of the crustacean hyperglycemic hormone (CHH) of the crayfish Astacus leptodactylus, as present in the CHH-producing perikarya, in the axonal tract and in the sinus gland, respectively. Hyperglycemic material was analyzed by polyacrylamide gel electrophoresis (PAGE and SDS-PAGE) and high-pressure liquid chromatography (HPLC) in combination with a dotting immunobinding assay (DIA) and a bioassay for hyperglycemic activity. After electrophoretic analyses, the predominant biologically as well as immunologically detectable product present in all parts of the cell has an apparent molecular radius of approximately 7000 Da. In the perikarya extract, a second factor with lower electrophoretic mobility was found, which may represent the prohormone or precursor of CHH. The analyses by means of HPLC showed two predominant immunopositive peaks with an elution time of 28-29 and 52-54 min, respectively. For both HPLC peaks, electrophoretic analyses indicate a molecular weight of 7000 Da.

Adherence of oral treponemes and their effect on morphological damage and detachment of epithelial cells in vitro.

The present study shows adherence of Treponema denticola L12D and Treponema vincentii RitzA to guinea pig ear epithelial cells in vitro. The number of adhering organisms was positively related with the treponemal concentration and contact time. Incubation of washed T. denticola L12D and T. vincentii RitzA organisms or their culture supernatants with the epithelial cells induced morphological damage and detachment of these cells. In addition, indications for inhibition of epithelial cell proliferation were found. The activity of the supernatants was dose dependent, heat sensitive, and sensitive to sulfhydryl-containing components, suggesting the presence of enzymatic activity that might be of importance in the pathogenicity of these oral treponemes.

Effect of splint placement on black-pigmented Bacteroides and spirochetes in the dental plaque of beagle dogs.

The gingival microbiota in beagle dogs was investigated during a period of 42 days after the placement of continuous wire and arch bar splints. The splints were found to induce an increase in helicoidal-shaped micro-organisms and black-pigmented Bacteroides. Two different asaccharolytic Bacteroides types were isolated and tentatively identified as B. gingivalis and B. asaccharolyticus. In addition, B. intermedius-like organisms were isolated. The last two species were observed only after splint placement. The local accumulation of helicoidal-shaped micro-organisms in the epithelial tissue rather than in the micro-ulcerations was regarded as an indication of an active role for these spirochetes in the disease process. The observed shift toward tissue-invading and potentially pathogenic micro-organisms after splinting stresses the need for antimicrobial measures during splint placement in order to minimize periodontal destruction.