Porphyromonas gingivalis has been implicated in adult periodontitis. A 75-kDa major outer membrane protein of P. gingivalis is an immunodominant antigen and is often associated with fimbrial proteins during its purification. The present study was performed to generate monoclonal antibodies (MAbs) to the 75-kDa protein and characterize the protein and the MAbs. Kight-week-old BALB/c mice were immunized with the partially purified 75-kDa protein from P. gingivalis 2561. Spleen cells of the mice were hybridized with SP2/0 Ag-14 mouse myeloma cells and then fused with polyethylene glyeol 3400. Hybrid cells surviving hypoxanthine/aminopterin/thymidine medium were tested for antibody secretion by ELISA. Positive clones were subcloned by limiting dilution and then three subclones were obtained. The subclones were injected peritoneally into Pristane-primed mice to produce MAbs. Immunoreactivity of the MAbs was confirmed by an immunoblot analysis. In the immunoblot using a partially denatured crude 75-kDa protein preparation as an antigen, the 75-kDa protein reacting with the MAbs revealed a ladder- like pattern which has been known as a characteristic of a native polymeric form of fimbriae. The intervals of the ladders, however, were different from those of fimbrial protein. The MAbs also recognized the completely denatured 75-kDa protein. Immunogold localization using IgG fraction of the MAbs demonstrated that the gold particles were deposited on globules on the surface of P. gingivalis 2561, but not on fimbriae. These results suggest that the 75-kDa protein is antigenically different from fimbrial protein and in nature, is not associated with, but may be physicochemically associated with the fimbrial protein during extraction and purification procedures for these proteins. Immunoreactivity of these 3 MAbs was tested by immunoblot using sonic extracts from various P. gingivalis strains. All the MAbs reacted only with the strains having proteins in their sonic extracts with a molecular mass of 75 kDa, but failed to recognize the strains having proteins with molecular masses of 61, 61.5, 76, and 78 kDa. These results indicate that strains of P. gingivalis have antigenically different major outer membrane proteins and the MAbs in the present study might have been raised against the same common, exposed immunodominant epitope in the 75-kDa proteins.
Animals ; Antibodies, Monoclonal* ; Chronic Periodontitis ; Clone Cells ; Enzyme-Linked Immunosorbent Assay ; Hybrid Cells ; Immunoglobulin G ; Membrane Proteins* ; Membranes* ; Mice ; Polyethylene ; Polymers ; Porphyromonas gingivalis* ; Porphyromonas* ; Spleen

No abstract available.

No abstract available.
Gonadotropin-Releasing Hormone* ; Humans

No abstract available.
17-alpha-Hydroxyprogesterone*

No abstract available.
Animals ; Female ; Ovary* ; Rats* ; Serine Proteases* ; Serine*

No abstract available.
Animals ; Female* ; Gonadotropin-Releasing Hormone* ; Haloperidol* ; Humans ; Rats*

No abstract available.
Heart Defects, Congenital*

No abstract available.
Child* ; Heart Arrest, Induced* ; Humans ; Oxygen*

No abstract available.
Fontan Procedure*

PURPOSE: Laparoscopic bladder neck suspension is an effective treatment for genuine stress urinary incontinence. To decrease the operating time, associated morbidity and hospital stay, we introduced laparoscopic bladder neck suspension using extracorporeal needle. We assessed the feasibility of this new technique, and compared with simple laparoscopic bladder neck suspension and transvaginal Raz procedure. MATERIALS AND METHODS: Eleven patients underwent a laparoscopic bladder neck suspension using extracorporeal needle were compared with 16 patients underwent simple laparoscopic bladder neck suspension and 66 patients treated with transvaginal Raz procedure for genuine stress urinary incontinence. All patients were assessed for their age, duration of symptom, operative time, length of hospital stay, postoperative residual urine volume, complications, and success rate. RESULTS: The result of present method was cure in 82% and improvement in 18% of patients. The overall success rate of three methods were similar. Average duration of postoperative admission was 3.4 days and average residual urine volume at discharge was 12.1 ml (simple laparoscopic Burch procedure: 5.1 days and 22.1 ml; Raz procedure: 7.7 days and 37.5 ml). Operative time of laparoscopic bladder neck suspension using extracorporeal needle was shorter than the simple laparoscopic bladder neck suspension and transvaginal Raz procedure (55 vs. 125 and 102 minutes). CONCLUSION: Laparoscopic bladder neck suspension using extracorporeal needle is a good technique for treatment of genuine stress urinary incontinence. However, additional long term follow-up study will be required.
Follow-Up Studies ; Humans ; Length of Stay ; Neck* ; Needles* ; Operative Time ; Urinary Bladder* ; Urinary Incontinence*