MMP-9 activity in urine from patients with various tumors, as measured by a novel MMP activity assay using modified urokinase as a substrate.

Matrix metalloproteinases (MMPs) play an important role in many pathologic processes, but their activities are difficult to determine since no simple specific and/or chromogenic substrate exists. We have developed a novel MMP activity assay using a modified urokinase as a substrate. Protein engineering enabled the plasmin activation site in this urokinase to be substituted by a specific activation site recognized by MMPs. In this way the MMP activity can be monitored via urokinase activity as measured by a simple chromogenic assay. The assay was made specific for MMP-9 by a capture step using MMP-9-specific antibodies that do not interfere with MMP-activity. This assay monitors the amount of active enzyme as well as the latent, but potentially active proform. Using this assay the levels of MMP-9 were investigated in urine from patients with various kinds of carcinoma. High levels of both active and latent MMP-9 were detected in urine from patients with carcinoma of the bladder, whereas hardly any activity was observed in urine from healthy controls. MMP-9 in urine was present in its intact form. Surprisingly, MMP-9 was also increased in the urine of patients with nonurogenital carcinoma. Therefore, measurement of urinary MMP-9 activity levels may be a convenient diagnostic tool for various types of carcinoma.

Epidemiology and transmission of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans among children and their family members. A report of 4 surveys.

The distribution and transmission of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in 4 families were studied. The families were included, based on the isolation of P. gingivalis from a young child or adolescent. The probands of these 4 families were: a 5-year old periodontally healthy boy; a 17-year old girl with severe generalized juvenile periodontitis; an 11-year old girl with prepubertal periodontitis; 2 sisters, 5 and 17-years old, with untreated severe periodontitis as a component of the Papillon-Lefèvre syndrome. All members of the 4 families were examined clinically and microbiologically for the presence of P. gingivalis and A. actinomycetemcomitans. Most of the parents appeared to be adult periodontitis patients; the parents of one proband were edentulous. Results showed that in all cases at least one of the parents was positive for P. gingivalis. On the basis of indistinguishable restriction endonuclease patterns (REPs) of P. gingivalis and A. actinomycetemcomitans isolates from parents and their children, and distinct REPs from unrelated individuals, the present study indicates that P. gingivalis and A. actinomycetemcomitans were transmitted between parents and their children.

Transmission of Porphyromonas gingivalis between spouses.

Porphyromonas gingivalis has been associated with severe forms of periodontitis. The question can be raised about the origin of this suspected periodontal pathogen. The purpose of the present investigation was to study the possibility of transmission of P. gingivalis between spouses. 18 patients were selected with severe periodontitis and colonized with P. gingivalis. 10 of their spouses appeared to be colonized with P. gingivalis too. 8 of the patients and their spouses were investigated further clinically and microbiologically. Microbiological evaluation revealed mostly high %s of P. gingivalis in the pockets, not only of the patients (5-48% of the cultivable flora), but also in 7 of the 8 spouses (0.2-61%). Furthermore, this species was isolated often from the saliva, the tongue, the buccal mucosa and the tonsillar area from both patients and spouses. For typing purposes, bacterial DNA was isolated, treated with the restriction endonucleases Bam HI or Pst I, after which the DNA fragments were separated by agarose gelelectrophoresis. With one exception, each individual was colonized with only one clonal type of P. gingivalis. The DNA patterns of all P. gingivalis isolates from unrelated individuals were found to be distinct. In contrast, in 6 of the 8 couples, the DNA patterns of isolates from husband and wife were indistinguishable. From these data, it can be concluded that it is most likely, that P. gingivalis can be transmitted between spouses. It remains to be investigated whether transmission of P. gingivalis is a risk factor for developing periodontal destruction.