Molecular basis of salivary proline-rich protein and peptide synthesis: cell-free translations and processing of human and macaque statherin mRNAs and partial amino acid sequence of their signal peptides

Acidic proline-rich phosphoproteins and phosphopeptides are abundant components of parotid and submandibular salivary secretions in man and in the subhuman primate, Macaca fascicularis. The major acidic proline-rich proteins and the proline-rich phosphopeptide, statherin, of man and macaques have been shown to be potent inhibitors of calcium phosphate precipitation and are thought to function in the oral environment by maintaining saliva supersaturated with respect to calcium phosphate salts. Little is known about the biosynthesis of these proline-rich phosphoproteins and peptides, and the aim of the present work was to determine the structural relationship between statherin precursors and native human and macaque statherin. RNA was isolated from human submandibular gland, and poly(A+) mRNA was selected by affinity chromatography on oligo(dT) cellulose and translated in a reticulocyte lysate. Electrophoretic analysis of the translation products revealed that this mRNA directed the synthesis of a large number of polypeptides with Mrs ranging from 5000 to 70,000. Immunoprecipitates, prepared with an antiserum directed against human statherin, contained a single component with a Mr of 7800, approximately 2000 daltons larger than native statherin. Radiosequencing of the in vitro precursor of statherin in immunoprecipitates demonstrated the presence of a 19-residue signal peptide. These results suggest that statherin is derived from a unique structural gene, and does not result from proteolytic processing of a large polyprotein precursor.     

Colonization of the human oral cavity by a strain of Streptococcus mutans

Streptococcus mutans strain JH1001 produces a bacteriocin that can kill virtually all other strains of this micro-organism. The ability of JH1001 to colonize the human oral cavity was tested in a study involving five subjects and three different infection regimens, all of which involved multiple exposures to large numbers of organisms. Two and one-half years after infection, JH1001 was found to have persistently colonized three of the subjects. The indigenous S. mutans in one subject were reduced below the level of detection. Levels of (total) S. mutans and S. sanguis were not affected in persistently colonized subjects. Mutants of indigenous S. mutans resistant to the bacteriocin were not observed. The results indicate the importance of host variability and infection regimen for superinfection by this strain of S. mutans. The efficient replacement of indigenous S. mutans by JH1001 in one subject lends support to the eventual application of replacement therapy to the prevention of dental caries.     

Cytotoxicity of Epiphany and Resilon with a root model

AIM: To record the cytotoxicity of Resilon and Epiphany (Pentron clinical technologies, Wallingford, CT, USA) using a root model. METHODOLOGY: Thirty teeth with single roots were sectioned at the enamel-cementum junction, the root canals prepared and each root then sterilized before filling with the lateral condensation technique using one of three filling materials (n = 10 per group): Resilon and Epiphany, Sealite (Septodont, Pierre Rolland, Merignac, France) and gutta-percha, Roekoseal Automix (Coltène/Whaledent, Langenau, Germany) and gutta-percha. The roots were stored at 37 degrees C in an incubator to allow for setting of the root filling materials. The apices of the roots were dipped in 1 mL of MEM culture medium for 1, 2, 7 and 30 days renewing the medium every day. After 24 h contact between the medium and the filled roots, the medium was used to measure the cytotoxicity on mouse fibroblasts L 929 with the MTT assay that recorded the mitochondrial activity of the target cells. An additional test according to ISO 10993-5 standards was undertaken to compare Resilon and Epiphany. RESULTS: The root model showed no statistically significant differences between the sealers at 7 and 30 days (NS). Epiphany and Resilon were the most cytotoxic materials at 1 and 2 days (P < 0.001). Unlike Epiphany, Resilon was not cytotoxic when tested according to ISO 10993-5 standards. CONCLUSIONS: The cytotoxicity of Resilon + Epiphany, due mainly to Epiphany, decreased after 2 days to reach a level comparable with commonly used root canal sealers.     

Rectal ketamine and midazolam for premedication in pediatric dentistry

Rectally administered midazolam (0.30 mg/kg) and ketamine (5 mg/kg) were compared for preanesthetic medication in children undergoing dental extractions. Sixty patients between the ages 2 and 9 years were randomly allocated to three groups in this double-blind study. In one group of patients who received ketamine rectally, intravenous midazolam (0.05 mg/kg) also was administered immediately after induction of anesthesia. The results from this trial show that 30 minutes after rectal administration of the two drugs, good anxiolysis, sedation, and cooperation were obtained in most patients. Although midazolam appeared to be marginally more efficacious than ketamine in the majority of assessments made and seemed to have less adverse effects, no statistically significant differences could be shown. Ketamine showed a slight decrease and midazolam a slight increase in average blood pressures after premedication. These blood pressure differences were, however, considered to be of little clinical importance.     

Effects of different surgical regimes on nasal asymmetry and facial attractiveness in patients with clefts of the lip and palate

Surgery has often been blamed for the subsequent poor growth and deformity of the nasal complex in patients with cleft lip and palate. This study compares two groups of patients with unilateral clefts of the lip and palate for the effect that different surgical techniques have on nasal symmetry and facial attractiveness.