bluuebunny/crossref_metadata_2025 huggingface dataset:
Research Reports Clinical
Authors: Seow W.K. , Wan A.
Published: January 2009

The aim of the present investigation was to compare the dimensions of the primary incisors from pre-term children and full-term controls. One hundred and eleven pre-term children, consisting of 86 very-low-birthweight (< 1500 g), 25 low-birthweight (from 1500 to 2500 g), and 169 full-term, normal-birthweight (> 2500 g) children, donated a total of 572 maxillary and mandibular primary central and lateral incisors for study. The teeth were measured by means of a digital micrometer. The results showed that there was a dose-response effect of birthweight on tooth size. The very-low-birthweight teeth showed the smallest dimensions, the normal-birthweight controls the largest, and the low-birthweight teeth intermediate dimensions (p < 0.001). In the maxillary primary central and lateral incisors, and the mandibular primary central incisors, very-low-birthweight teeth were from 6 to 11% smaller in both mesiodistal and faciolingual dimensions compared with normal-birthweight teeth (p < 0.001). The largest differences were observed in the maxillary lateral incisors, where mean decreases of 0.58 ± 0.45 mm in mesiodistal and 0.50 ± 0.40 mm in faciolingual dimensions (11% reduction in both measurements) were observed. In addition, in the mandibular and maxillary lateral incisors of very-low-birthweight children, the left-sided teeth were significantly smaller than those on the right side in both mesiodistal and faciolingual dimensions (p < 0.03).
Localization of EMSP1 Expression During Tooth Formation and Cloning of Mouse cDNA
Authors: Hu J.C-C. , Ryu O.H. , Chen J.J. , Uchida T. , Wakida K. , Murakami C. , Jiang H. , Qian Q. , Zhang C. , Cttmers V. , Bartlett J.D. , Simmer J.P.
Published: January 2009

Enamel Matrix Serine Proteinase 1 (EMSP1) is a proteolytic enzyme that has been isolated from the developing enamel of pig teeth. Its apparent function is to degrade the organic matrix in preparation for enamel maturation. The expression of EMSP1 has never been investigated in another organism besides the pig, and EMSP1 expression in the enamel organ has never been specifically demonstrated in ameloblasts. Here we report the expression of recombinant pig EMSP1 (rpEMSP1), the generation of rabbit polyclonal antibodies against rpEMSPl, the characterization of the antibodies and EMSP1 expression by Western blot and immunohistochemical analyses, the cloning and characterization of a full-length cDNA encoding mouse EMSP1, and the localization of EMSP1 expression in ameloblasts in mouse day 14 first and second molars by in situ hybridization. The full-length mouse EMSP1 cDNA clone has 1237 nucleotides, excluding the poly(A+) tail, and encodes a preproprotein of 255 amino acids. Mouse EMSP1 shares 75% amino acid identity with pig EMSP1 and has three potential N-linked glycosylation sites, two of which are conserved in the pig homologue. Western blot analysis shows that the polyclonal antibodies are specific for EMSP1 and do not cross-react with trypsin. Immunohistochemistry of pig incisors shows discrete staining in the surface enamel at the earliest part of the maturation stage. In mouse molars, in situ hybridization gives a distinct and specific signal in maturation-stage ameloblasts, and in the junctional epithelium following tooth eruption. We conclude that EMSP1 is expressed by pig and mouse ameloblasts during the early maturation stage of amelogenesis.
The Expression of MMP-8 in Human Odontoblasts and Dental Pulp Cells is Down-regulated by TGF-β1
Authors: Palosaari H. , Wahlgren J. , Larmas M. , Rönkä H. , Sorsa T. , Salo T. , Tjäderhane L.
Published: January 2009

Recent findings show that matrix metalloproteinase-8 (MMP-8) is expressed, in addition to neutrophils, by human chondrocytes, cultured fibroblasts, and endothelial cells. We investigated the expression of MMP-8 in other human mesenchyme-derived cells, odontoblasts, and pulp tissue. Odontoblasts and pulp tissue were collected from extracted human teeth for MMP-8 mRNA analysis with reversetranscription/polymerase chain-reaction (RT-PCR) and Southern blot. The expression, localization, and secretion of MMP-8 protein were studied with Western blot, immunohistochemistry, and immunofluorometric assay. The effect of TGF-β1 (10 ng/mL) on the expression, secretion, and concentration of secreted MMP-8 was studied by odontoblast and pulp tissue culture methods (Tjaderhane et al., 1998a). RT-PCR demonstrated MMP-8 mRNA expression in native and cultured odontoblasts and pulp tissue and cultured pulp fibroblasts, with a 522-bp transcript comparable with that of bone marrow cells. The specificity of PCR was confirmed with Southern blot. Western blot with MMP-8-specific antibody detected 65- and 50-kDa proteins in native samples, representing latent and active forms of mesenchymal-type MMP-8, and in the conditioned odontoblast culture media, 50-kDa protein was observed. TGF-β down-regulated the MMP-8 mRNA and concentration of secreted protein in both cultures. Immunohistochemical staining detected MMP-8 in odontoblasts. These findings indicate that mesenchyme-derived cells of the dentin-pulp complex express, synthesize, and activate MMP-8, which may, in concert with odontoblast-derived gelatinases, participate in organization of dentin organic matrix prior to mineralization.
Effects of Organic Acid Anions on Growth, Glycolysis, and Intracellular pH of Oral Streptococci
Authors: Dashper S.G. , Reynolds E.C.
Published: January 2009

Oral streptococci produce large quantities of organic acids as the end-products of carbohydrate fermentation. In an approach to determine if oral streptococci exhibit differential sensitivities to organic acid anions, we determined the effects of formate, lactate, and acetate on intracellular pH maintenance, glycolysis, and growth of Streptococcus mutans and Streptococcus sanguis. Growth was determined as maximum culture optical density in the presence of the organic acid anions at pH 7.1, 6.7, 6.3, and 6.1, and the effects of the anions on glycolytic activity and intracellular pH were determined at pH 7.0 and 5.0. At pH 7.1, the organic acid anions had little effect on growth of either species. At the lower pH values, all of the anions reduced the maximum culture optical density of both species in a pH- and concentration-dependent manner, with S. sanguis being more sensitive to growth inhitition than S. mutans. The organic acid anions had little or no effect on glycolytic activity of either species at pH 7.0. However, all of the organic acid anions tested reduced glycolytic activity at pH 5.0 in a concentration-dependent manner, with S. sanguis being more sensitive than S. mutans. The inhibition of glycolysis could be related to the pKa of the organic acid, with formate and lactate being more inhibitory than acetate. The organic acid anions decreased the intracellular pH of S. mutans and S. sanguis, glycolyzing at an extracellular pH of 5.0, such that the reduction in glycolytic activity caused by the organic acid anions could be directly attributed to the fall in intracellular pH. In conclusion, the production of lactic acid in plaque would not only lower pH, thereby having a disadvantageous effect on less aciduric oral streptococci, such as S. sanguis, but would also increase their sensitivity to the effects of low pH, helping S. mutans to become more dominant.
Adenoviral-mediated Gene Transfer to Mouse Salivary Glands
Authors: Wang S. , Baum B.J. , Yamano S. , Mankani M.H. , Sun D. , Jonsson M. , Davis C. , Graham F.L. , Gauldie J. , Atkinson J.C.
Published: January 2009

Adenoviral vectors effectively transfer genes to rat salivary glands. However, potent immune responses limit their use in vivo. Mice offer more opportunities than rats for the study of these immune processes. We first established conditions for infection of mouse salivary glands, with an adenoviral vector. The effects of time, viral dose, viral diluent buffer volume, and dexamethasone on expression of a transgene, luciferase, were determined by means of the recombinant vector AdCMVluc. Optimal luciferase expression was observed when the vector was suspended in 50 μL of buffer. This volume completely filled the gland parenchyma and slightly distended the capsule. Dexamethasone increased immediate transgene expression and reduced the acute inflammation one day following viral administration, but did not alter subsequent mononuclear inflammation or transgene expression 14 or 28 days later. An adenoviral vector encoding either antiinflammatory cytokine IL-4 or IL-10 was co-administered with AdCMVluc to increase transgene expression at 14 and 28 days. While this strategy did not extend the duration of luciferase expression, co-administration of AdCMVIL-10 with AdCMVluc almost completely eliminated the chronic inflammatory infiltrate in the glands after 28 days. This study demonstrates that adenoviral-mediated gene transfer to mouse submandibular glands is possible by intraductal cannulation and that reduction of either the acute or chronic inflammatory infiltrates was insufficient to increase long-term transgene expression in this tissue.
Evidence of Chemical Bonding at Biomaterial-Hard Tissue Interfaces
Authors: Yoshida Y. , Van Meerbeek B. , Nakayama Y. , Snauwaert J. , Hellemans L. , Lambrechts P. , Vanherle G. , Wakasa K.
Published: January 2009

For many years, glass-polyalkenoate cements have been described as possessing the unique properties of self-adherence to human hard tissues, such as bones or teeth. However, direct experimental evidence to prove the existence of chemical bonding has not been advanced. X-ray Photoelectron Spectroscopy (XPS) was used to analyze the chemical interaction of a synthesized polyalkenoic acid with enamel and synthetic hydroxyapatite. For both enamel and hydroxyapatite, the peak representing the carboxyl groups of the polyalkenoic acid was detected to have significantly shifted to a lower binding energy. De-convolution of this shifted peak disclosed two components with a peak representing unreacted carboxyl groups and a peak suggesting chemical bonding to hydroxyapatite. On average, 67.5% of the carboxyl groups of the polyalkenoic acid were measured to have bonded to hydroxyapatite. XPS of hydroxyapatite also disclosed its surface to be enriched in calcium and decreased in phosphorus, indicating that phosphorus was extracted at a relatively higher rate than calcium. Analysis of these data supports the mechanism in which carboxylic groups replace phosphate ions (PO 43-) of the substrate and make ionic bonds with calcium ions of hydroxyapatite. It is concluded that an ultrathin layer of a polyalkenoic acid can be prepared on a hydroxyapatite-based substrate by careful removal of non-bonded molecules. With this specimen-processing method, XPS not only provided direct evidence of chemical bonding, but also enabled us to quantify the percentages of functional groups of the polyalkenoic acids that bonded to calcium of hydroxyapatite.
Lifetime-limiting Strength Degradation from Contact Fatigue in Dental Ceramics
Authors: Jung Y.-G. , Peterson I.M. , Kim D.K. , Lawn B.R.
Published: January 2009

The hypothesis under examination in this paper is that the lifetimes of dental restorations are limited by the accumulation of contact damage during oral function; and, moreover, that strengths of dental ceramics are significantly lower after multi-cycle loading than after single-cycle loading. Accordingly, indentation damage and associated strength degradation from multi-cycle contacts with spherical indenters in water are evaluated in four dental ceramics: "aesthetic" ceramics-porcelain and micaceous glass-ceramic (MGC), and "structural" ceramics—glass-infiltrated alumina and yttria-stabilized tetragonal zirconia polycrystal (Y-TZP). At large numbers of contact cycles, all materials show an abrupt transition in damage mode, consisting of strongly enhanced damage inside the contact area and attendant initiation of radial cracks outside. This transition in damage mode is not observed in comparative static loading tests, attesting to a strong mechanical component in the fatigue mechanism. Radial cracks, once formed, lead to rapid degradation in strength properties, signaling the end of the useful lifetime of the material. Strength degradation from multi-cycle contacts is examined in the test materials, after indentation at loads from 200 to 3000 N up to 106 cycles. Degradation occurs in the porcelain and MGC after ~ 104 cycles at loads as low as 200 N; comparable degradation in the alumina and Y-TZP requires loads higher than 500 N, well above the clinically significant range.
Human Gingival Crevicular Fluid Contains MRP8 (S100A8) and MRP14 (S100A9), Two Calcium-binding Proteins of the S100 Family
Authors: Kojima T. , Andersen E. , Sanchez J.C. , Wilkins M.R. , Hochstrasser D.F. , Pralong W.F. , Cimasoni G.
Published: January 2009

Human gingival crevicular fluid contains unidentified proteins which might play a role as markers in periodontal diseases. Therefore, low-molecular-weight proteins found in human gingival crevicular fluid (GCF), but absent from serum, were identified in the present study by means of two-dimensional electrophoresis (2-D PAGE) analysis. GCF, serum, and whole saliva were collected from periodontitis and healthy subjects, as well as from edentulous and newborn subjects. Protein samples were separated by two-dimensional polyacrylamide gel electrophoresis, stained with silver, and compared with reference protein maps in the SWISS-2D PAGE database. In GCF and saliva from periodontitis patients and healthy subjects, four dominant low-molecular-mass (from 8 to 14 kDa) acidic spots were observed. They were not found in serum and were less visible in saliva from edentulous and newborn subjects. From N-terminal amino acid sequencing, the two 2-D protein spots of 8 kDa and isoelectric points between 6.5 and 7.0 were both identified as protein MRP8 (S100A8), a member of the S100 family of calcium-binding proteins. Using peptide mass fingerprinting and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), we identified the other two protein spots, with mass of 14 kDa and isoelectric points between 5.5 and 6.0, as protein MRP14 (S100A9), also belonging to the S100 family. The presence of MRP8 and MRP14 in GCF was confirmed by Western blot, with monoclonal antibodies. The two polypeptides, MRP8 and MRP14, identified in GCF represent the major difference between the 2-D PAGE patterns of serum and GCF, and we hypothesize that they may play an important role in the gingival sulcus and could represent possible markers for periodontal diseases.
Age, Dental Infections, and Coronary Heart Disease
Authors: Mattila K.J. , Asikainen S. , Wolf J. , Jousimies-Somer H. , Valtonen V. , Nieminen M.
Published: January 2009

Epidemiological and intervention studies have suggested that infections are risk factors for coronary heart disease (CHD). Dental infections have appeared as cardiovascular risk factors in cross-sectional and in follow-up studies, and the association has been independent of the "classic" coronary risk factors. This case-control study aimed at detailed assessment of the dental pathology found in various CHD categories (including elderly patients). Altogether, 85 patients with proven coronary heart disease and 53 random controls, matched for sex, age, geographic area, and socio-economic status, were compared with regard to dental status, assessed blindly with four separate scores, and to the "classic" coronary risk factors (seven of the controls had CHD, and they were not included in the analyses). The dental indices were higher among CHD patients than in the controls, but, contrary to previous studies, the differences were not significant (between the CHD patients and their matched controls or among the different CHD categories). This result could not be explained by potential confounding factors. The participants in the present study were older and had more often undergone recent dental treatment in comparison with subjects in our earlier studies. Age correlated with the severity of dental infections only in the random controls but not in the coronary patients who, although young, already had high dental scores. We believe that the higher age of the participants in the present study is the most likely reason for the results. Other possible explanations include an age-related selection bias among older CHD patients, and the fact that those participating in studies like this may have better general health and thus also less severe dental infections. Thus, the role of dental infections as a coronary risk factor varies according to the characteristics of the population studied.
Relationship among Mutans Streptococci, "Low-pH" Bacteria, and lodophilic Polysaccharide-producing Bacteria in Dental Plaque and Early Enamel Caries in Humans
Authors: Van Ruyven F.O.J. , Lingström P. , Van Houte J. , Kent R.
Published: January 2009

Multiple interactions occur among major determinants of dental caries. We have studied the bacterial flora and pH-lowering capacity of the same dental plaques in relation to caries. The findings on the plaque flora are reported here. The buccal surfaces of upper teeth in each subject were selected for study. A low-caries group had no "white spot" caries (ws) in the selected dentition area; a higher-caries group averaged 4.1 ws in this area. The latter group was divided into subjects with 2, 3, or 4 ws and subjects with 5, 6, or 7 ws. Enumerated organisms in plaque samples (sound and ws sites) from all subjects were: (1) mutans streptococci (MS) on mitis-salivarius-bacitracin and mitis-salivarius agar; (2) non-mutans streptococci (non-MS) on mitis-salivarius agar; (3) organisms that were categorized according to their minimum pH in sugar broth, i.e., the predominant undifferentiated total flora on blood agar or the predominant non-MS flora on mitis-salivarius agar; and (4) iodophilic polysaccharide-storing organisms on trypticaseyeast extract-salts agar. Plaques covering ws lesions contained generally only low proportions (< 0.1%) of MS. The plaque proportions of all the above 4 bacterial groups were increased in the higher-caries group but were similar for s and ws sites in this group. Over half of the total plaque flora in subjects with 5, 6, or 7 ws consisted of "low-pH"-type organisms (minimum pH < 4.4). Many of these were neither MS nor "low-pH" non-MS. The numerical emergence of MS in plaque appeared to be preceded often by other types of "low-pH" bacteria, including the non-MS. Caries development in the absence or presence of MS as well as different bacterial successions in plaque can be explained readily by the dynamic and positive relationship among the factors carbohydrate consumption, plaque flora composition, plaque acidogenic potential, and caries activity.
Development and Characterization of a Tissue-engineered Human Oral Mucosa Equivalent Produced in a Serum-free Culture System
Authors: Izumi K. , Terashi H. , Marcelo C.L. , Feinberg S.E.
Published: January 2009

A problem maxillofacial surgeons face is a lack of sufficient autogenous oral mucosa for reconstruction of the oral cavity. Split-thickness or oral mucosa grafts require more than one surgical procedure and can result in donor site morbidity. Skin has disadvantages of adnexal structures and a different keratinization pattern than oral mucosa. In this study, we successfully assembled, ex vivo, a human oral mucosa equivalent, consisting of epidermal and dermal components, in a defined, essential-fatty-acid-deficient, serum-free culture medium without a feeder layer, that could be used for intra-oral grafting in humans. Autogenous oral keratinocytes were seeded onto a cadaveric dermis, AlloDerm™. The oral mucosa equivalent was cultured at an air-liquid interface for 2 wks. The resulting equivalent had a well-stratified parakeratinized epithelial layer similar to native oral keratinized mucosa. Expression of differentiation markers, filaggrin and cytokeratin 10/13, suggested a premature keratinized state. The presence of proliferation markers, proliferating cell nuclear antigen (PCNA) and Ki-67, suggested a state of hyperproliferation. Fatty acid composition of the equivalent was similar to that of in vitro cultured oral keratinocytes but differed from the that of in vivo native tissue, showing a lower content of 18:2 and 20:4, and a higher content of 16:1 and 18:1 fatty acids, respectively. The keratinocytes of the equivalent appeared to be in a more active and proliferative state than native keratinized mucosa. The dynamic nature of the cell population on the oral mucosa equivalent may be beneficial for intra-oral grafting procedures and for transfection of the keratinocytes.
The Effects of Adhesive Thickness on Polymerization Contraction Stress of Composite
Authors: Choi K.K. , Condon J.R. , Ferracane J.L.
Published: January 2009

A layer of an unfilled adhesive resin placed between the tooth and composite restoration has been shown to absorb some of the stress generated in the composite during polymerization and to reduce interfacial leakage. The objectives of this study were to measure the change in polymerization contraction stress of bonded composite as the thickness of the resin adhesive was systematically varied, and to correlate the effects of the adhesive thickness and reduced stress on marginal leakage in class V cavities. The maximum contraction force of composite (Herculite XRV) was measured in a tensilometer as the thickness of the adhesive bonding agent (Scotchbond MP) was varied from 20 to 300 μm. Composite was placed in Class V cavities prepared on the labial surfaces of bovine teeth to which different thicknesses of adhesive had been applied by layering, and a marginal leakage test was performed by means of staining with silver nitrate. Contraction stress decreased significantly as the adhesive thickness was increased. This result was supported by a theoretical examination of the data. In class V cavities, additional adhesive layering in the marginal area reduced the overall degree of microleakage. The contraction stress generated during the placement of a composite restoration contributes significantly to early marginal leakage, and this stress was significantly absorbed and relieved by the application of an increasing thickness of low-stiffness adhesive.
Mass Spectrometry of Native Rat Amelogenins: Primary Transcripts, Secretory Isoforms, and C-terminal Degradation
Authors: Chen W.-Y. , Bell A.W. , Simmer J.P. , Smith C.E.
Published: January 2009

Cloning technologies have established unambiguously that amelogenins always seem larger in molecular weight ( Mr) by gel electrophoresis (SDS-PAGE) than by mass spectrometry (MS). This has caused many problems relating cloned versions of amelogenin to proteins actually secreted by ameloblasts in vivo. In this study, discrete protein fractions at 31-20 kDa ( Mr SDS) were prepared from freeze-dried rat incisor enamel by techniques optimized for preserving protein integrity. N-terminal sequence and amino acid compositional analyses indicated that the major protein forming these fractions was amelogenin. As expected, the molecular weights estimated by matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) MS were significantly less than their apparent molecular weights estimated by SDS-PAGE. Plots of MrSDS vs. M rMS for all fractions showed high linear correlation (r = 0.992). Analysis of MS data further indicated that the major protein in the 27-kDa fraction corresponded to the R180 secretory isoform of rat amelogenin, whereas some minor proteins in the 23-kDa fraction likely corresponded to a R156 secretory isoform. This was in contrast to major proteins forming the 25-, 24-, and 23-kDa fractions ( MrSDS), which seemed to represent proteolytic fragments of R 180 progressively altered at the P169-A170, P164-L165, and F151-S152 C-terminal cleavage sites, respectively. Proteins in the 20-kDa fraction (MrSDS) most closely matched by ESI-MS fragments of the R156 secretory isoform that were C-terminally-modified at the equivalent P164-L165 site.
AgNOR are Sensitive Markers of Radiation Lesions in Squamous Epithelia
Authors: lelmini M.V. , Heber E. , Schwint A.E. , Cabrini R.L. , Itoiz M.E.
Published: January 2009

The possibility of detection of incipient cellular alterations is central to early diagnosis and to clinician's capacity to discriminate between samples that appear similar on routine preparations. We examined the value of silver-stained nucleolar organizer regions (AgNOR) in detecting radio-induced alterations in a model of squamous epithelium biologically similar to oral mucosa. Morphometry of AgNOR has been proven to be of value in the detection of incipient cellular alterations. This method allows for the quantitative evaluation of lesions induced by high doses of radiation long before they become apparent in routine preparations. We herein examine the capacity of AgNOR to reveal the response to low doses of radiation, closer to the therapeutic or accidental dose to which the epithelium of oral mucosa may be exposed.
Clinical and Genetic Analysis of a Large North European Caucasian Family Affected by Early-onset Periodontitis
Authors: Hodge P.J. , Teague P.W. , Wright A.F. , Kinane D.F.
Published: January 2009

Genetic studies of early-onset periodontitis (EOP) are hampered by several factors. These include delayed onset of the trait, an upper age limit of expression of the disease, and lack of phenotypic information for edentulous family members. Segregation analyses of families with EOP support a major locus hypothesis but fail to define clearly the criteria used for diagnosis of the relatives. Confirmation of a proposed mode of inheritance and the identification of risk genes is awaited by means of family linkage studies. It is suggested that a system can be developed for the current and retrospective diagnosis of relatives of EOP probands. In addition, it is hypothesized that the large family presented here is suitable for a linkage study. Relatives of the proband who were unavailable for a full periodontal examination, were edentulous, or were deceased, were diagnosed by means of documented clinical evidence of periodontal disease or from reported case histories. Segregation analysis was performed. Analysis of the power of the pedigree to detect linkage was carried out by means of the SIMLINK program. Three different categories were defined according to the reliability of diagnosis of EOP. Segregation analysis indicated either autosomal-dominant or X-linked-dominant inheritance in this family. The simulations showed lod scores above 3.0 for all locations of the disease gene, and for each category of diagnosis. In conclusion, a method has been developed which can be used for the diagnosis of relatives of EOP probands when ideal clinical data are unavailable. The simulations suggest that this family is suitable for a genetic linkage study with the aim of identifying the location of one or more susceptibility genes.
Diagnostic Value of a Chelating Agent in Patients with Symptoms Allegedly Caused by Amalgam Fillings
Authors: Vamnes J.S. , Eide R. , Isrenn R. , Höl P.J. , Gjerdet N.R.
Published: January 2009

The chelating agent 2,3 dimercaptopropane-1-sulfonate (DMPS) has been used in a mercury mobilization test for diagnoses in illnesses allegedly associated with the presence of amalgam restorations. DMPS is an accepted antidote to heavy metal poisoning. The aim of the present study was to evaluate the diagnostic value of DMPS in patients with symptoms self-related to their amalgam fillings. The subjects consisted of four groups: 19 healthy controls without amalgam experience; 21 healthy controls with amalgam fillings; 20 patients who claimed symptoms of "mercury poisoning" from dental amalgam; and 20 patients who had amalgam fillings removed because of such symptoms. DMPS (2 mg/kg body weight) was injected intravenously, and urine was collected prior to the injection, 30 and 120 min after the injection, and throughout the next 22 hrs. The samples were analyzed for total mercury by cold vapor atomic absorption spectrophotometry. The total median amounts of mercury excreted over 24 hrs for those with complaints allegedly associated with amalgam and for the healthy controls with amalgam fillings were similar. Persons with amalgam fillings excreted about three times more mercury than those without. The controls, who had never had amalgam fillings, and the subjects who had had their fillings removed excreted median amounts of 8.5 μg and 7.2 μg mercury, respectively. The present DMPS challenge test did not differentiate between patients with or those without complaints self-related to their amalgam fillings but did confirm the higher mercury values in patients with dental amalgam.
Pellicle Precursor Protein Crosslinking: Characterization of an Adduct between Acidic Proline-rich Protein (PRP-1) and Statherin Generated by Transglutaminase
Authors: Yao Y. , Lamkin M.S. , Oppenheim FG
Published: January 2009

Recent work with oral transglutaminase indicated that this enzyme, derived from oral epithelial cells, crosslinked pellicle precursor proteins which may be important in the formation of the acquired enamel pellicle. The purpose of this study was to investigate whether purified acidic PRP-1 can form crosslinks with statherin, and whether such a crosslink is derived from a transglutaminase-catalyzed reaction between glutaminyl and lysyl side-chains, leading to a covalent bond formation. Enzymatic reaction products were analyzed by SDS-PAGE and reverse-phase HPLC. The SDS electrophoretogram revealed a protein band with an apparent molecular weight of 32 kDa, which is consistent with the combined apparent molecular weight of acidic PRP-1 (24 kDa) and statherin (8 kDa). A reaction product isolated by HPLC was characterized by amino acid analysis, which showed a stoichiometry consistent with being an adduct composed of one molecule of acidic PRP-1 and one molecule of statherin. In negative control experiments, it could be shown that this adduct was not detected when the lysines of both substrates were modified by reductive methylation prior to the enzymatic reaction. In addition, amino acid analysis and mass spectrometry confirmed the presence of a γ-glutamyl-∈-lysine dipeptide after enzymatic hydrolysis and the absence of this dipeptide after acid hydrolysis. Analysis of the data obtained indicates that oral transglutaminase is capable of crosslinking acidic PRP-1 and statherin in vitro. In addition, this finding exemplifies the potential of post-secretory processing of salivary proteins, which may represent an additional mechanism to generate new protein species.
EMG Power Spectrum and Motor Unit Characteristics in the Masseter Muscle of the Rabbit
Authors: Turkawski S.J.J. , Van Eijden T.M.G.J.
Published: January 2009

Masticatory muscles contain a large variety of motor units with different physiological and morphological properties. In this study, we tested the hypothesis that a relationship exists between the mechanical and myo-electric properties of single motor units in the masseter muscle of the rabbit. It was expected that faster-contracting motor units, which usually have a relatively large number of fibers with large diameters, should have faster action potentials with larger amplitudes than slower motor units. Single motor units were stimulated. A two-dimensional force transducer registered mechanical parameters of the units. EMG electrodes were used to determine amplitude and frequency parameters of the action potentials of the same units. The results showed that faster-contracting motor units indeed produced action potentials with higher conduction velocities. However, faster motor units had no significant larger amplitude of the action potential. Small but significant positive correlations were found between the tetanic peak force and the amplitude of the action potentials. Little difference was found among the various frequency and amplitude parameters, respectively, making them equally suitable to describe the action potential. Surprisingly, a negative correlation between the amplitude and frequency parameters of the action potential was found, which may result from variability in arrival times of action potentials at the electrode site. Regional differences in the frequency parameters were found between the anterior and posterior parts of the superficial masseter.
The Association of Basic Proline-rich Peptides from Human Parotid Gland Secretions with Caries Experience
Authors: Ayad M. , Van Wuyckhuyse B.C. , Minaguchi K. , Raubertas R.F. , Bedi G.S. , Billings R.J. , Bowen W.H. , Tabak L.A.
Published: January 2009

To address whether there are associations between the peptide composition of human parotid saliva and dental decay (caries) experience, we have characterized the peptides from parotid ductal saliva collected from nine adults who have remained free from dental caries (mean age = 59.2; Decayed Missing Filled Surfaces index [DMFS] = 0) and nine individuals who have experienced caries (mean age = 51.2; mean DMFS = 38.4). Ethanol-soluble peptides were size-fractionated on columns of Bio-Gel P-2 ; the salivary peptides derived from caries-susceptible subjects appeared larger than those found in the saliva of caries-free subjects. Peptides were then resolved into 19 species by cation exchange HPLC. Sequence analysis identified 18 peptides that appear to be proteolytic cleavage products of the basic proline-rich proteins IB-4, IB-5, IB-7, IB-8b, and P-B. The peptides that were more abundant in saliva obtained from the caries-free group differed from those isolated from the caries-susceptible group. The median peptide concentration of one possible precursor protein, IB-7, was found to be higher in saliva collected from caries-free individuals than in that from caries-susceptible individuals. Although differences were found in the phenotypes of proline-rich proteins expressed by these groups of caries-free and caries-susceptible subjects, no statistically significant associations were observed among proline-rich phenotypes and the level of any peptide. Collectively, our results indicate that proteolytic processing of parotid salivary proteins differs among individuals who have remained caries-free and those who have experienced dental decay.
Effect of in vitro Acidification on Plaque Fluid Composition With and Without a NaF or a Controlled-release Fluoride Rinse
Authors: Vogel G.L. , Zhang Z. , Chow L.C. , Carey C.M. , Schumacher G.E. , Banting D.W.
Published: January 2009

Plaque fluid ion concentration changes, especially fluoride, in response to the pH decrease associated with a cariogenic episode are important components of the caries process. A "controlled-release" (CR) fluoride rinse, based on the controlled release of fluoride in the presence of calcium, has been shown to form large fluoride reservoirs in resting plaque. In this study, the in vitro acid-induced release of fluoride, and other ions, was examined in 48-hour-fasted plaque fluid from subjects (n = 11) who received no rinse, or who used a 228-ppm CR or NaF fluoride rinse 1 hr before being sampled. After collection, the plaque was centrifuged to yield plaque fluid, acidified (0.1 μL of 0.5 mol/L HCl per milligram plaque), and then re-centrifuged before a second sample was obtained. Although previous studies indicated a higher plaque fluid fluoride after the new rinse relative to NaF, no statistically significant difference was observed here. Average fluoride release after acidification (average pH, 5.2) was statistically greater following the use of the CR rinse (153 μmol/L) compared with the NaF rinse (17 μmol/L). No fluoride release was seen in the no-rinse samples. The pH, free calcium, phosphate, acetate, propionate, and buffer capacity were not affected by the different amounts of fluoride deposited in the plaque. However, following acid addition, an increase in free calcium and phosphate was observed, which was also independent of the rinse. The large release of fluoride following acidification suggests that the new rinse may provide an improved cariostatic effect.
Remineralization Effects of a Two-solution Fluoride Mouthrinse: An in situ Study
Authors: Chow L.C. , Takagi S. , Carey C.M. , Sieck B.A.
Published: January 2009

Results from previous studies show that a two-solution fluoride (F) rinse is significantly more effective than a NaF rinse of the same F content of 250 μg/g (ppm) in remineralizing enamel and root lesions in an in vitro cyclic de- and remineralization model. In the present study, the two-solution rinse and two NaF rinses with F contents of 250 ppm and 1000 ppm were evaluated in an intra-oral remineralization model. Caries-like lesions were formed in the enamel of extracted human molars with the use of a pH 4 demineralizing solution. Thin sections of the enamel (~ 120 μm) containing lesions were prepared, and the mineral contents of the lesions were assessed by quantitative microradiography. With the cut surfaces protected by nail varnish, 3 enamel specimens were mounted with wax in the lingual areas of a removable mandibular appliance. The study used a randomized, crossover design with seven subjects. In each of the 3 legs of the study, subjects wore the appliances continuously except when eating, drinking, and brushing their teeth. Twice daily (after breakfast and before bedtime), the subjects received a one-minute rinse with 20 mL of (1) 250-ppm-F NaF rinse, (2) 1000-ppm-F NaF rinse, or (3) 228-ppm-F two-solution F rinse. At the end of the 14-day experimental period, the sections were retrieved, and the mineral contents of the lesions were again assessed quantitatively. The results show that both the 1000-ppm-F NaF and 228-ppm-F two-solution rinses produced a greater (p < 0.05) remineralization than did the 250-ppm-F NaF rinse. The remineralization produced by the two-solution rinse was not statistically different (p > 0.05) from that produced by the NaF rinse with 4x the F content (1000 ppm F).
Water-insoluble Glucan Synthesis by Mutans Streptococcal Strains Correlates with Caries Incidence in 12- to 30-month-old Children
Authors: Mattos-Graner R.O. , Smith D.J. , King W.F. , Mayer M.P.A.
Published: January 2009

Early mutans streptococci (MS) infection has been associated with higher caries activity in childhood. Since colonization with MS does not always lead to caries activity, additional factors may be involved in MS cariogenicity. For example, MS may differ in virulence traits such as the potential to synthesize glucan polymers from sucrose. In the present study, we tested the hypothesis that caries activity can be associated with variations in virulence factor expression of MS-infecting strains. At baseline, levels of MS obtained by the tongue-blade sampling method, and the presence of visible plaque on upper incisors, were measured in 101 12- to 30-month-old children. Dental caries lesions were diagnosed at baseline and after one year. Caries incidence data were then used to select ten caries-free and nine caries-active children from whom a total of 20 MS fresh isolates was studied. Water-insoluble glucan (WIG) synthesis, final pH, and sucrose-dependent adherence on glass surfaces were measured in these MS isolates. Concentrated culture supernatants were separated in duplicate SDS-PAGE gels, which were then either stained for protein or incubated with 5% sucrose. The intensities of the WIG bands developed in the 5% sucrose PAGE gels and the corresponding protein-stained GTF bands were measured by scanning densitomctry. High MS levels (≥ 100 CFU) were associated with high caries incidence (p < 0.01). The presence of visible plaque did not correlate with caries incidence. The intensities of WIG bands were positively correlated with caries incidence (p < 0.05) and with the ability of MS to adhere to glass surfaces (p < 0.05). Analysis of our data suggests that the ability to synthesize WIG is an important virulence factor in initial caries development by increasing MS adherence and accumulation in the plaque of young children.
Crack Initiation Modes in Bilayered Alumina/Porcelain Disks as a Function of Core/Veneer Thickness Ratio and Supporting Substrate Stiffness
Authors: Wakabayashi N. , Anusavice K.J.
Published: January 2009

We hypothesize that the fracture resistance of alumina core/porcelain veneer disks increases and that crack initiation shifts from veneer to core as the core/veneer thickness ratio (tc/tv) increases from 0.5/1.0 to 1.3/0.2, or as the elastic modulus of the supporting substrate (Es) to which it is resin-bonded increases from 5.1 to 226 GPa. When supported by a low-modulus substrate, disks with low tc/tv ratios exhibited cracks in the veneer and within the core, while those with high tc/tv ratios demonstrated core cracks, but not veneer cracks. None of the disks supported by Ni-Cr alloy (E = 226 GPa) exhibited core cracks. These results support the hypothesis that the crack initiation site shifts as the tc/t v ratio increases, but the increase in Es did not affect the crack initiation site. This study suggests that the tc/tv ratio is the dominant factor that controls the failure initiation site in bilayered ceramic disks.
PTCH Gene Mutations in Odontogenic Keratocysts
Authors: Barreto D.C. , Gomez R.S. , Bale A.E. , Boson W.L. , De Marco L.
Published: January 2009

An odontogenic keratocyst (OKC) is a benign cystic lesion of the jaws that occurs sporadically or in association with nevoid basal cell carcinoma syndrome (NBCCS). Recently, the gene for NBCCS was cloned and shown to be the human homologue of the Drosophila segment polarity gene Patched ( PTCH), a tumor suppressor gene. The PTCH gene encodes a transmembrane protein that acts in opposition to the Hedgehog signaling protein, controlling cell fates, patterning, and growth in numerous tissues, including tooth. We investigated three cases of sporadic odontogenic keratocysts and three other cases associated with NBCCS, looking for mutations of the PTCH gene. Non-radioactive single-strand conformational polymorphism and direct sequencing of PCR products revealed a deletion of 5 base pairs (bp) in exon 3 (518delAAGCG) in one sporadic cyst as well as mutations in two cysts associated with NBCCS, a nonsense (C2760A) and a missense (G3499A) alteration. This report is the first to describe a somatic mutation of PTCH in sporadic odontogenic keratocysts as well as two novel mutations in cysts associated with NBCCS, indicating a similar pathogenesis in a subset of sporadic keratocysts.
Influence of Environmental pH on the Reactivity of Candida albicans with Salivary IgA
Authors: Bikandi J. , Moragues M.D. , Quindós G. , Polonelli L. , Pontón J.
Published: January 2009

Salivary secretory IgA reacts with a group of heat-shock mannoproteins preferentially expressed on Candida albicans yeast cells and germ tubes grown at 37°C. Since other environmental factors can also modulate the expression of those antigens, we have investigated the influence of the pH of the culture medium on the expression of the antigens reacting with human salivary IgA by C. albicans. By indirect immunofluorescence, yeast cells grown in Sabouraud glucose broth at 37°C showed a statistically significant increase in reactivity with salivary IgA (p < 0.0001) when compared with cells grown at 25°C at the 4 pH values studied (3.3, 5.9, 7.5, and 9.5), the highest reactivity and the major heat-shock effect being observed at pH 5.9. The decrease in reactivity with salivary IgA observed in C. albicans cells grown at pH values of 3.3 and 9.5 was confirmed by Western blotting. Salivary IgA reacted with polydispersed materials from the cell walls of molecular masses > 55 kDa, which were more expressed at neutral pH than at acidic or alkaline pH values. A similar reactivity was observed when the antigenic extracts were stained with an antiserum directed against oligosaccharides present in antigen 6 of C. albicans serotype A. The differences in reactivity presented by salivary IgA may be related to a decrease in the expression of polysaccharides present on the surfaces of the yeast cells of C. albicans grown at acidic or alkaline pH values. The low reactivity of salivary IgA with C. albicans cells grown at acidic pH values may help to explain the association between acidic saliva and the carriage of Candida in the oral cavity, as well as with oral candidiasis.
Unique Enamel Phenotype Associated with Amelogenin Gene (AMELX) Codon 41 Point Mutation
Authors: Ravassipour D.B. , Hart P.S. , Hart T.C. , Ritter A.V. , Yamauchi M. , Gibson C. , Wright J.T.
Published: January 2009

Different mutations in the amelogenin gene (AMELX) result in the markedly different enamel phenotypes that are collectively known as amelogenesis imperfecta (AI). We hypothesize that unique phenotypes result from specific genetic mutations. The purpose of this study was to characterize the enamel compositional and structural features associated with a specific AMELX mutation in three families with X-linked AI. We performed mutational analysis by amplifying AMELX exons and sequencing the products. Permanent and primary affected (N = 6) and normal (N = 3) teeth were collected and examined by light, scanning, and transmission electron microscopy. Enamel proteins were evaluated by immunolocalization of amelogenin and amino acid analysis. AI-affected individuals all shared a common AMELX point mutation (C to A change at codon 41). The dental phenotypic findings were remarkably consistent in all affected individuals. The AI enamel was opaque, with numerous prism defects or holes encompassing the entire prism width. Affected crystallites appeared more radiolucent and morphologically less uniform, compared with that of normal enamel. Immunogold labeling with anti-amelogenin antibodies localized amelogenin to the crystallites but not to the inter-crystalline spaces. No immunogold labeling was seen in normal enamel. There was an increased and amelogenin-like protein content in AI enamel (0.95%) compared with normal enamel (0.13%). We conclude that this codon 41 C to A missense point mutation, in a highly conserved region of the AMELX gene, results in a remarkably consistent phenotype.
Proviral HIV-1 DNA in Gingival Crevicular Fluid of HIV-1-infected Patients in Various Stages of HIV Disease
Authors: Matičić M. , Poljak M. , Kramar B. , Tomažić J. , Vidmar L. , Zakotnik B. , Skalerič U.
Published: January 2009

The oral cavity is rarely reported to be a site of human immunodeficiency virus (HIV) transmission, despite detectable virus in saliva and relatively frequent prevalence of periodontal disease in HIV-infected persons yielding increased excretion of mononuclear-cell-enriched gingival fluid. To search for possible sources of HIV in saliva, and using the polymerase chain-reaction technique, we sought the presence and shedding patterns of proviral HIV-1 DNA in gingival crevicular fluid in a group of patients previously determined as HIV-1-seropositive. Periodontal status at the collection sites was monitored by several clinical parameters, including Plaque Index, Gingival Index, probing depth, and clinical attachment loss. Gingival crevicular fluid samples were collected by means of paper points. Proviral HIV-1 DNA was detected in the gingival fluid of 17 out of 35 HIV-1-infected patients. Its detection correlated significantly with higher plasma HIV-1 RNA viral load (p = 0.03) and not with peripheral blood CD4+ cell count, the presence of blood in gingival fluid, or oral lesions. There was a significant correlation between clinical attachment loss at the sites of fluid collection and plasma HIV-1 RNA viral load (p = 0.002), and borderline correlation between the latter and probing depth (p = 0.54) in the group of patients harboring proviral HIV-1 DNA in gingival crevicular fluid. The results of our study suggest that mononuclear cells present in gingival crevicular fluid and harboring proviral HIV-1 DNA could represent a potential source of HIV-1 in the presence or absence of local bleeding, especially in persons with advanced HIV infection and increased loss of clinical attachment.