Salivary analysis can be used to assess the severity of caries. Of the known salivary proteins, a paucity of information exists concerning the role of proteinase 3 (PR3), a serine protease of the chymotrypsin family, in dental caries. Whole, unstimulated saliva was collected from children with varying degrees of active caries and tested using a Human Protease Array Kit and an enzyme-linked immunosorbent assay. A significantly decreased concentration of salivary PR3 was noted with increasing severity of dental caries (P<0.01); a positive correlation (r=0.87; P<0.01; Pearson's correlation analysis) was also observed between salivary pH and PR3 concentration. In an antibacterial test, a PR3 concentration of 250 ng·mL⁻¹ or higher significantly inhibited Streptococcus mutans UA159 growth after 12 h of incubation (P<0.05). These studies indicate that PR3 is a salivary factor associated with the severity of dental caries, as suggested by the negative relationship between salivary PR3 concentration and the severity of caries as well as the susceptibility of S. mutans to PR3.
Child ; Dental Caries ; enzymology ; Female ; Humans ; Male ; Myeloblastin ; metabolism ; Saliva ; enzymology

OBJECTIVE: To research the value of polymorphism of salivary esterase(Set) in paternity and personal identification. METHODS: Phenotype and genotype of human salivary esterase were detected in 114 liquid saliva samples from the Chinese population by disc electrophoresis and fast blue RR staining assay. RESULTS: The frequency of Set type was F 22.81%, FS 50.88%, S2 6.31%. The estimated gene frequency of SetF was 0.4825 and SetS was 0.5175. The PE was 0.1875 and the DP was 0.6199. CONCLUSION Polymorphism of salivary esterase (Set) was practical in paternity and personal identification.
Esterases/genetics* ; Forensic Anthropology/methods* ; Gene Frequency ; Humans ; Paternity ; Polymorphism, Genetic ; Saliva/enzymology*

Adult ; Biomarkers ; Humans ; Immunoglobulin A, Secretory ; analysis ; Isoenzymes ; analysis ; Male ; Occupational Diseases ; diagnosis ; immunology ; Saliva ; enzymology ; immunology ; Stress, Psychological ; diagnosis ; immunology

BACKGROUND: Saliva is increasingly being used as a specimen for systemic disease as well as for oral health status. Especially, salivary amylase has been studied as an excellent index for psychological stress. Authors evaluated the measurement of salivary amylase activities collected by Salivettes (Sarstedt, Germany). METHODS: Saliva specimens were collected from 13 healthy adults between 10:00 and 11:00 a.m. Participants were asked to gently chew tampons of Salivettes for 1 min. Immediately after collection, all specimens were stored frozen. On the day of testing, they were centrifuged after thawing and diluted with distilled water. Amylase was measured by Dimension RxL Max (Dade Behring Inc., USA). We evaluated precision, linearity, and recovery rate of Salivette. Amylase activities between collection of saliva by Salivette and passive drool were compared, and also the changes of amylase by the storage temperature were evaluated. RESULTS: Intra-run CVs for three levels of amylase were excellent. Between-day CVs and total CVs were good only for mid and high levels. A good linear relationship was found at all diluted levels. Dosing Salivettes with 2 mL, 1.5 mL, and 1 mL yielded sample recovery 85.5+/-2.4%, 82.4+/-1.5%, and 72.2+/-3.1%, respectively and amylase recovery 78.9+/-10.9%, 74.1+/-13.7%, and 37.3+/-26.9%, respectively. Amylase by Salivette and passive drool were correlated well (r=0.757), although they showed a significant difference. Amylase activity was not affected by the storage temperature. CONCLUSIONS: Measurement of salivary amylase using Salivette could be a useful test having good intra-run CVs and linearity. More than 1.5 mL of saliva would be needed to have more than 70% recovery of Salivette.
Adult ; Amylases/*analysis ; Data Interpretation, Statistical ; Female ; Humans ; Male ; Saliva/*enzymology ; Sensitivity and Specificity ; Specimen Handling/*instrumentation/methods ; Stress, Psychological ; Temperature

OBJECTIVETo compare the concentrations of IgA, lactate dehydrogenase, lysozyme and alkaline phosphatase (ALP) in unstimulated (UWS) and stimulated (SWS) whole saliva between children with severe early childhood caries (S-ECC) and children without caries.

METHODSOne hundred and ninety-two children aged from 42 to 54 months were recruited from 11 urban kindergartens in Beijing. The S-ECC group contained 98 children with more than 5 decayed teeth, and the control group contained 94 caries-free children. The age and sex were matched in the two groups. Two milliliter UWS and 2 ml SWS was collected between 9 and 11 a.m. The salivary IgA was measured by immunoturbidimetric technique. The concentrations of lactate dehydrogenase and ALP were measured by continuous monitoring method, while lysozyme was detected by turbidimetric technique. All results for paired observations between unstimulated and stimulated whole saliva were analysed by paired-samples t test.

RESULTSIn both UWS and SWS, the concentrations of IgA, lactate dehydrogenase and lysozyme in S-ECC children were higher than those in caries-free children (P < 0.01), but the concentration of ALP showed no significant difference in SWS between S-ECC children and caries-free children (P > 0.05).

CONCLUSIONSThe presence of early childhood caries may be associated with an increase of IgA, lactate dehydrogenase and lysozyme in unstimulated and stimulated whole saliva.

Alkaline Phosphatase ; analysis ; Child, Preschool ; Dental Caries ; metabolism ; Female ; Humans ; Immunoglobulin A ; analysis ; L-Lactate Dehydrogenase ; analysis ; Male ; Muramidase ; analysis ; Saliva ; enzymology ; immunology

OBJECTIVE: To investigate the correlation between matrix metalloproteinase (MMP)-2/MMP-9 levels and childhood caries, and the saliva levels of MMP-2/MMP-9 among healthy children and those with different degrees of dental caries, both before and after treatment. METHODS: In the study, 368 children aged 3 to 5 years were separated into three groups: severe caries group (112 children), mild caries group (98 children) and caries free group (158 children). The children with severe caries were included in treatment group (83 children) after accepting a comprehensive treatment of caries. MMP-2 and MMP-9 levels were determined by enzyme-linked immunosorbent assay (ELISA) and the data were analyzed by the Statistics Package for Social Science (SPSS 13.0). The differences among severe caries group, mild caries group and caries free group were analyzed by SNK-q (Student Newman Keuls). The severe caries group and treatment group were compared by paired t test. The differences between each group were statistically analyzed. RESULTS: There was no significant difference of the age and gender composition among severe caries group, mild caries group, caries free group and treatment group. The MMP-2 level of severe caries group [(141.3±32.5) μg/L] was higher than those of mild caries group [(107.5±21.3) μg/L] and caries free group [(102.8±18.5) μg/L] (P<0.05). There was no significant difference between mild caries and caries free group (P>0.05). After analysis of 83 children in the treatment group, the level of MMP-2 [(120.1±24.8) μg/L] was lower than before [(144.6±30.3) μg/L] (P<0.05), but was higher than that of caries free group (P<0.05). The MMP-9 levels of severe caries group [(445.8±68.1) μg/L] and mild caries group [(428.6±59.2) μg/L] were higher than that of caries free group [(385.4±60.6) μg/L] (P<0.05), but the difference between severe caries group and mild caries group was not significant (P>0.05). After analysis of 83 children in the treatment group, the alteration of MMP-9 [(432.2±64.7) μg/L] was not significant either (P>0.05). CONCLUSION The saliva levels of MMP-2 and MMP-9 in children with severe caries were higher than those in caries free children, even if the treatment was implemented, which suggests that the MMP-2 and MMP-9 in saliva might be related to the caries in children.
Child, Preschool ; Dental Caries/enzymology* ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Matrix Metalloproteinase 2/analysis* ; Matrix Metalloproteinase 9/analysis* ; Matrix Metalloproteinases ; Saliva/chemistry*

Streptococcus mutans is a common Gram-positive bacterium and plays a significant role in dental caries. Tobacco and/or nicotine have documented effects on S. mutans growth and colonization. Sortase A is used by many Gram-positive bacteria, including S. mutans, to facilitate the insertion of certain cell surface proteins, containing an LPXTGX motif such as antigen I/II. This study examined the effect of nicotine on the function of sortase A to control the physiology and growth of S. mutans using wild-type S. mutans NG8, and its isogenic sortase-defective and -complemented strains. Briefly, the strains were treated with increasing amounts of nicotine in planktonic growth, biofilm metabolism, and sucrose-induced and saliva-induced antigen I/II-dependent biofilm formation assays. The strains exhibited no significant differences with different concentrations of nicotine in planktonic growth assays. However, they had significantly increased (P≤0.05) biofilm metabolic activity (2- to 3-fold increase) as the concentration of nicotine increased. Furthermore, the sortase-defective strain was more sensitive metabolically to nicotine than the wild-type or sortase-complemented strains. All strains had significantly increased sucrose-induced biofilm formation (2- to 3-fold increase) as a result of increasing concentrations of nicotine. However, the sortase-defective strain was not able to make as much sucrose- and saliva-induced biofilm as the wild-type NG8 did with increasing nicotine concentrations. These results indicated that nicotine increased metabolic activity and sucrose-induced biofilm formation. The saliva-induced biofilm formation assay and qPCR data suggested that antigen I/II was upregulated with nicotine but biofilm was not able to be formed as much as wild-type NG8 without functional sortase A.
Amino Acid Motifs ; Aminoacyltransferases ; drug effects ; genetics ; Antigens, Bacterial ; drug effects ; Bacterial Adhesion ; drug effects ; Bacterial Proteins ; drug effects ; genetics ; Biofilms ; drug effects ; Cysteine Endopeptidases ; drug effects ; genetics ; Dose-Response Relationship, Drug ; Humans ; Mutation ; genetics ; Nicotine ; administration & dosage ; pharmacology ; Peptidoglycan ; drug effects ; genetics ; Saliva ; physiology ; Streptococcus mutans ; drug effects ; enzymology ; growth & development ; Sucrose ; pharmacology

Whole gland perfusion technique was applied to rat parotid glands to assess whether amylase affects fluid secretion. Control perfusion without any secretagogue evoked no spontaneous secretion. Carbachol (CCh 1 microM) induced both amylase and fluid secretion with distinctive kinetics. Fluid secretion occurred constantly around 60 microL/g-min, whereas amylase secretion exhibited an initial peak, followed by a rapid decrease to reach a plateau. Isoproterenol (Isop 1 microM) alone did not induce fluid secretion although it evoked amylase secretion as measured in isolated perfused acini. Addition of Isop during CCh stimulation evoked a rapid and large rise in amylase secretion accompanied by small increase in oxygen consumption. Morphological observations carried out by HR SEM and TEM revealed exocytotic profiles following Isop stimulation. CCh stimulation alone seldom showed exocytotic profiles, suggesting a low incidence of amylase secretion during copious fluid secretion. Combined stimulation of CCh and Isop induced both vacuolation and exocytosis along intercellular canaliculi. These findings suggest that control of salivary fluid secretion is independent of the amylase secretion system induced by CCh and/or Isop.
Amylases/metabolism* ; Animal ; Carbachol/pharmacology ; Cholinergic Agonists/pharmacology ; In Vitro ; Isoproterenol/pharmacology ; Male ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Oxygen Consumption/physiology ; Oxygen Consumption/drug effects ; Parotid Gland/ultrastructure ; Parotid Gland/secretion* ; Parotid Gland/enzymology* ; Perfusion ; Rats ; Rats, Wistar ; Saliva/metabolism* ; Sympathomimetics/pharmacology