The most important progress in diagnostic sciences is the increased sensitivity and specificity in diagnostic procedures due to the development of micromethodologies and increasing availability of immunological and molecular biological reagents. The technological advances led to consider the diagnostic use of saliva for an array of analytes and DNA source. The purpose of the present study was to compare DNA from saliva with those from blood and buccal swab, to evaluate diagnostic and forensic application of saliva, to investigate the changes of genomic DNA in saliva according to the storage temperature and period of saliva samples, and to evaluate the integrity of the DNA from saliva stored under various storage conditions by PCR analysis. Peripheral venous blood, unstimulated whole saliva, stimulated whole saliva, and buccal swab were obtained from healthy 10 subjects (mean age: $29.9{\pm}9.8$ years) and genomic DNA was extracted using commercial kit. For the study of effects of various storage conditions on genomic DNA from saliva, stimulated whole saliva were obtained from healthy 20 subjects (mean age: $32.3{\pm}6.6$ years). After making aliquots from fresh saliva, they were stored at room temperature, $4^{\circ}C$, $-20^{\circ}C$, and $-70^{\circ}C$. Saliva samples after lyophilization and dry-out procedure were stored at room temperature. After 1, 3, and 5 months, the same experiment was performed to investigate the changes in genomic DNA in saliva samples. In case of saliva aliquots stored at room temperature and dry-out samples, the results in 2 weeks were also included. Integrity of DNA from saliva stored under various storage conditions was also evaluated by PCR amplification analysis of $\beta$-globin gene fragments (989-bp). The results were as follows: 1. Concentration of genomic DNA extracted from saliva was lower than that from blood (p<0.05), but there were no significant differences among various types of saliva samples. Purities of genomic DNA extracted from stimulated whole saliva and lyophilized one were significantly higher than that from blood (p<0.05). Purity of genomic DNA extracted from buccal swab was lower than those from various types of saliva samples (p<0.05). 2. Concentration of genomic DNA from saliva stored at room temperature showed gradual reduction after 1 month, and decreased significantly in 3 and 5 months (p<0.05, p<0.01, respectively). Purities of DNA from saliva stored for 3 and 5 months showed significant differences with those of fresh saliva and stored saliva for 1 month (p<0.05). 3. In the case of saliva stored at $4^{\circ}C$ and $-20^{\circ}C$, there were no significant changes of concentration of genomic DNA in 3 months. Concentration of DNA decreased significantly in 5 months (p<0.05). 4. There were no significant differences of concentration of genomic DNA from saliva stored at $-70^{\circ}C$ and from lyophilized one according to storage period. Concentration of DNA showed decreasing tendency in 5 months. 5. Concentration of genomic DNA immediately extracted from saliva dried on Petri dish were 60% compared with that of fresh saliva. Concentration of DNA from saliva stored at room temperature after dry-out showed rapid reduction within 2 weeks (p<0.05). 6. Amplification of $\beta$-globin gene using PCR was successful in all lyophilized saliva stored for 5 months. At the time of 1 month, $\beta$-globin gene was successfully amplified in all saliva samples stored at $-20^{\circ}C$ and $-70^{\circ}C$, and in some saliva samples stored at $4^{\circ}C$. $\beta$-globin gene was failed to amplify in saliva stored at room temperature and dry-out saliva.
Bioequivalence test of $Asthcontin^{\circledR}$ tablet, a commercial slow-release theophylline (TP) dosage form, was performed using $Slo-bid^{\circledR}$ capsule as the reference. Since it has been confirmed that the saliva concentration of TP is closely correlated with the plasma concentration in man, the area under the saliva concentration-time curve was used as a bioavailability parameter. The statistical analysis showed that the two dosage forms are equivalent in bioavailability estimating from the saliva concentration. The results supported that the use of soliva as a test sample provides simple and easy techniques for bioequivalence tests of TP-containing dosage forms.
Parotid and whole saliva were collected from 27 healthy adults, from 25 years of age to 30, and from 27 patients with oral ulcer, from 23 years of age to 61. The amount of each Salivary immunoglobulin A was measured by single radial immunodiffusion (SRID) technique. Results were as follows : 1. There was no significant difference between the normal group and the disease group in the concentration of immunoglobulin A in whole saliva. 2. The concentration of immunoglobulin A in parotid saliva of the normal group was higher than the disease group and the difference was statistically significant between the two groups. (p<0.01) 3. The concentration of immunoglobulin A of the parotid saliva in both groups was higher than that of the whole saliva.
Objectives: The purpose of the study is to investigate the effect of chewing gum containing Prunus mume extract(PME) on the change of saliva ingredients. On the basis of the biological background of molecules and diagnostic indices in the use of saliva, the mastication effect of chewing gum containing PME was demonstrated in terms of secretory IgA concentration and total protein concentration in stimulated saliva. Methods: This study is an experimental research on the use of a research design before and after applying a randomized control group. Participants were distributed randomly to the experimental group and the control group, respectively. The experiment group was instructed to masticate the chewing gum containing PME for 10 minutes for one month after each meal within 30 minutes. Salivary secretion was collected by the participants between 8 and 10 a.m in the morning in the research office. For the measurement of secretory IgA and total protein concentrations in the saliva, indirect enzyme-linked immunosorbent assay(ELISA) was used. Results: The salivation stimulation rate was significantly increased after four weeks of masticating chewing gum containing PME after each meal(p<0.001). Mastication of chewing gum containing PME for four weeks decreased the concentration of secretory IgA much more significantly than that after mastication for one week(p=0.003). The concentration of total protein in the saliva was decreased after four weeks in the experimental and control groups. Conclusions: Mastication of chewing gum containing PME stimulated salivary secretion and led to oral disease prevention in patients with xerostomia. Furthermore, it seems to be urgent to seek measures that can be utilized in intervention for patients with xerostomia.
Objectives: The purpose of the study was to investigate the effect of calcium concentration in saliva on dental caries activity after consuming calcium. Methods: A total of 59 adult women aged 20 to 40 years were surveyed for calcium intake. The daily average calcium intake was analyzed through dietary records of the subjects. The subjects were divided into two groups based on daily average calcium intake. Salivary pH and concentrations of minerals in the saliva were obtained from A group and B group. Calcium ($Ca^{2+}$) and magnesium ($Mg^{2+}$) concentrations in saliva were measured by HPLC-Ion chromatography using 15 mM sulfuric acid. The dental caries activity test was quantified by salivary buffer capacity test and plaque pH test. Results: The mean $Ca^{2+}$ concentrations of A group was $12.75{\mu}g/m$, the mean $Ca^{2+}$ concentrations in the B group was $16.30{\mu}g/mL$ (p<0.05) and respectively, $Mg^{2+}$ concentrations were found to be $0.48{\mu}g/mL$ and $0.51{\mu}g/mL$. Calcium intake and calcium concentration in saliva showed a significant correlation (r=0.380). Conclusions: The mean $Ca^{2+}$ concentrations in saliva was higher in the high calcium intake group. Therefore, calcium intake in saliva was correlated with dental caries.
Plasma and saliva concentrations of acetaminophen (AAP) were determined at various time points by HPLC after oral administration of AAP tablets (AAP 500 mg) to four healthy male Korean subjects. Saliva concentrations (S) of AAP were significantly correlated with plasma AAP concentrations (P). The S/P ratio of AAP was calculated to be 1.05 (r =0.944, $p<10^{-6}$) for all the data points from the subjects. It showed a little intersubject variation and ranged from 0.89 to 1.46 in each subject. Bioavailability parameters such as AUC, $C_{max}$ and $T_{max}$ which are usually obtained from the plasma concentration data will be predictable approximately by saliva concentration data. Saliva seems to be very convinient and useful samples for the preliminary studies of bioavailability and bioequivalence of AAP preparations, since it can be collected frequently without any painful venipuncture to the subjects, that is inevitable in plasma sampling. Evaluation of the bioavailability of a preparation by saliva samples will reduce the cost, time and safety risk greatly in developing a new drug delivery system for AAP.
ELISA kit for cortisol was developed and then evaluated. Polyclonal antihydrocortisone-3-(o-carboxymethyl)oxime BSA rabbit serum was used to coat the 96-well microplates. The minimum detection limit of the kit was 250pg of cortisol per milliliter. The within-run variation and the day to day variation of the ELISA system were 2.0 and 5.9 at maximum, respectively. The kit was used to determine whether salivary cortisol concentration could replace blood cortisol concentration in dexamathasone suppression test of dogs. Changes of cortisol concentration were measured in serum or saliva after intravenous administration of 0.01mg of dexamethasone per kilogram of body weight. Blood alone, saliva alone or both were collected at 0, 30, 60, 120, 240, and 360 minutes after injection of dexamethasone. The change in blood cortisol concentration was found to be suitable in dexamathasone suppression test of dogs, but the change in salivary cortisol concentration was not. The kit was also used to determine whether salivary cortisol concentration could be a stress index as well as blood cortisol concentration in dogs. Two types of trial were performed to estimate the stress either by blood or salivary cortisol concentration. The first trial was stress experiment by intravenous injection of 0.2IU of PZI-insulin per kilogram body weight. Either blood alone or saliva alone was collected at 0, 30, 60, and 90 minutes after insulin administration. Both blood and salivary cortisol concentration were found to be suitable index in estimating stress from hypoglycemia by injection of insulin. The second trial was stress experiment by electrical irritation. The dogs were irritated with anti-bark device for 10 seconds. Blood was collected before and at 2 and 5 minutes after electrical irritation. Saliva was collected before and at 3 and 6 minutes after electrical irritation. The blood cortisol concentration, but not the salivary cortisol concentration was found to be suitable index in estimating stress from electrical irritation. Cushing syndrome in a dog was also successfully diagnosed with this kit.
It has been assumed that nitrite, one of the precursor of N-nitrosamine, in human saliva must have been formed from salivary nitrate through the action of microorganism in the oral cavity. In this paper, we have tested the concentration of nitrite and nitrate in human saliva and the degrees of nitrate reduction by oral microflora and identified some bacteria which were able to reduce nitrate. The concentration of nitrite and nitrate was 1.7~9.5ppm and 9.0~28.5ppm respectively. The numbers of total bacteria and nitrate reducing bacteria in four korean human saliva sample were 15~63${\times}10^8$ CFU and 1.0~6.0${\times}10^8$ CFU and the main nitrate reducing bacteria were Streptococcus uberis which was presented in large quantities and showed remarkable reductive activity. Lastly, we knowed that N-dimethylnitrosamine was formed by the reaction between dimethylamine and nitrite in the presence of St. uberis in vitro.
Journal of the korean academy of Pediatric Dentistry
/
v.25
no.4
/
pp.691-703
/
1998
Saliva is obviously potential medium to protect the dental caries by not only physical clearing effect, but aggregating action of protein with bacteria. Nevertheless, we still do not understand how the dental caries occur and what brings the individual difference in caries prevalence. In the regards of dental caries prevalence, we hypothesized that the composition of salivary protein might be different from caries susceptible group to caries resistant group. The purposes of this experiment were focused on the molecular analysis of salivary proteins from the subjects who were involved in multiple caries. Electrophoretic analysis was done on the whole saliva collected from the children with and without multiple caries. We found 86.2% of subjects with multiple caries has approximately 120 KDa protein band while 30.4% in the healthy subjects. And the concentration of the total protein on the subjects with multiple caries is significantly higher than that of the healthy group. However, it turned out that the difference of the salivary composition does not affect the bacterial adhesion to hydroxyapatite bead. With regards of enzymes in saliva, the activity of ${\alpha}-amylase$ and lactate dehydrogenase does not have any significant difference between both groups. However, the concentrations of $Na^+\;and\;Cl^-$ in saliva from multiple caries group is higher than that of the control group. Taken all together, it may be concluded that 120 KDa protein in saliva may be associated with the process of dental caries, also the high concentration of protein and $Na^+,\;Cl^-$ in saliva may be linked to dental caries development as a cofactors.
The tumor marker CA 15-3 is one of the most import reliable for metastatic breast cancer monitoring. While it is generally assessed in serum of patients, blood sampling is an invasive method compared to saliva sampling which is simple and could be an alternative to blood according to many studies. The aim of this investigation was to assess the relationship between serum and salivary concentrations of the protein CA 15-3 in patients with breast cancer and healthy asymptomatic volunteers. A case-control study was conducted with 60 women: 29 breast cancer patients from the Maternity Hospital Souissi Rabat (Morocco) and 31 healthy asymptomatic women. The CA 15-3 concentrations in saliva and serum samples were assessed using an enzyme immune assay (EIA kits) and comparison between cases and controls was made by the Mann-Whitney test. The correlation between serum and saliva CA 15-3 concentration was tested using Pearson correlation. The comparison result of CA15-3 concentration in saliva and serum level in cases and controls was not statistically significant (p>0.05). However, the correlation between salivary and serum CA 15-3 concentration was positive and statistically significant (r=0.27, p=0.03). In conclusion, the positive correlation between salivary and serum expression found in our study suggests that saliva could be an alternative to blood sampling to help breast cancer monitoring.
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