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.
The purpose of this study is to evaluate the relationship of menstrual cycle and halitosis by measuring the concentrations of Voltile Sulfur Compounds, secretion rate of unstimulated saliva, secretion rate of stimulated saliva and viscosity of saliva during the menstrual cycle. The subjects were 19 female dental students of Yonsei University who had relatively good alignment of the teeth. They hadn't taken antibiotics or oral contraceptive pills during the few months prior to the experiment, and they didn't have any dental caries involving the pulp or periodontal disease. Lady-$Q^{(R)}$(Alpain Korea, Korea), which confirms the ovulation using saliva, was used to find out the menstrual cycle of subjects. Their history was taken and their basal body temperature was measured. On the basis of these data, the amount of Volatile Sulfur Compounds, secretion rate of unstimulated saliva, secretion rate of stimulated saliva, viscosity of saliva were measured during 1 day of the proliferative phase, 3 days of ovulatory phase and 1 day of the luteal phase within the menstrual cycle. The results were as follows : 1. The amount of Volatile Sulfur Compounds, secretion rate of unstimulated saliva, secretion rate of stimulated saliva, and viscosity of saliva showed no statistically significant cyclic change during proliferative phase, ovulatory phase, and luteal phase(p<0.05). 2. Between the secretion rate of unstimulated saliva and secretion rate of stimulated saliva, there was significant correlation during proliferative phase and luteal phase(p<0.05) and there was no significant correlation during ovulatory phase but relatively close result was seen. 3. The amount of Volatile Sulfur Compounds during proliferative phase and luteal phase had statistically significant correlation(p<0.05). 4. Secretion rate of stimulated saliva during proliferative phase and ovulatory phase, proliferative phase and luteal phase, ovulatory phase and luteal phase had significant correlations (p<0.01).
This study is about saliva from a GIGONG's point of view, it's importance, and it's funtion that recently medical world proved. And here is GIGONG training with saliva. The results were summarized as follows. 1. It is detected that saliva have function to prevent symptoms of senility and have effect of an $anti{\sim}cancer$ medically in recent years. So it is said that science proved narrowly that so called a predecessor speak of salive interms of high praise and they already experienced. 2. Similar substance to saliva exist in human body besides nature food, but human body have materials that deal with all situation to occur from every kind of environment and disease. And exactly saliva is an example of it. 3. We cannot turn back sweat, blood, tears and sperm again that is out of human body once, but we can swallow again only saliva. Therefore, we must experience training that swallow saliva to make us healthy by ourselves, and we don't have to commit a foolish act to cough up saliva at random that a predecessor speak of in terms of high praise, and to be second to JUNG(精). 4. Saliva is a kind of antibody or hormone, so I think that we need to make a study of using saliva with a GUIGONG that is an example of way to strengthen immunity of human body.
Purpose: The purpose of this study was to investigate whether various saliva collection methods affect the observed salivary microbiome and whether microbiomes of stimulated and unstimulated saliva and plaque differ in richness and diversity. Methods: Seven sampling methods for unstimulated saliva, stimulated saliva, and plaque samples were applied to six orally and systemically healthy participants. Bacterial 16S ribosomal RNA genes of 10 major oral bacterial species, namely, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Fusobacterium nucleatum, Prevotella intermedia, Prevotella nigrescens, Streptococcus mitis, Streptococcus sobrinus, and Lactobacillus casei, were analyzed by real-time polymerase chain reaction. We comprehensively examined the dependence of the amount of bacterial ribosomal DNA (rDNA), bacterial-community composition, and relative abundance of each species on sample collection methods. Results: There were significant differences in the bacterial rDNA copy number depending on the collection method in three species: F. nucleatum, P. nigrescens, and S. mitis. The species with the highest richness was S. mitis, with the range from 89.31% to 100.00%, followed by F. nucleatum, P. nigrescens, T. denticola, T. forsythia, and P. intermedia, and the sum of the proportions of the remaining five species was less than 1%. The species with the lowest observed richness was P. gingivalis (<0.1%). The Shannon diversity index was the highest in unstimulated saliva collected with a funnel (4.449). The Shannon diversity index was higher in plaque samples (3.623) than in unstimulated (3.171) and stimulated (3.129) saliva and in mouthwash saliva samples (2.061). Conclusions: The oral microbial profile of saliva samples can be affected by sample collection methods, and saliva differs from plaque in the microbiome. An easy and rapid technique for saliva collection is desirable; however, observed microbial-community composition may more accurately reflect the actual microbiome when unstimulated saliva is assayed.
To obtain primary idea on oral bacterium species that are generally present in periodotally healthy Koreans, the oral bacterial flora in the saliva of four periodontally healthy Koreans at different ages (5, 32, 35, 65) was investigated in this study. For this investigation, 16S rRNA gene clone libraries were generated from the saliva of the four healthy Koreans, and 50 clones were randomly selected from each saliva clone library and sequenced. Totally, 37 different kinds of bacterial 16S rRNA gene sequences were identified based on sequence homology search through GenBank database. The 37 kinds of saliva clone sequences were classified to 14 genera and 2 uncultured and 1 unidentified bacteria. Among the 14 identified genera, Streptococcus, Prevotella, and Veillollella were common genera, and Streptococcus was dominant genus that accounted for 7 different species. Among the seven Streptococcus species, S. salivarius appeared as the most common species. More numbers of species belonging to the genera Streptococcus and Prevotella was present in saliva from ages 32 and 35. While saliva from ages 5 and 65 showed more numbers of species belonging to the genera Rothia, including potential pathogenic species. Overall, saliva of a young child and a senior showed higher bacterial diversity than that of young adults.
Since the outbreak of coronavirus disease 2019 (COVID-2019), the infection has spread worldwide due to the highly contagious nature of severe acute syndrome coronavirus (SARS-CoV-2). To manage SARS-CoV-2, the development of diagnostic assays that can quickly and accurately identify the disease in patients is necessary. Currently, nucleic acid-based testing and serology-based testing are two widely used approaches. Of these, nucleic acid-based testing with quantitative reverse transcription-PCR (RT-qPCR) using nasopharyngeal (NP) and/or oropharyngeal (OP) swabs is considered to be the gold standard. Recently, the use of saliva samples has been considered as an alternative method of sample collection. Compared to the NP and OP swab methods, saliva specimens have several advantages. Saliva specimens are easier to collect. Self-collection of saliva specimens can reduce the risk of infection to healthcare providers and reduce sample collection time and cost. Until recently, the sensitivity and accuracy of the data obtained using saliva specimens for SARS-CoV-2 detection was controversial. However, recent clinical research has found that sensitive and reliable data can be obtained from saliva specimens using RT-qPCR, with approximately 81% to 95% correspondence with the data obtained from NP and OP swabs. These data suggest that self-collected saliva is an alternative option for the diagnosis of COVID-19.
Many of the protective functions of saliva can be attributed to the biological, physical, structural, and rheological characteristics of salivary glycoproteins. Therefore, the development of ideal saliva substitutes requires understanding of the rheological as well as biological properties of human saliva. In the present study, we investigated the changes of salivary enzymatic activities by saliva substitutes and compared viscosity of saliva substitutes with human saliva. Five kinds of saliva substitutes such as Moi-Stir, Stoppers4, MouthKote, Saliva Orthana, and SNU were used. Lysozyme activity was determined by the turbidimetric method. Peroxidase activity was determined with an NbsSCN assay. $\alpha$-Amylase activity was determined using a chromogenic substrate, 2-chloro-p-nitrophenol linked with maltotriose. The pH values of saliva substitutes were measured and their viscosity values were measured with a cone-and-plate digital viscometer at six different shear rates. Various types of saliva substitutes affected the activities of salivary enzymes in different ways. Stoppers4 enhanced the enzymatic activities of hen egg-white lysozyme, bovine lactoperoxidase (bLP), and $\alpha$-amylase. Saliva Orthana and SNU inhibited bLP activity and enhanced $\alpha$-amylase activity. MouthKote inhibited $\alpha$-amylase activity. Moi-Stir inhibited the enzymatic activities of bLP and $\alpha$-amylase. The pH values were very different according to the types of saliva substitutes. Stoppers4, MouthKote, and Saliva Orthana showed lower values of viscosity at low shear rates and higher values of viscosity at high shear rates compared with unstimulated and stimulated whole saliva. Moi-Stir and SNU displayed much higher values of viscosity than those of natural whole saliva. Collectively, our results indicate that each saliva substitute has its own biological and rheological characteristics. Each saliva substitute affects the enzymatic activity of salivary enzyme and finally oral health in different ways.
Sunagawa, K.;Ooshiro, T.;Nakamura, N.;Ishii, Y.;Nagamine, I.;Shinjo, A
Asian-Australasian Journal of Animal Sciences
/
v.20
no.1
/
pp.60-69
/
2007
Ruminants eating dry forage secrete large volumes of saliva which results in decreased plasma volume (hypovolemia) and the loss of $NaHCO_3$ from the blood. The present research investigated whether or not hypovolemia and the loss of $NaHCO_3$ from the blood in goats brought about by dry forage feeding actually depresses feed intake and saliva secretion, respectively. The present experiment consisted of three treatments (NI, ASI, MI). In the control treatment (NI), a solution was not infused. In the ASI treatment, i.v. infusion of artificial parotid saliva was initiated 1 h before feeding and continued for the entire 2 h feeding period. In the MI treatment, iso-osmotic mannitol solution was infused. The NI treatment showed that hematocrit and plasma total protein concentration were increased due to decreased circulating plasma volume brought about by feeding. In the ASI treatment, the fluid and $NaHCO_3$ that were lost from the blood because of a feeding-induced acceleration of saliva secretion was replenished with an intravenous infusion of artificial parotid saliva. This replenishment lessened the levels of suppression on both feeding and parotid saliva secretion. When only the lost fluid was replenished with an intravenous infusion of iso-osmotic mannitol solution in the MI treatment, the degree of feeding suppression was lessened but the level of saliva secretion suppression was not affected. These results indicate that the marked suppression of feed intake during the initial stages of dry forage feeding was caused by a feeding-induced hypovolemia while the suppression of saliva secretion was brought about by the loss of $NaHCO_3$ from the blood due to increased saliva secretion during the initial stages of feeding.
Background: Stress as a cause of mental health problems is known to be more prevalent in women than in men and has a negative effect on several aspects of physical health, such as the composition of blood and saliva. This study investigated the relationship of perceived stress with blood cell counts, saliva flow rate, and saliva factors. Methods: We recruited women in their 20s with a high prevalence of stress. Stress was evaluated using the Korean version of the perceived stress scale. Blood tests included white blood cell, hemoglobin, and platelet. We then examined the saliva flow rate and cariogenic bacteria level, acidity, occult blood, buffer capacity, leukocyte level, protein level, and ammonia level using rinse water with the SILL-HaⓇ saliva test system. Results: In a total of 70 participants, the average age was 21.64 years old, the average perceived stress score was 16.96±4.32, and high levels of stress were reported by 80% of the participants (n=56). The high-stress group had lower hemoglobin levels. In addition, the high-stress group showed a lower saliva flow rate than the low-stress group, and there was a difference in the salivary acidity and buffer capacity. The total perceived stress score showed a positive correlation with acidity and negative correlation with buffer capacity and the hemoglobin level. Conclusion: This study found that stress in female college students might affect the composition of blood and saliva. High levels of stress were positively correlated with the hemoglobin level, saliva flow rate, and acidity and negatively correlated with the buffer capacity.
Destruction of oral soft and hard tissues and resulting problems seriously affect the life quality of xerostomic patients. Although artificial saliva is the only regimen for xerostomic patients with totally abolished salivary glands, currently available artificial salivas give restricted satisfaction to patients. The purpose of this study was to contribute to the development of ideal artificial saliva through comparing viscosity and wettability between CMC solutions and human saliva. Commercially-available CMC is dissolved in simulated salivary buffer (SSB) and distilled deionized water (DDW). Various properties of human whole saliva, human glandular saliva, and a CMC-based saliva substitutes known as Salivart and Moi-Stir were compared with those of CMC solutions. Viscosity was measured with a cone-and-plate digital viscometer at six different shear rates, while wettability on acrylic resin and Co-Cr alloy was determined by the contact angle. The obtained results were as follows: 1. The viscosity of CMC solutions was proportional to CMC concentration, with 0.5% CMC solution displaying similar viscosity to stimulated whole saliva. Where as a decrease in contact angle was found with increasing CMC concentration. 2. The viscosity of human saliva was found to be inversely proportional to shear rate, a non-Newtonian (pseudoplastic) trait of biological fluids. The mean viscosity values at various shear rates increased as follows: stimulated parotid saliva, stimulated whole saliva, unstimulated whole saliva, stimulated submandibular-sublingual saliva. 3. Contact angles of human saliva on the tested solid phases were inversely correlated with viscosity, namely decreasing in the order stimulated parotid saliva, stimulated whole saliva, unstimulated whole saliva, stimulated submandibular-sublingual saliva. 4. Boiled CMC dissolved in SSB (CMC-SSB) had a lower viscosity than CMC-SSB (P < 0.01 at shear rate of $90s^{-1}$). 5. For human saliva, contact angles on acrylic resin were significantly lower than those on Co-Cr alloy (P < 0.01). 6. Comparing CMC solutions with human saliva, the contact angles between acrylic resin and human saliva solutions were significantly lower than those between acrylic resin and CMC solutions, including Salivart and Moi-Stir (P <0.01). The effectiveness of CMC solutions in terms of their rheological properties was objectively confirmed, indicating a vital role for CMC in the development of effective salivary substitutes.
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