• Title, Summary, Keyword: saliva

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The Effects of Storage of Human Saliva on DNA Isolation and Stability (인체타액의 보관이 DNA 분리와 안정도에 미치는 영향)

  • Kim, Yong-Woo;Kim, Young-Ku
    • Journal of Oral Medicine and Pain
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    • v.31 no.1
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    • pp.1-16
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    • 2006
  • 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.

Changes of Halitosis during the Menstrual Cycle (월경주기와 구취의 변화에 관한 연구)

  • Kim, In-Jung;Choi, Jong-Hoon;Kim, Chong-Youl
    • Journal of Oral Medicine and Pain
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    • v.26 no.1
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    • pp.17-25
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    • 2001
  • 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).

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Bacterial Diversity in the Human Saliva from Different Ages

  • Kang, Jung-Gyu;Kim, Seong-Hwan;Ahn, Tae-Young
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.572-576
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    • 2006
  • 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.

Influences of Saliva Substitutes on Salivary Enzymatic Activity (타액대체제가 타액 효소 활성에 미치는 영향)

  • Kho, Hong-Seop;Lee, Sung-Woo
    • Journal of Oral Medicine and Pain
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    • v.34 no.3
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    • pp.227-235
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    • 2009
  • 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.

Physiological Factors Depressing Feed Intake and Saliva Secretion in Goats Fed on Dry Forage

  • Sunagawa, K.;Ooshiro, T.;Nakamura, N.;Ishii, Y.;Nagamine, I.;Shinjo, A
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.1
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    • pp.60-69
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    • 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.

Viscosity and Wettability of Carboxymethylcellulose(CMC) solutions and Artificial Saliva (Carboxymethylcellulose(CMC) 용액과 인공 타액의 점도와 습윤성)

  • Park, Moon-Soo;Kim, Young-Jun
    • Journal of Oral Medicine and Pain
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    • v.32 no.4
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    • pp.365-373
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    • 2007
  • 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.

AN ELECTROCHEMICAL STUDY BY USING A POTENTIOSTAT ON THE CORROSION OF AMALGAMS IN SALIVA (Potentiostat를 이용한 타액에서의 아말감부식에 대한 전기화학적 연구)

  • Son, Yoon-Hee;Um, Chung-Moon
    • Restorative Dentistry and Endodontics
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    • v.19 no.2
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    • pp.534-548
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    • 1994
  • The purpose of this study is to observe the corrosion characteristics of seven dental amalgams (CAULK FINE CUT, CAULK SPHERICAL, OPTALLOY II, DISPERSALLOY, HI VERALOY, TYTIN, VALIANT) through the anodic polarization curve obtained by using a potentiostat. After each amalgam alloy and Hg being triturated, the triturated mass was inserted into the cylindrical metal mold, and condensed by hydrolic pressure(160 kg/$cm^2$). Each specimen was removed from the metal mold. 24 hours after condensation, specimens were polished with the emery paper and stored at room temperature for 1 week. The anodic polarization curves were employed to compare the corrosion behaviours of the amalgam in 0.9% saline solution, Fusayama's artificial saliva, and stimulated parotid saliva at $37^{\circ}C$ with 3-electrode potentiostat. After the immersion of specimen in electrolyte for 1 hour, the potential scan was begun. The potential scan range was. -1700m V ~ + 400m V(vs. S. C. E) in the working electrode and the scan rate was 50m V /sec. The results were as follows, 1. The corrosion potential, the potential of anodic current peak, and transpassive potential in the stimulated parotid saliva shifted to more anodic direction than those in saline solution, and the current density in the stimulated parotid saliva was lower than that in saline solution. Those in Fusayama's artificial saliva was similar to those in stimulated parotid saliva. 2. The anodic polarization profiles in Fusayama's artificial saliva and stimulated parotid saliva indicated a region of slow slope current density, which is extending from the corrosion potential to the potential of anodic current peak, but that in 0.9% saline solution indicated no region of slow slope. 3. The corrosion potentials for CAULK FINE CUT, CAULK SPHERICAL, and OPT ALLOY II had the similarity in 0.9% saline solution, Fusayama's artificial saliva and stimulated parotid saliva, but those for high coper amalgam and VALIANT had no similarity. 4. The current density for TYTIN amalgam in stimulated parotid saliva was the lowest among the others. 5. As for current density, there was no significant difference between palladium enriched VALINAT and other high copper amalgams.

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The Effect of Pilocarpine-containing Chewing Gum on Anti-microbial Components in Whole Saliva of Xerostomic Patients (구강건조증 환자에서 필로카핀 함유 껌 사용에 따른 전타액내 항균성분의 변화)

  • Park, Moon-Soo;Lee, Sung-Woo;Chung, Sung-Chang;Kim, Young-Ku;Yum, Kwang-Won
    • Journal of Oral Medicine and Pain
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    • v.24 no.4
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    • pp.347-359
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    • 1999
  • The purpose of this study was to investigate the effect of pilocarpine containing chewing gum on anti-microbial components in whole saliva of xerostomic patients, The objective xerostomic patients were instructed to use 5mg-pilocarpine containing chewing gum for 20minutes three times per day, and the author measured the flow rates of unstimulated whole saliva and stimulated whole saliva at the beginning the treatment, 1,2,3, and 4 weeks after. The concentration and flow rate of anti-microbial components in whole saliva were quantitated by enzyme-linked immunosorbent assay(ELISA). The obtained results were as follows: 1. There were significant increase in the unstimulated and stimulated whole salivary flow rate after using pilocarpine-containing chewing gum in xerostomic patients. 2. The concentrations of IgA in the unstimulated and stimulated whole saliva showed increasing pattern but, no significant changes, arid the flow rates of IgA in the unstimulated and stimulated whole saliva showed significant increase after using pilocarpine-containing chewing gum in xerostomic patients. 3. The concentrations of IgM in the unstimulated and stimulated whole saliva showed increasing pattern but, no significant changes, and the flow rates of IgM in the unstimulated and stimulated whole saliva showed significant increase after using pilocarpine-containing chewing gum in xerostomic patients. 4. The concentrations of lactoferrin in the unstimulated and stimulated whole saliva showed no significant changes, and the flow rates of lactoferrin in the unstimulated and stimulated whole saliva showed significant increase after using pilocarpine-containing chewing gum in xerostomic patients. 5. The concentrations of lysozyme in the unstimulated and stimulated whole saliva showed no significant changes, and the flow rates of lysozyme in the unstimulated whole saliva showed significant increase, but in stimulated whole saliva showed no significant changes after using pilocarpine-containing chewing gum in xerostomic patients.

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EFFECT OF FLUORIDE AND CALCIUM ON ENAMEL REMINERALIZATION IN VITRO (불소와 칼슘의 법랑질 재광화 효과에 대한 생체외 연구)

  • Lee, Kwang-Hee
    • THE JOURNAL OF THE KOREAN ACADEMY OF PEDTATRIC DENTISTRY
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    • v.31 no.4
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    • pp.624-629
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    • 2004
  • The purpose of study was to observe the effect of fluoride and calcium on enamel remineralizaton in vitro. Human premolar enamel specimens were prepared by demineralization in $0.1{\sim}l.0%$ citric acid for 60 minutes. They were remineralized for 6 hours in one of the 1311owing solutions : (1) artificial saliva, (2) artificial saliva with 100ppmF, (3) artificial saliva with 1000ppmF, (4) artificial saliva with 1000ppmCa, and (5) artificial saliva with 100ppmF and 1000ppmCa. No significant remineralization was occurred in artificial saliva and artificial saliva with 100ppmF. Significant remineralization was observed in artificial saliva with 1000ppmF at 3 hours, and in artificial saliva with 1000ppmCa and artificial saliva with 100ppmF and 1000ppmCa at 3 and 6 hours(P<0.05). The remineralization effect of artificial saliva with 100ppmF and 1000ppmCa was greater than that of artificial saliva or artificial saliva with 100ppmF. Addition of F to 100ppm or 1000ppm, addition of Ca to 1000ppm, and increasing the concentration of F from 100ppm to 1000ppm did not significantly increase the remineralization.

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A Study on pH, Viscosity, Microorganisms and Immunoglobulin A of the Saliva (타액의 수소이온 농도, 점도, 세균 및 면역글로블린 A에 관한 연구)

  • Jin-Woo Sohn;Sung-Woo Lee
    • Journal of Oral Medicine and Pain
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    • v.17 no.2
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    • pp.87-97
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    • 1992
  • The purpose of this study is to investigate the age-and sex-related changes in the pH of resting saliva, viscosity, microorganisms and immunoglobulin A of stimulated whole saliva, and to investigate their correlations. The 120 healthy subjects were included in this study and the author used cone-and plate digital viscometer for viscosity, MSB agar for Streptococcus mutans, SL Rogosa agar for lactobacilli, and single radial immunodiffusion technique for immunoglobulinA. The obtained results were as follows : 1. There was no significant difference in pH, viscosity, Streptococcus mutans lactobacilli and immunoglobulin A of the saliva between males and females. 2. The viscosity values of stimulated whole saliva showed the increasing pattern with aging. 3. DMFS (or dmfs) rate was not correlated with pH, viscosity, Streptococcus mutans, lactobacilli and immunoglobulin A of the saliva. 4. There was a significant difference in the concentration of immunoglobulin A between the group under 10 and groups above 10. 5. The viscosity values of stimulated whole saliva showed the increasing pattern with decreasing of the number of Streptococcus mutans.

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