Saliva Of Healthy Adults Research Articles

Increased Myeloperoxidase Concentrations in Saliva could Reflect Increased Body Mass and Oral Microinflammation.

Increased myeloperoxidase (MPO) levels in saliva are thought to reflect ongoing periodontal inflammation. Less clear is whether and to what extent salivary MPO is increased as a result of systemic inflammation. In the present study, we aimed to determine which demographic, anthropometric, biochemical, and dental parameters affect the level of MPO in whole mixed saliva in healthy adults with no apparent inflammatory lesions in the oral cavity. Thus, 113 individuals, aged 20-61 years (including 30.1% men and 23.9% smokers), were examined. In the univariate analysis, higher levels of MPO in saliva were found to be associated with age, an increased body mass index (BMI), higher levels of cytokines tumour necrosis factor-α and interleukin-6, as well as poorer oral hygiene, gingival status, and lower saliva flow. Multivariate logistic regression analysis determined that the main predictors of MPO concentration in saliva were BMI and stimulated saliva flow rate. Overall, an increase in MPO in saliva could be related to an increase in BMI, possibly as a result of subclinical chronic microinflammation, which also involves the gingiva.

Gustatory Function and Salivary Flow Rate in Healthy Adults.

In this study, we assessed gustatory function and whole-mouth saliva in healthy adults and investigated the association between gustatory function, salivary flow rate (SFR), and dry mouth. Descriptive clinical study. We recruited 413 participants who reported a normal sense of smell-taste, comprising 206 females and 207 males with an age range of 19 to 80 years. Taste scores were obtained using whole-mouth tests to evaluate the gustatory function, SFR was assessed using the spitting method, and dry mouth was measured with the visual analog scale. The total taste score (TTS) and SFR were significantly negatively correlated with age (rtaste =-0.522, P < .001; rSFR =-0.200, P < .001): TTS was significantly lower after the age of 50 and SFR decreased significantly after age 65. Women exhibited a significantly higher TTS than men (P < .001). However, there was no effect of sex on SFR. Dry mouth was negatively correlated with TTS (r=-0.223, P < .01) and SFR (r=-0.218, P < .01). Multiple regression analysis showed the independent variables of age, sex, and dry mouth predicted 34% of the variation in taste, while the independent variables of age and dry mouth predicted 6.1% of the variation in whole-mouth saliva. TTS was not significantly correlated with the SFR in healthy adults (r=0.094, P=.056). Age and dry mouth are important factors affecting gustatory function and whole-mouth saliva, with women exhibiting better taste sensitivity than men. Furthermore, there is no direct correlation between SFR and gustatory function in healthy adults. 3 Laryngoscope, 132:844-848, 2022.

Isolation and Characterisation of Probiotics for Antagonising the Cariogenic Bacterium Streptococcus mutans and Preventing Biofilm Formation.

Consumption of refined foods and beverages high in sugar make the teeth susceptible to the formation of biofilm and may lead to dental caries. The aim of the present study was to determine the ability of selected probiotics to inhibit growth and biofilm formation by the cariogenic bacterium Streptococcus mutans in vitro. Strains of latic acid bacteria (LAB) (n = 120) from the Bioresources Collection and Research Center (BCRC), saliva of healthy adults and infant stool were screened. The antimicrobial activity of LAB in vitro was evaluated by agar spot culture and co-culture of the S. mutans strains. Antagonistic substances in the spent culture suspensions (SCS) of LAB were precipitated by extraction with ammonium sulphate and chloroform to characterise the protein and lipophilic fractions. Results of co-culturing show that the SCS of the three LAB strains (Lactobacillus pentosus 13-1, 13-4 and L. crispatus BCRC 14618) subjected to heat treatment showed statistically significantly higher antimicrobial activity. Substances produced by L. pentosus 13-4 which have the potential to exhibit antimicrobial properties might be lipophilic proteins. Additionally, microtiter plate biofilm assays indicated that in vitro biofilm formation by S. mutans is strongly modulated by L. pentosus 13-4 and L. crispatus BCRC 14618. It can be inferred that the mechanism of reducing biofilm formation by these two LAB strains is associated with sucrose-dependent cell-cell adhesion and the gtfC level of glucosyltransferases in the biofilm. Therefore, it is suggested that L. pentosus 13-4 and L. crispatus BCRC 14618 may contribute to preventing dental caries, as they showed an inhibitory effect on the growth and biofilm formation of the cariogenic bacterium S. mutans in vitro.

A Randomized Single Dose Parallel Study on Enhancement of Nitric Oxide in Serum and Saliva with the Use of Natural Sports Supplement in Healthy Adults

ABSTRACT Sports supplements that stimulate the production of nitric oxide (NO) are widely promoted agents in the sports nutrition domain, and nitric oxide plays an important role to enhance the cardiovascular and physical fitness of the sports participants. The purpose of the study is to investigate whether oral intake of a sports nutritional supplement (Fitnox) is able to increase nitrate (NO3 −) and nitrite (NO2 −) levels in blood serum and saliva of healthy adults. Fitnox is a unique blend of Kaempferia parviflora methoxy flavones, pomegranate peel polyphenols, and Moringa oleifera leaf saponins. Twenty-four healthy male adults were equally divided and underwent the double-blind, placebo-controlled clinical trial with a single oral dose of sports nutrition formulation (250 mg capsules); blood and saliva samples were analyzed at different time intervals by high-performance liquid chromatography (HPLC). After administration of Fitnox (250 mg capsule as single dose), NO3 − and NO2 − levels in serum and saliva were found to be significantly higher (p <.05) than in the placebo group in 24 hours. Pharmacokinetic parameters such as the area under the plasma concentration-time curve extrapolated to infinity (AUC0-inf), AUC calculated to the last measured concentration (AUC0-t), maximum drug serum concentrations (Cmax), time of maximum concentration in serum observed (Tmax), and time required for the concentration of the drug to reach half of its original value (Thalf) were also statistically significant (p <.05) compared with the placebo. The results indicate that a single oral dose of Fitnox is able to increase the NO3 − and NO2 − levels considerably in the body relative to placebo for at least 12 hours. Therefore, Fitnox can improve the overall performance of sport participants and enhance physical endurance.

Pharmacokinetic study of amaranth extract in healthy humans: A randomized trial

ObjectiveNitric oxide (NO) is one of the most important signaling molecules produced within the body. Continuous generation of NO is essential for the integrity of the cardiovascular system. The aim of this study was to assess whether oral intake of a nitrate (NO3ˉ)-rich dietary supplement (amaranth extract) is able to increase NO3ˉ and nitrite (NO2ˉ) levels in blood plasma and saliva of healthy adults. MethodsIn the present study, bioavailability and pharmacokinetics of NO3ˉ and NO2ˉ from amaranth extract (2 g as single dose) was studied in 16 healthy individuals and compared with placebo in a crossover design. The NO3ˉ and NO2ˉ levels in plasma as well as saliva were measured up to 24 h. ResultsAfter administration of amaranth extract, the NO3ˉ levels in plasma as well as saliva were found to be significantly (P < 0.001) higher than in the placebo group. The NO2ˉ level in plasma was slightly higher (P < 0.05) in the amaranth group (test group) compared with that in the placebo group, whereas the saliva NO2ˉ level was significantly high (P < 0.001) in the amaranth extract–treated group than the placebo group. ConclusionsThese results clearly indicate that a single oral dose of amaranth extract is able to increase the NO3ˉ and NO2ˉ levels in the body for at least 8 h. The increase in NO3ˉ and NO2ˉ levels can help to improve the overall performance of people involved in vigorous physical activities or sports.

Search for varicella zoster virus DNA in saliva of healthy individuals aged 20–59 years

All neurological and ocular complications of varicella zoster virus (VZV) reactivation can occur without rash. Virological verification requires detection of VZV DNA or anti-VZV IgG antibody in cerebrospinal fluid (CSF), or anti-VZV IgM antibody in serum or CSF. If VZV were readily detected in other tissue in patients with neurological disease without rash and found to correlate with tests listed above, more invasive tests such as lumbar puncture might be obviated. Saliva is a potential source of VZV DNA. To study the potential diagnostic value of detecting VZV DNA in saliva from patients with neurological disease, saliva of healthy adults was searched for VZV DNA. A single saliva sample obtained by passive drool was centrifuged at 16,000g for 20 min. DNA was extracted from the supernatant and cell pellet and examined in triplicate for VZV DNA by real time PCR. A single random saliva sample from 80 healthy men and women aged 20-59 years revealed no VZV DNA (Table ), but was uniformly positive for cell (GAPdH) DNA. Because VZV DNA was not found in a random saliva sample from 80 individuals 20-59-year-old, a VZV-positive sample during neurologic disease may have potential significance. Further studies will determine whether VZV DNA in saliva correlates with VZV DNA or anti-VZV antibody in CSF in patients with neurological disease.

Comparison of Biomarkers in Blood and Saliva in Healthy Adults

Researchers measure biomarkers as a reflection of patient health status or intervention outcomes. While blood is generally regarded as the best body fluid for evaluation of systemic processes, substitution of saliva samples for blood would be less invasive and more convenient. The concentration of specific biomarkers may differ between blood and saliva. The objective of this study was to compare multiple biomarkers (27 cytokines) in plasma samples, passive drool saliva samples, and filter paper saliva samples in 50 healthy adults. Demographic data and three samples were obtained from each subject: saliva collected on filter paper over 1 minute, saliva collected by passive drool over 30 seconds, and venous blood (3 mL) collected by venipuncture. Cytokines were assayed using Bio-Rad multiplex suspension array technology. Descriptive statistics and pairwise correlations were used for data analysis. The sample was 52% male and 74% white. Mean age was 26 (range = 19–63 years, sd = 9.7). The most consistent and highest correlations were between the passive drool and filter paper saliva samples, although relationships were dependent on the specific biomarker. Correlations were not robust enough to support substitution of one collection method for another. There was little correlation between the plasma and passive drool saliva samples. Caution should be used in substituting saliva for blood, and relationships differ by biomarker.

RP-HPLC–ESI-MS evidenced that salivary cystatin B is detectable in adult human whole saliva mostly as S-modified derivatives: S-Glutathionyl, S-cysteinyl and S–S 2-mer

An HPLC-ESI-MS analysis of adult human whole saliva evidenced three protein masses (M average 11,487±2, 11,301±2 and 22,362±3Da) eluting in the 32.5-35.0min range. Treatment in reducing conditions allowed establishing that they are S-derivatives of N-terminal acetylated cystatin B, namely its S-glutathionyl, S-cysteinyl and S-S dimer. The identification was confirmed by high resolution HPLC-ESI-MS-MS experiments on the intact naturally occurring proteins and their tryptic digests. S-unmodified cystatin B is rarely detectable in whole saliva of healthy adults (5 subjects out of 65) and its percentage does not overcome approximately 20% of total cystatin B (11±9%). In the majority of subjects (60 out of 65) the mean percentages of the S-modified derivatives were S-glutathionyl 53±13%, S-cysteinyl 15±5%, S-S 2-mer 32±13%. Variations of the percentages of these S-modified derivatives of cystatin B could be indicative of oral oxidative stress. As we are aware, this is the first time that S-glutathionylation and S-cysteinylation were described as extensive PTM of a salivary protein and the first time that these PTMs were detected in naturally occurring cystatin B.

Detection of human herpesvirus 6 and 7 DNA in saliva from healthy adults from Rio de Janeiro, Brazil

In this study, we aimed to evaluate virus shedding in the saliva of healthy adults from the metropolitan region of the city of Rio de Janeiro, Brazil, in order to verify the prevalence of both human herpesviruses 6 and 7 (HHV-6, HHV-7). The studied group comprised 182 healthy individuals at Pedro Ernesto University Hospital, who were being seen for annual odontologic revisions. Saliva specimens were subjected to a multiplex polymerase chain reaction (PCR) to detect the presence of HHV-6A, HHV-6B and HHV-7. The total Roseolovirus DNA prevalence was 22.4%. The PCR detected a HHV-6 prevalence of 9.8%, with HHV-6A detected in 7.1% of the samples and HHV-6B in 2.7%. HHV-7 DNA was revealed in 12.6% of the studied cases. Multiple infections caused by HHV-6A and 7 were found in 2.1% of the samples. No statistical differences were observed regarding age, but for HHV-7 infection, an upward trend was observed in female patients. Compared to studies from other countries, low prevalence rates of herpesvirus DNA were detected in saliva from the healthy individuals in our sample. PCR methodology thus proved to be a useful tool for Roseolovirus detection and it is important to consider possible geographic and populations differences that could explain the comparatively low prevalence rates described here.

Detection of Homocysteine and C-Reactive Protein in the Saliva of Healthy Adults: Comparison with Blood Levels

Inflammation and cardiovascular disease are associated with elevated serum levels of C-Reactive Protein (CRP) and homocysteine. The presence of both molecules in saliva provides an opportunity for development of non-invasive assessments of disease risk. However, salivary CRP and homocysteine reference ranges and their correlation with serum levels are unknown. This study investigated if CRP and homocysteine could be routinely detected in the saliva of healthy adults and the relationship between salivary and blood levels. CRP and homocysteine concentrations were determined using ELISA and enzymatic assays respectively. Homocysteine was detected in only two saliva samples (n = 55). CRP was measurable in all saliva samples (range: 0.05 to 64.3 mug/L; median = 1.2 mug/L) and plasma samples (range: 0.14 to 31.1 mg/L; median = 2.0 mg/L). Regression analysis demonstrated no relationship between CRP concentration in saliva and plasma (R(2) = 0.001). Generalized linear models including variables such as saliva flow rate and time since eating or drinking also did not pass lack of fit testing. Therefore, a relationship between CRP concentration in saliva and blood could not be established in this group of subjects. More sensitive detection methods are needed to determine if a correlation between salivary and serum homocysteine levels exists.

Inter-individual variability of protein patterns in saliva of healthy adults

In order to document inter-individual variability in salivary protein patterns, unstimulated whole saliva was obtained from 12 subjects at 10 am and 3 pm of the same day. Saliva proteins were separated using two-dimensional gel electrophoresis, and semi-quantified using image analysis. One-way ANOVA was used to test the effects "time of sampling" and "subject". Data were further explored by multivariate analyses (PCA, hierarchical clustering). Spots of interest were identified by mass spectrometry (MALDI-TOF MS/MS and nanoLC ESI-IT MS/MS). A dataset of 509 spots matched in all gels was obtained. There was no diurnal statistical effect on salivary patterns while inter-individual variability was high with 47 spots differentially expressed between subjects (p<1%). Clustering of these spots revealed that subjects could be discriminated first based on several proteins participating to the non-specific immune response (cystatins, lipocalin 1, parotid-secretory protein and prolactin-induced protein). Independently, subjects were also differentiated by their level of proteins originating from serum and involved in the immune system (complement C3, transferrin, IgG2), as well as the relative abundance of enzymes involved in carbohydrates metabolism (amylase and glycolytic enzymes). Inter-individual variability should be accounted for when searching for salivary biomarkers or when studying in-mouth biochemical mechanisms.

Effect of Prophylactic Valacyclovir on the Presence of Human Herpesvirus DNA in Saliva of Healthy Individuals after Dental Treatment

Human herpesviruses (HHVs) are ubiquitous pathogens that intermittently reactivate from latency. Transmission is believed to be facilitated by their frequent appearance in saliva. This study sought to understand the factors that influence the appearance of these viruses in saliva by examining the prevalence, pattern, and quantity of all eight HHVs in saliva of immunocompetent adults with a history of recurrent oral herpes simplex virus (HSV) infections following dental treatment and antiviral therapy. Valacyclovir or matched placebo was given (2 g twice on the day of treatment and 1 g twice the following day) to 125 patients in a randomized, double-blind controlled trial. Saliva, collected on the day of dental treatment and 3 and 7 days later, was analyzed using real-time quantitative PCR. At all visits, HHVs coinfected saliva. Over the course of the week, the DNAs of HHV-6 and HHV-7 were detected significantly more often (97% to 99% of patients) than Epstein-Barr virus (EBV; 64.8%), HSV-1 (13.0%), HHV-8 (3.2%), cytomegalovirus (2.4%), HSV-2 (0%), and varicella-zoster virus (0%), irrespective of drug treatment (P < 0.002). Mean genome copy numbers were highest for HSV-1 and HHV-6. Dental treatment did not influence asymptomatic viral shedding patterns. However, valacyclovir treatment resulted in significantly fewer patients shedding EBV at both postoperative visits compared with placebo (P < 0.008). These results suggest that HHVs are simultaneously present in the saliva of healthy adults at levels that could facilitate transmission, and valacyclovir therapy decreases the prevalence of EBV in saliva but has little effect on HHV-6 and HHV-7.

Detection of penicillin-binding protein 2b gene alteration in Streptococcus mitis by polymerase chain reaction.

Three isolates of beta-lactam-resistant streptococci from the saliva of healthy adults were identified as Streptococcus mitis. Minimum inhibitory concentrations (MICs) were 2 to 4 micro g/ml for ampicillin (ABPC) and 64 to more than 128 micro g/ml for cefaclor (CCL). To determine the position of base alterations of the penicillin-binding protein 2b ( pbp2b) gene, upstream primers containing possible mutation points were designed, and used for polymerase chain reaction (PCR), together with a downstream primer. Alterations adjacent to the conserved motifs of the pbp2b gene were apparent. DNA sequencing data indicated replacements in deduced amino acid sequences in all resistant isolates: from threonine to alanine just after the serine-serine-asparagine (SSN) motif, and from alanine to glycine two residues downstream of the lysine-threonine-glycine (KTG) motif. These changes were the same as those in penicillin-resistant Streptococcus pneumoniae (PRSP), suggesting importance for the enzymatic activity of the protein. Thus, Beta-lactam susceptibility of S. mitis may be partially predicted by PCR using our primer set for pbp2b.

Monitoring of human herpesvirus-6 and -7 genomes in saliva samples of healthy adults by competitive quantitative PCR

Human herpesviruses-6 and -7 (HHV-6 and HHV-7) are thought to be transmitted during early infancy through saliva. However, the kinetics of the virus shedding in saliva of healthy adults, from whom children are assumed to acquire the viruses, is mostly unknown. This study was conducted to determine how many copies of the genome are secreted in saliva of healthy adults and to clarify the relationship between viral DNA load and virus isolation of HHV-6 and HHV-7. Competitive PCR was performed using primer sets in the U42 gene of each viral genome. In saliva samples from 29 healthy adults, HHV-6 and HHV-7 DNA was detected in 41.4% and 89.7%, respectively. The average copy number of the HHV-7 genome in the positive samples was higher than that of the HHV-6 genome. Follow-up studies of six seropositive individuals for 3 months showed that the amount of HHV-7 DNA was constant in each individual and that "high producers" and "low producers" could be distinguished. By contrast, the amount of HHV-6 DNA varied drastically over time in each individual. Although HHV-6 was never isolated from the saliva of any of the six individuals during the follow-up period, HHV-7 was isolated from each individual several times. The amount of HHV-7 DNA tended to be higher at the times when the virus was isolated than at the times when the virus was not isolated. These data demonstrate a striking contrast between HHV-6 and HHV-7 in the kinetics of genome and virus shedding.

Correlations between total protein, lysozyme, immunoglobulins, amylase, and albumin in stimulated whole saliva during daytime

The correlations between salivary proteins and the daytime variations are not known. The present study investigated the within-subject variation of correlations and concentrations between lysozyme, IgA, IgG, IgM, albumin, amylase, and total protein in stimulated whole saliva of healthy adults in the course of a 12-h period. After several practise sessions, unstimulated and stimulated whole saliva samples were collected five times daily (at 8 a.m., 11 a.m., 2 p.m., 5 p.m., and 8 p.m.) from 30 healthy university students. Flow rate and total protein concentration were used as covariates, and gender as a between-subject factor in the MANOVA analysis. After this adjustment, there was significant within-subject variation in salivary IgA (P < 0.001), albumin (P < 0.01), amylase (P < 0.05), and total protein (P < 0.001) concentrations. Total protein correlated significantly with amylase albumin and IgA through different samplings. In addition, IgG correlated with albumin and lysozyme in the course of 12 h. On the whole, the correlations between variables remained stable during repeated samplings. In addition, rankings of subjects for the variables tended to be maintained across different samplings (P < 0.001). However, the observed within-subject variations in salivary IgA, albumin, amylase, and total protein concentrations suggest that these proteins are subject to short-term variation.

Pathogenicity of the Streptococcus milleri group in pulmonary infections--effect on phagocytic killing by human polymorphonuclear neutrophils.

The pathogenicity of Streptococcus constellatus in pulmonary infections was investigated in mice in correlation with the phagocytic killing of the microorganisms by human polymorphonuclear neutrophils (PMNs). The histological inflammation score of the lungs in mice inoculated intratracheally with 10(8) cfu/mouse of the virulent strain S. constellatus RZYK001 obtained from bronchoalveolar fluid of a patient with pneumonia, was significantly higher than in mice inoculated with the avirulent strains S. constellatus RT4303 and RT6002 obtained from saliva of healthy adults (p < 0.001). The bactericidal activity of human PMNs against Staphylococcus aureus ATCC 25923 was not inhibited at all by the culture filtrate of the virulent strain, but the killing index against RZYK001 was 36.4% after 90-min incubation whereas that against RT4303 and RT6002 was 94.7% and 99.4%, respectively (p < 0.001). Therefore, these results suggest a possible pathogenic mechanism of S. constellatus apparently mediated by a structural component of the organisms so that a virulent strain is less likely to be killed than an avirulent one, predisposing it to survive in the infection sites.

Frequent isolation of human herpesvirus 7 from saliva

We obtained isolates of human herpesvirus 7 (HHV-7) from 6 of 8 healthy adults by culturing saliva with human umbilical cord blood lymphocytes. These isolates were identified as HHV-7 on the basis of comparisons of restriction endonuclease fragment profiles and hybridization with HHV-7 strain RK DNA. The isolates could be differentiated from HHV-7 strain RK and from each other by their restriction endonuclease fragment profiles. We confirm the finding of frequent isolation of HHV-7 from saliva of healthy adults and report the first dual isolation of human herpesvirus 6 (HHV-6) and HHV-7 from a single saliva specimen. We also describe an in situ hybridization assay that can distinguish between HHV-6 and HHV-7.

Human herpesvirus 7 is a constitutive inhabitant of adult human saliva.

We report the frequent isolation of human herpesvirus 7 from the saliva of healthy adults. Virus isolates recovered from different individuals exhibited minimal restriction enzyme polymorphism, which was mostly confined to heterogeneous (het) sequences in the genome. DNAs of isolates recovered from the same individual over a period of several months showed the same characteristic het fragments, indicating the stability of the het sequences upon virus replication and shedding in vivo. In contrast to the results of previous reports, human herpesvirus 6, the causative agent of roseola infantum, could not be isolated from the saliva specimens, raising questions regarding oral transmission of human herpesvirus 6 and human herpesvirus 7 to young children.

Transforming growth factor type alpha in normal human adult saliva

For the first time, transforming growth factor alpha (TGF-α) has been detected in the saliva of healthy adults. In saliva, the concentrations of TGF-α and epidermal growth factor (EGF) were determined by radioimmunoassay (RIA) to be 1–6 μg/1 saliva and 0.5–2 μg/1 saliva, respectively. The salivary TGF-α was partially purified about 1200-fold by an ethanol extraction, Bio-Gel P-30 molecular sieve chromatography and reverse-phase high-performance liquid chromatography (HPLC). At least three forms of TGF-α, with molecular weights of 6 kDa or less, were detected with molecular sieve gel filtration chromatography. A major chromatography peak, which constituted 20–30% of the extract, was a 5.6 kDa human TGF-α (hTGF-α) molecule by using RIA, radioreceptor assay (RRA), and mitogenic activity assay techniques. Reverse-phase HPLC purification further confirmed the identity of TGF-α. Our findings suggest, in contrast to what was earlier assumed, that TGF-α is present in normal adult saliva and we hypothesize that it may play a significant role in mediating proliferation and other functions of epithelial cells in the alimentary system.