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Item Binding of oral veillonella species to saliva-coated hydroxyapatite(1993) Wu, Sonya L.; Hughes, Christopher V.; Gregory, Richard L.; Sanders, Brian J.; Bowman, Dennis E.; Avery, David R.Veillonella spp. are found in high numbers in the mouth in dental plaque and on the mucosa. Veillonellae utilize lactic acid for their metabolic needs. A symbiotic relationship between Veillonellae and other oral bacteria, including a nutritional relationship with some streptococci, has been demonstrated both in vitro and in vivo. Thus, Veillonellae may protect the host from dental caries. Adherence is the initial step in bacterial colonization of oral surfaces. Recent evidence suggests that certain oral bacteria express molecules (adhesins) on their cell surface, which recognize receptors on other oral bacteria and/or in salivary pellicle. It has been previously demonstrated that Veillonella spp. bind avidly to Streptococcus. spp. found in subgingival plaque. The present study investigated the ability of V. atypica PK1910 to bind to saliva-coated hydroxyapatite (SHA), a model for adherence to the salivary pellicle. The results show that there was statistically significant enhanced binding of Veillonella atypica PK1910 to saliva-coated hydroxyapatite beads. (p< 0.05) Three classes of coaggregation-defective mutants of V. atypica PK1910 were tested for their ability to bind to SHA. Interestingly, they did not demonstrate any enhanced binding to saliva-coated hydroxyapatite beads. Heating of PK1910 did not effect binding to SHA. In contrast, protease treatment of the veillonella cell surface inactivated binding. Therefore, it appears that V. atypica PK1910, in addition to binding to oral Streptoccoccus spp. in dental plaque, may also colonize the tooth surface by binding directly to the salivary pellicle. It appears that a distinct heat stable protein may mediate this binding to SHA.Item Biochemical and genetic heterogeneity of the basic glycoproteins of parotid saliva(1971) Friedman, Robert D.Item A comparison of IgA antibody levels in caries-resistant and caries-susceptible children(1993) Rose, Paul Todd; Hughes, Christopher V.; Avery, David R.; Sanders, Brian J.; Branca, Ronald A.; Gregory, Richard L.Secretory immunity is believed to play a role in natural resistance to dental caries. Although dental caries has dramatically decreased in children in the United States, there remains a population of caries-susceptible children even in fluoridated communities. Previous studies have shown a positive correlation between salivary immunoglobulin A (sIgA) antibody levels to Streptococcus mutans and caries resistance in adults. In the present study, an enzyme-linked immunosorbent assay (ELISA) was used to compare IgA antibody levels to S. mutans in saliva from 20 caries susceptible (DMFS greater than 5) and 20 caries-resistant (DMFS less than or equal to 1) children (ages 7-11). All subjects resided in fluoridated communities. Salivary S. mutans numbers were significantly higher (p ≤ 0.05) in the caries susceptible (31.2 percent of total streptococci) group than in the caries resistant (1.6 percent of total streptococci) group. Whole saliva from caries-resistant children had significantly higher (p = 0.05) levels of IgA antibodies to S. mutans than saliva from caries-susceptible children. However, whole saliva from caries-resistant children had similar levels of IgA1 or IgA2 antibodies against S. mutans to saliva from caries-susceptible children. These results suggest that IgA antibody to S. mutans may play a role in natural protection from dental caries in children and confirm previous reports indicating a role for salivary IgA antibodies to S. mutans in mediation of caries.Item COVID-19 and saliva: A primer for dental health care professionals(Wiley, 2020-08-23) Srinivasan, Mythily; Thyvalikakath, Thankam P.; Cook, Blaine N.; Zero, Domenick T.; Oral Pathology, Medicine and Radiology, School of DentistryTo contain the COVID‐19 pandemic, it is essential to find methods that can be used by a wide range of health care professionals to identify the virus. The less potential contagious nature of the collection process, the ease of collection and the convenience of frequent collection for real‐time monitoring makes saliva an excellent specimen for home‐based collection for epidemiological investigations. With respect to COVID‐19, the use of saliva offers the added advantages of greater sensitivity and potential for detection at an early stage of infection. However, the advantages from a diagnostic perspective also reflect the potential risk to dental professionals from saliva from infected patients. Although not validated in COVID‐19 patients, but by extension from studies of SARS‐CoV‐1 studies, it is suggested that using antimicrobial mouthrinses such as chlorhexidine, hydrogen peroxide or sodium hypochlorite solutions could reduce the viral load in saliva droplets and reduce the risk of direct transmission. Because large saliva droplets could deposit on inanimate surfaces, changing the personal protective equipment including the clinical gown, gloves, masks, protective eye wear and face shield between patients, as well as decontamination of the work surfaces in the clinic, could reduce the risk of indirect contact transmission.Item Critical Comparison of Total Vaporization- Solid Phase Microextraction vs Headspace- Solid Phase Microextraction(2021-05) Train, Alexandra; Goodpaster, John; Manicke, Nicholas; Picard, ChristineSolid Phase Microextraction (SPME) is a popular sampling technique that can be paired with Gas Chromatography/Mass Spectrometry (GC-MS). SPME-GC-MS is used in forensic chemistry due to its simplification of the sample preparation process. Headspace-Solid Phase Microextraction (HS-SPME) is a technique where the sample is heated to generate volatiles in the headspace of the vial. A SPME fiber is then inserted into the vial and the compounds in the headspace will bind to the fiber. Total Vaporization- Solid Phase Microextraction (TV-SPME) is a technique that is derived from the HS-SPME technique. In Chapter 1, the critical comparison of HS-SPME and TV-SPME is discussed. Samples including marijuana, essential oils, and CBD oil were utilized to compare the two techniques. The compounds of interest in marijuana are the three main cannabinoids: cannabinol (CBN), cannabidiol (CBD), and tetrahydrocannabinol (THC). The sample preparation and GC-MS parameters were kept the same for all samples to determine which SPME technique works best for these sample types and yielded the greatest sensitivity. It was found that HS-SPME shows greater sensitivity with CBN and equivalent sensitivity with essential oils, THC and CBD. In Chapter 2, the detection of synthetic cannabinoids utilizing liquid-liquid injection as well as HS-SPME and TV-SPME is discussed. The detection of these compounds is important because this type of drug has become more prevalent in the United States because they can be chemically altered slightly so they still have the effects of a drug but can evade drug legislation. The detection of synthetic cannabinoids using liquid injection was found to be successful but detection using HS-SPME and TV-SPME was found to be unsuccessful. In Chapter 3, the analyses of real and artificial saliva utilizing HS-SPME and TV-SPME is discussed. Determining the compounds present in real saliva and artificial saliva will be of importance for future research into determining if the presence of drugs in saliva can be analyzed with these techniques. The analyses of real and artificial saliva were found to be successful using HS-SPME, without derivatization, and TV-SPME, with and without derivatization. Many of the compounds present in the real saliva were detected and were confirmed to be compounds regularly found in saliva by other scientific literature.Item Differential profiles of soluble and cellular toll like receptor (TLR)-2 and 4 in chronic periodontitis(PLOS, 2018-12-20) AlQallaf, Hawra; Hamada, Yusuke; Blanchard, Steven; Shin, Daniel; Gregory, Richard; Srinivasan, Mythily; Periodontology, School of DentistryChronic periodontitis is a common inflammatory disease initiated by a complex microbial biofilm and mediated by the host response causing destruction of the supporting tissues of the teeth. Host recognition of pathogens is mediated by toll-like receptors (TLRs) that bind conserved molecular patterns shared by large groups of microorganisms. The oral epithelial cells respond to most periodontopathic bacteria via TLR-2 and TLR-4. In addition to the membrane-associated receptors, soluble forms of TLR-2 (sTLR-2) and TLR-4 (sTLR-4) have been identified and are thought to play a regulatory role by binding microbial ligands. sTLR-2 has been shown to arise from ectodomain shedding of the extracellular domain of the membrane receptor and sTLR-4 is thought to be an alternate spliced form. Many studies have previously reported the presence of elevated numbers of viable exfoliated epithelial cells in the saliva of patients with chronic periodontitis. The objective of this study was to investigate the potential value of salivary sTLR-2 and sTLR-4 together with the paired epithelial cell-associated TLR-2/4 mRNA as diagnostic markers for chronic periodontitis. Unstimulated whole saliva was collected after obtaining informed consent from 40 individuals with either periodontitis or gingivitis. The sTLR-2 and sTLR4 in saliva was measured by enzyme-linked immunosorbent assay. The TLR-2 and TLR-4 transcript in the epithelial cells in saliva was measured by real time polymerase chain reaction. While levels of sTLR-2 exhibited an inverse correlation, sTLR-4 positively correlated with clinical parameters in the gingivitis cohort. Interestingly, both correlations were lost in the periodontitis cohort indicating a dysregulated host response. On the other hand, while the sTLR-2 and the paired epithelial cell associated TLR-2 mRNA exhibited a direct correlation (r2 = 0.62), that of sTLR4 and TLR-4 mRNA exhibited an inverse correlation (r2 = 0.53) in the periodontitis cohort. Collectively, assessments of salivary sTLR2 and sTLR4 together with the respective transcripts in the epithelial cells could provide clinically relevant markers of disease progression from gingivitis to periodontitis.Item Distinct Salivary Biomarker Profile in Chronic Periodontitis(Office of the Vice Chancellor for Research, 2012-04-13) Prakasam, S.; Srinivasan, MythilyBackground: Saliva has potential to diagnose chronic periodontitis (CP). Changes in tissue-expression of pattern-recognition-receptors (PRRs), which recognize periodontal-pathogens, correlate with CP. It follows that PRRs-expression in nucleated-cells (NCs) shed in saliva and soluble-PRRs may differentiate CP from health. Additionally, cytokines in gingival cervical fluid (GCF) correlate with worsening CP, which may be reflected in saliva. One significant test for biomarkers is changes in response to treatment. Objectives: Comparison of CP salivary-biomarkers profile with health and to study treatment effects of scaling and root planning (SRP). Methods: Unstimulated whole saliva (UWS) collection and recording of routine clinical periodontal parameters was done for two groups (n=16): healthy (H) (minimal clinical loss of attachment (CAL) and clinical inflammation) and CP (≥30% sites with ≥4mm CAL). UWS was collected at 3 different time points: before, 1-week and 6-weeks after SRP from the CP group. NCs and clarified saliva (CS) were separated from UWS. Messenger RNA was extracted from NCs and TLR-2 expression was quantitated through real-time-PCR. CS depleted of immunoglobulin and amylase to prevent large molecule interferences and diluted to 1 μg/ml of salivary-protein in PBS, normalize for variations in liquid volume, was used to quantify biomarkers through ELISA. Statistical significance between H- and CP-groups biomarkers was determined through Mann-Whitney ‘U' test and one tailed paired ‘t' test. Results: Statistically significant differences were noted for clinical profiles of H- and CP-groups and for changes after SRP within CP-group. Salivary sTLR-2, IL-17 and IL-10, were significantly higher, and sCD14, IL-6, IL-4 and TLR-2 mRNA were significantly lower in H compared to CP. In CP, salivary sTLR-2 and IL10 increased significantly at 1- and 6-weeks after SRP, whilst IL-4 decreased significantly at 6-weeks. Conclusions: Salivary biomarkers profiles are distinct between health and CP as well as before and after SRP treatment. sTLR-2, IL-10 and IL-4 may serve as short-term biomarkers for monitoring response to SRP. sCD14, TLR2-mRNA and other cytokines need exploration as long-term response biomarkers. Depletion of amylase and immunoglobulin, and normalization for total salivary protein may be important in biomarkers quantification.Item Effects of cardiac glycosides on the composition of whole-mixed human saliva(1978) McDonald, John S., 1947-; Shafer, William G.; Barton, Paul; Bixler, David; El-Kafrawy, Abdel Hady, 1935-; Standish, S. Miles, 1923-2003Electrolyte levels were measured in whole-mixed human saliva collected from cardiology out-patients, to investigate any salivary electrolyte changes occurring in such patients after digitalization. Several recent reports have indicated that clinical symptoms of digitalis intoxication were associated with increased saliva concentrations of K+ and/or Ca++. Because salivary glands contain a highly active Na+, K+-ATPase it seemed logical that these and other salivary electrolytes might be predictably affected by the circulating levels of digitalis. Patients receiving digitoxin (Dtxn) had a higher concentration of Salivary K+ and Ca++ (25.8 ± 2.2 and 2.2 ± 0.2 meq/l, respectively), than the controls not receiving cardiac glycosides (20.1 ± 1.4 and 2.0 ± 0.1 meq/1, respectively). A similar pattern was not found for patients receiving digoxin (Dxn), although the mean Ca++ concentration for females in this group was significantly elevated (control: 1.7 ± 0.2; Dxn: 2.4 ± 0.2 meq/1). The mean serum concentration (ng/ml) of Dtxn was 20.3 ± 1. 9; of Dxn, 1.4 ± 0.2. No change was found in P04, and protein concentrations, or in salivary flow rates between control and experimental groups. The results suggest that salivary electrolyte changes occur after digitalization, but that these changes do not adequately reflect the serum level of digitalis in individual patients. This study was supported in part by PHS 80l-RR5312.Item Expensive Egos: Narcissistic Males Have Higher Cortisol(PLOS, 2012-01-23) Reinhard, David A.; Konrath, Sara H.; Lopez, William D.; Cameron, Heather G.Background Narcissism is characterized by grandiosity, low empathy, and entitlement. There has been limited research regarding the hormonal correlates of narcissism, despite the potential health implications. This study examined the role of participant narcissism and sex on basal cortisol concentrations in an undergraduate population. Methods and Findings Participants were 106 undergraduate students (79 females, 27 males, mean age 20.1 years) from one Midwestern and one Southwestern American university. Narcissism was assessed using the Narcissistic Personality Inventory, and basal cortisol concentrations were collected from saliva samples in a laboratory setting. Regression analyses examined the effect of narcissism and sex on cortisol (log). There were no sex differences in basal cortisol, F (1,97) = .20, p = .65, and narcissism scores, F (1,97) = .00, p = .99. Stepwise linear regression models of sex and narcissism and their interaction predicting cortisol concentrations showed no main effects when including covariates, but a significant interaction, β = .27, p = .04. Narcissism was not related to cortisol in females, but significantly predicted cortisol in males. Examining the effect of unhealthy versus healthy narcissism on cortisol found that unhealthy narcissism was marginally related to cortisol in females, β = .27, p = .06, but significantly predicted higher basal cortisol in males, β = .72, p = .01, even when controlling for potential confounds. No relationship was found between sex, narcissism, or their interaction on self-reported stress. Conclusions Our findings suggest that the HPA axis is chronically activated in males with unhealthy narcissism. This constant activation of the HPA axis may have important health implications.Item Human Saliva: A study of the rate of flow and viscosity and its relationship to dental caries(1950) McDonald, Ralph E.