Human saliva as a tool to investigate intimate partner violence

Human saliva mirrors body’s health and well-being and many of the biomolecules present in blood or urine can also be found in salivary secretions. However, biomolecular concentrations in saliva are usually one tenth to one thousandth of the levels in blood (Pfaffe et al., 2011). Sensitive detection technology platforms are therefore required to detect biomolecules in saliva. Another road block to the advancement of salivary diagnostics is the lack of information related to healthy state saliva vs. a diseased saliva, baseline levels and reference ranges and diurnal variations. Saliva has numerous advantages over blood or urine as a diagnostic fluid: (a) the non-invasive nature of sample collection and the simple, safe, painless and cost-effective methods to collect it; (b) unskilled personnel can collect saliva samples at multiple time points; and (c) the total protein concentration is approximately a quarter of that is present in plasma, which makes it easier to investigate low abundance proteins (Pfaffe et al., 2011). Currently, saliva assays are routinely used to determine, diseases such as HIV, drugs and substances of abuse to provide information on exposure and give qualitative information on the type of illicit drug used (Kintz et al., 2009), cortisol levels for diagnosing Cushing’s syndrome (Doi et al., 2008), and use for biomonitoring of exposure to chemicals (Caporossi et al., 2010) to measure hormones (Gröschl, 2009)....

Measurement of melatonin in body fluids: Standards, protocols and procedures

Background and Purpose: The circadian rhythm of melatonin in saliva or plasma, or of the melatonin metabolite 6-sulfatoxymelatonin (a6MTs) in urine, is a defining feature of suprachiasmatic nucleus (SCN) function, the body's endogenous oscillatory pacemaker. The primary objective of this review is to ascertain the clinical benefits and limitations of current methodologies employed for detection and quantification of melatonin in biological fluids and tissues. Data Identification: A search of the English-language literature (Medline) and a systematic review of published articles were carried out. Study Selection: Articles that specified both the methodology for quantifying melatonin and indicated the clinical purpose were chosen for inclusion in the review. Data Extraction: The authors critically evaluated the methodological issues associated with various tools and techniques (e.g. standards, protocols, and procedures). Results of Data Synthesis: Melatonin measurements are useful for evaluating problems related to the onset or offset of sleep and for assessing phase delays or advances of rhythms in entrained individuals. They have also become an important tool for psychiatric diagnosis, their use being recommended for phase typing in patients suffering from sleep and mood disorders. Additionally, there has been a continuous interest in the use of melatonin as a marker for neoplasms of the pineal region. Melatonin decreases such as found with aging are or post pinealectomy can cause alterations in the sleep/wake cycle. The development of sensitive and selective methods for the precise detection of melatonin in tissues and fluids has increasingly been shown to have direct relevance for clinical decision making. Conclusions: Due to melatonin's low concentration, as well as the coexistence of numerous other compounds in the blood, the routine determination of melatonin has been an analytical challenge. The available evidence indicates however that these challenges can be overcome and consequently that evaluation of melatonin's presence and activity can be an accessible and useful tool for clinical diagnosis. © Springer-Verlag 2010.

One-step homogeneous C-reactive protein assay for saliva

Background: Cardiovascular disease is the leading cause of death in the world. Human C-reactive protein (CRP) has been used in the risk assessment of coronary events. Human saliva mirrors the body's health and well-being and is non-invasive, easy to collect and ideal for third world countries as well as for large patient screening. The aim was to establish a saliva CRP reference range and to demonstrate the clinical utility of salivary CRP levels in assessing the coronary events in a primary health care setting. Methods: We have used a homogeneous bead based assay to detect CRP levels in human saliva. We have developed a rapid 15 min (vs 90 min), sequential, one-step assay to detect CRP in saliva. Saliva was collected from healthy volunteers (n = 55, ages 20-70 years) as well as from cardiac patients (n = 28, ages 43-86 years). Results: The assay incubation time was optimised from 90 min to 15 mm and generated a positive correlation (n = 29, range 10-2189 pg/mL, r2 = 0.94; Passing Bablok slope 0.885. Intercept 0, p>0.10), meaning we could decrease the incubation time and produce equivalent results with confidence. The mean CRP level in the saliva of healthy human volunteers was 285 pg/mL and in cardiac patients was 1680 pg/mL (p<0.01). Analysis of CRP concentrations in paired serum and saliva samples from cardiac patients gave a positive correlation (r2 = 0.84, p<0.001) and the salivary CRP concentration capable of distinguishing healthy from diseased patients. Conclusions: The results suggest that this minimally invasive, rapid and sensitive assay will be useful in large patient screening studies for risk assessment of coronary events. (C) 2011 Elsevier B.V. All rights reserved.

Pathological changes at the target tissue level in Sjögren's syndrome and their effect on the exocrine function of the salivary glands

Sjögren s syndrome (SS) is a common autoimmune disease affecting the lacrimal and salivary glands. SS is characterized by a considerable female predominance and a late age of onset, commonly at the time of adreno- and menopause. The levels of the androgen prohormone dehydroepiandrosterone-sulphate (DHEA-S) in the serum are lower in patients with SS than in age- and sex-matched healthy control subjects. The eventual systemic effects of low androgen levels in SS are not currently well understood. Basement membranes (BM) are specialized layers of extracellular matrix and are composed of laminin (LM) and type IV collagen matrix networks. BMs deliver messages to epithelial cells via cellular LM-receptors including integrins (Int) and Lutheran blood group antigen (Lu). The composition of BMs and distribution of LM-receptors in labial salivary glands (LSGs) of normal healthy controls and patients with SS was assessed. LMs have complex and highly regulated distribution in LSGs. LMs seem to have specific tasks in the dynamic regulation of acinar cell function. LM-111 is important for the normal acinar cell differentiation and its expression is diminished in SS. Also LM-211 and -411 seem to have some acinar specific functional tasks in LSGs. LM-311, -332 and -511 seem to have more general structure maintaining and supporting roles in LSGs and are relatively intact also in SS. Ints α3β1, α6β1, α6β4 and Lu seem to supply structural basis for the firm attachment of epithelial cells to the BM in LSGs. The expression of Ints α1β1 and α2β1 differed clearly from other LM-receptors in that they were found almost exclusively around the acini and intercalated duct cells in salivons suggesting some type of acinar cell compartment-specific or dominant function. Expression of these integrins was lower in SS compared to healthy controls suggesting that the LM-111 and -211-to-Int α1β1 and α2β1 interactions are defective in SS and are crucial to the maintenance of the acini in LSGs. DHEA/DHEA-S concentration in serum and locally in saliva of patients with SS seems to have effects on the salivary glands. These effects were first detected using the androgen-dependent CRISP-3 protein, the production and secretion of which were clearly diminished in SS. This might be due to the impaired function of the intracrine DHEA prohormone metabolizing machinery, which fails to successfully convert DHEA into its active metabolites in LSGs. The progenitor epithelial cells from the intercalated ductal area of LSGs migrate to the acinar compartment and then undergo a phenotype change into secretory acinar cells. This migration and phenotype change seem to be regulated by the LM-111-to-Int α1β1/Int α2β1 interactions. Lack of these interactions could be one factor limiting the normal remodelling process. Androgens are effective stimulators of Int α1β1 and α2β1 expression in physiologic concentrations. Addition of DHEA to the culture medium had effective stimulating effect on the Int α1β1 and α2β1 expression and its effect may be deficient in the LSGs of patients with SS.

Monitoring of saliva pH using miniaturized sensing tools as an indicator of oral health

The monitoring of oral disease is important, not alone for oral health, but for the detection and prevention of
systemic disease. The link between oral health and systemic disease is the focus of many studies, with
indications emerging of a causal link [1]. For disease diagnostics, blood has typically been the fluid of choice
for analysis, the retrieval of which is invasive and therefore unsuitable for wearable technology. Analysis of
saliva, however, is less invasive than that of blood, requires little or no pre-treatment and is abundantly
available. A strong correlation has been found between the analytes of blood and saliva [2] with saliva
containing biomarkers for diseases such as diabetes, oral cancer and cardiovascular disease. The development of
an implantable multi-parametric wireless sensor, to monitor both salivary analytes and changes in gingival
temperature, is the aim of this research project.
The aim of our current study is to detect changes in salivary pH, using a gold electrode with a pHsensitive
iridium oxide layer, and an Ion Sensitive Field Effect Transistor probe. Characterisation studies were
carried out in artificial saliva (AS). A salivary pH of between 4.5pH-7.5pH [3], and gingival temperature
between 35°C-38°C [4], were identified as the target range of interest for the human oral environment. Sensor
measurements were recorded in solutions of varying pH and temperature. An ISFET probe was then implanted
into a prototype denture and characterised in AS. This study demonstrates the suitability of ISFET and gold
electrode pH sensors for incorporation into implantable oral sensors.
[1] G. Taylor and W. Borgnakke, “Periodontal disease: associations with diabetes, glycemic control and
complications,” Oral Dis., vol. 14, no. 3, pp. 191–203, Apr. 2008.
[2] E. Tékus, M. Kaj, E. Szabó, N. L. Szénási, I. Kerepesi, M. Figler, R. Gábriel, and M. Wilhelm,
“Comparison of blood and saliva lactate level after maximum intensity exercise,” Acta Biol. Hung., vol. 63
Suppl 1, pp. 89–98, 2012.
[3] S. Naveen, M. L. Asha, G. Shubha, A. Bajoria, and A. Jose, “Salivary Flow Rate, pH and Buffering
Capacity in Pregnant and Non Pregnant Women - A Comparative Study,” JMED Res., pp. 1–8, Feb. 2014.
[4] A. F. Holthuis and F. S. Chebib, “Observations on temperature and temperature patterns of the gingiva. I.
The effect of arch, region and health,” J. Periodontol., vol. 54, no. 10, pp. 624–628, Oct. 1983

Myeloperoxidase activity is increased in gingival crevicular fluid and whole saliva after fixed orthodontic appliance activation

Introduction: Orthodontic tooth movement uses mechanical forces that result in inflammation in the first days. Myeloperoxidase (MPO) is an enzyme found in polymorphonuclear neutrophil (PMN) granules, and it is used to estimate the number of PMN granules in tissues. So far, MPO has not been used to study the inflammatory alterations after the application of orthodontic tooth movement forces. The aim of this study was to determine MPO activity in the gingival crevicular fluid (GCF) and saliva (whole stimulated saliva) of orthodontic patients at different time points after fixed appliance activation. Methods: MPO was determined in the GCF and collected by means of periopaper from the saliva of 14 patients with orthodontic fixed appliances. GCF and saliva samples were collected at baseline, 2 hours, and 7 and 14 days after application of the orthodontic force. Results: Mean MPO activity was increased in both the GCF and saliva of orthodontic patients at 2 hours after appliance activation (P<0.02 for all comparisons). At 2 hours, PMN infiltration into the periodontal ligament from the orthodontic force probably results in the increased MPO level observed at this time point. Conclusions: MPO might be a good marker to assess inflammation in orthodontic movement; it deserves further studies in orthodontic therapy. (Am J Orthod Dentofacial Orthop 2010;138:613-6)

Saliva flow rate, buffer capacity, and pH of autistic individuals

The objective of the study was to evaluate saliva flow rate, buffer capacity, pH levels, and dental caries experience (DCE) in autistic individuals, comparing the results with a control group (CG). The study was performed on 25 noninstitutionalized autistic boys, divided in two groups. G1 composed of ten children, ages 3-8. G2 composed of 15 adolescents ages 9-13. The CG was composed of 25 healthy boys, randomly selected and also divided in two groups: CG3 composed of 14 children ages 4-8, and CG4 composed of 11 adolescents ages 9-14. Whole saliva was collected under slight suction, and pH and buffer capacity were determined using a digital pHmeter. Buffer capacity was measured by titration using 0.01 N HCl, and the flow rate expressed in ml/min, and the DCE was expressed by decayed, missing, and filled teeth (permanent dentition [DMFT] and primary dentition [dmft]). Data were plotted and submitted to nonparametric (Kruskal-Wallis) and parametric (Student`s t test) statistical tests with a significance level less than 0.05. When comparing G1 and CG3, groups did not differ in flow rate, pH levels, buffer capacity, or DMFT. Groups G2 and CG4 differ significantly in pH (p = 0.007) and pHi = 7.0 (p = 0.001), with lower scores for G2. In autistic individuals aged 3-8 and 9-13, medicated or not, there was no significant statistical difference in flow rate, pH, and buffer capacity. The comparison of DCE among autistic children and CG children with deciduous (dmft) and mixed/permanent decayed, missing, and filled teeth (DMFT) did not show statistical difference (p = 0.743). Data suggest that autistic individuals have neither a higher flow rate nor a better buffer capacity. Similar DCE was observed in both groups studied.

Effects of purified herbal extract of Saliva miltiorrhiza on ischemic rat myocardium after acute myocardial infarction

In the current study, we compared purified Salvia miltiorrhiza extract (PSME) with Angiotensin-converting enzyme inhibitor, Ramipril, in in vitro experiments and also in vivo using animal model of myocardial infarction. PSME was found to have a significantly higher trolox equivalent antioxidant capacity which indicated a great capacity for scavenging free radicals. PSME could also prevent pyrogallo red bleaching and DNA damage.

After 2 weeks treatment with PSME or Ramipril, survival rates of rats with experimental myocardial infarction were marginally increased (68.2% and 71.4%) compared with saline (61.5%). The ratios of infarct size to left ventricular size in both PSME-and Ramipril-treated rats were significantly less than that in the saline-treated group. Activity of cardiac antioxidant enzyme superoxide dismutase (SOD) was significant higher while level of Thiobarbituric acid-reactive substances (TBARs) was lower in the PSME treated group. Purified and standardized Chinese herb could provide an alternative regimen for the prevention of ischemic heart disease.

Stability of cp-Ti and Ti-6Al-4V alloy for dental implants as a function of saliva pH - an electrochemical study

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparação entre a utilização de saliva e sangue para determinação do lactato mínimo em cicloergômetro e ergômetro de braço em mesa-tenistas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Correlation among mutans streptococci counts, dental caries, and IgA to Streptococcus mutans in saliva

Two-hundred and forty individuals were studied, divided into five groups as follows: caries-free children, children with caries, children with rampant caries, young adults with and without caries. Whole stimulated saliva was collected and all individuals were investigated for DMFT/dmft according to the WHO criteria and the simplified oral hygiene index (OHI-S). Quantitative analysis of the total aerobic flora and mutans streptococci in saliva was performed. Also, the level of salivary anti-S. mutans IgA was determined by ELISA. Children with rampant caries showed the highest OHI-S value. The highest total counts of microorganisms were found in the group of children with caries. No statistically significant differences were observed for salivary flow, OHI-S and microorganism counts between the groups of young adults. No correlation between mutans streptococci counts and anti-Streptococcus mutans IgA levels was observed in the studied groups. A correlation between increased anti-Streptococcus mutans IgA levels and caries-free status was observed among young adults but not among children.

Adherence of Streptococcus mutans to uncoated and saliva-coated glass-ceramics and composites

This study sought to investigate the surface roughness and the adherence of Streptococcus mutans (in the presence and absence of saliva) to ceramics and composites. The early dental biofilms formed in situ on the materials were illustrated, using scanning electron microscopy (SEM). Feldspathic and leucite/feldspathic ceramics and microhybrid and microfilled composites were evaluated. Human dental enamel was used as the control. Standardized specimens of the materials were produced and surface roughness was analyzed. The adhesion tests were carried out in 24-well plates and colony forming units (CFU/mL) were evaluated. Values of roughness (μm) and adherence (CFU/mL) were analyzed statistically. Of all the surfaces tested, enamel was the roughest. Leucite/feldspathic ceramics were rougher than the feldspathic ceramic, while composites were similar statistically. Enamel offered the highest level of adherence to uncoated and saliva-coated specimens, while the leucite/feldspathic ceramic demonstrated greater adherence than the feldspathic ceramic and the composites were similar statically. The rougher restorative materials increased the adherence of S, mutans on the material surfaces.

Avaliação das correlações existentes entre experiência de cárie, acúmulo de placa bacteriana e nível de Iga anti-streptococcus mutans na saliva de crianças com dentição mista

Pós-graduação em Odontologia Restauradora - ICT

Aumento da atividade dos sistemas antioxidantes modula o estresse oxidativo na saliva de crianças com cárie precoce severa

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Functional imaging of the parotid glands using blood oxygenation level dependent (BOLD)-MRI at 1.5T and 3T

PURPOSE: To evaluate the function of the parotid glands before and during gustatory stimulation, using an intrinsic susceptibility-weighted MRI method (blood oxygenation level dependent, BOLD-MRI) at 1.5T and 3T. MATERIALS AND METHODS: A total of 10 and 13 volunteers were investigated at 1.5T and 3T, respectively. Measurements were performed before and during gustatory stimulation using ascorbate. Circular regions of interest (ROIs) were delineated in the left and right parotid glands, and in the masseter muscle for comparison. The effects of stimulation were evaluated by calculating the difference between the relaxation rates, DeltaR(2)*. Baseline and stimulation were statistically compared (Student's t-tests), merging both parotid glands. RESULTS: The averaged DeltaR(2)* values prestimulation obtained in all parotid glands were stable (-0.61 to 0.38 x 10(-3) seconds(-1)). At 3T, these values were characterized by an initial drop (to -2.7 x 10(-3) seconds(-1)) followed by a progressive increase toward the baseline. No significant difference was observed between baseline and parotid gland stimulation at 1.5T, neither for the masseter muscle at both field strengths. A considerable interindividual variability (over 76%) was noticed at both magnetic fields. CONCLUSION: BOLD-MRI at 3T was able to detect DeltaR(2)* changes in the parotid glands during gustatory stimulation, consistent with an increase in oxygen consumption during saliva production.