Salivary Metabolomics:From Diagnostic Biomarker Discovery to Investigating Biological Function

Metabolomic profiling of biofluids, e.g., urine, plasma, has generated vast and ever-increasing amounts of knowledge over the last few decades. Paradoxically, metabolomic analysis of saliva, the most readily-available human biofluid, has lagged. This review explores the history of saliva-based metabolomics and summarizes current knowledge of salivary metabolomics. Current applications of salivary metabolomics have largely focused on diagnostic biomarker discovery and the diagnostic value of the current literature base is explored. There is also a small, albeit promising, literature base concerning the use of salivary metabolomics in monitoring athletic performance. Functional roles of salivary metabolites remain largely unexplored. Areas of emerging knowledge include the role of oral host–microbiome interactions in shaping the salivary metabolite profile and the potential roles of salivary metabolites in oral physiology, e.g., in taste perception. Discussion of future research directions describes the need to begin acquiring a greater knowledge of the function of salivary metabolites, a current research direction in the field of the gut metabolome. The role of saliva as an easily obtainable, information-rich fluid that could complement other gastrointestinal fluids in the exploration of the gut metabolome is emphasized

Developing and Standardizing a Protocol for Quantitative Proton Nuclear Magnetic Resonance ( ¹ H NMR) Spectroscopy of Saliva

Metabolic profiling by ¹H NMR spectroscopy is an underutilized technology in salivary research, although preliminary studies have identified promising results in multiple fields (diagnostics, nutrition, sports physiology). Translation of preliminary findings into validated, clinically approved knowledge is hindered by variability in protocol for the collection, storage, preparation, and analysis of saliva. This study aims to evaluate the effects of differing sample pretreatments on the ¹H NMR metabolic profile of saliva. Protocol considerations are highly varied in the current literature base, including centrifugation, freeze–thaw cycles, and different NMR quantification methods. Our findings suggest that the ¹H NMR metabolite profile of saliva is resilient to any change resulting from freezing, including freezing of saliva prior to centrifuging. However, centrifugation was necessary to remove an unidentified broad peak between 1.24 and 1.3 ppm, the intensity of which correlated strongly with saliva cellular content. This peak obscured the methyl peak from lactate and significantly affected quantification. Metabolite quantification was similar for saliva centrifuged between 750<i>g</i> to 15 000<i>g</i>. Quantification of salivary metabolites was similar whether quantified using internal phosphate-buffered sodium trimethylsilyl-[2,2,3,3-²H₄]-propionate (TSP) or external TSP in a coaxial NMR tube placed inside the NMR tube containing the saliva sample. Our results suggest that the existing literature on salivary ¹H NMR will not have been adversely affected by variations of the common protocol; however, use of TSP as an internal standard without a buffered medium appears to affect metabolite quantification, notably for acetate and methanol. We include protocol recommendations to facilitate future NMR-based studies of saliva

Endogenous salivary citrate is associated with enhanced rheological properties following oral capsaicin-stimulation

NEW FINDINGS: What is the central question of this study? What are the relationships between physical properties of saliva, protein composition and metabolite composition? What is the main finding and its importance? Salivary citrate, one of the major endogenous metabolites in saliva, increased upon capsaicin stimulation and was associated with improved physical properties measured by extensional rheology. This suggests salivary gland citrate transporters might be a valuable area of future study.ABSTRACT: Saliva displays viscoelastic properties which enable coating, lubrication and protection of the oral mucosa and hard tissues. Individuals lacking saliva or perceiving oral dryness can manage their symptoms using artificial saliva preparations, but these often fail to mimic the sensation and functionality of natural saliva. It is widely acknowledged that mucins (MUC7 and MUC5B) confer saliva's rheological properties, but artificial saliva containing purified mucins is still often an inadequate substitute. This work aimed to explore salivary components that influence salivary extensional rheology to better understand how natural saliva could be replicated. Saliva was stimulated via control and capsaicin solutions in healthy volunteers. Extensional rheology was analysed using a CaBER-1 (capillary breakup) extensional rheometer. Protein composition, including mucins, was measured by gel-electrophoresis band densitometry and metabolites were measured by 1 H nuclear magnetic resonance spectroscopy. Capsaicin stimulation significantly increased capillary breakup time, extensional viscosity and the abundance of most major salivary proteins. Stimulation also increased salivary citrate and choline concentrations. Significant correlations were found between capillary breakup time and amylase (r = 0.67, P &lt; 0.05), statherin (ρ = 0.66, P &lt; 0.05) and citrate (ρ = 0.81, P &lt; 0.01). The relationship between citrate and salivary rheology was subsequently investigated in vitro. These results suggest that citrate and non-mucin proteins are stronger predictors of salivary rheology than the more often studied mucin glycoproteins. Potential mechanisms are discussed and future work in this area could help formulate more effective saliva substitutes, more closely resembling natural saliva.</p

Determining bacterial and host contributions to the human salivary metabolome

BACKGROUND: Salivary metabolomics is rapidly advancing. AIM AND METHODS: To determine the extent to which salivary metabolites reflects host or microbial metabolic activity whole-mouth saliva (WMS), parotid saliva (PS) and plasma collected contemporaneously from healthy volunteers were analysed by (1)H-NMR spectroscopy. Spectra underwent principal component analysis and k-means cluster analysis and metabolite quantification. WMS samples were cultured on both sucrose and peptide-enriched media. Correlation between metabolite concentration and bacterial load was assessed. RESULTS: WMS contained abundant short-chain fatty acids (SCFAs), which were minimal in PS and plasma. WMS spectral exhibited greater inter-individual variation than those of PS or plasma (6.7 and 3.6 fold, respectively), likely reflecting diversity of microbial metabolomes. WMS bacterial load correlated strongly with SCFA levels. Additional WMS metabolites including amines, amino acids and organic acids were positively correlated with bacterial load. Lactate, urea and citrate appeared to enter WMS via PS and the circulation. Urea correlated inversely with WMS bacterial load. CONCLUSIONS: Oral microbiota contribute significantly to the WMS metabolome. Several WMS metabolites (lactate, urea and citrate) are derived from the host circulation. WMS may be particularly useful to aid diagnosis of conditions reflective of dysbiosis. WMS could also complement other gastrointestinal fluids in future metabolomic studies

GAME-UP: Game-Aware Mode Enumeration and Understanding for Trajectory Prediction

Interactions between road agents present a significant challenge in trajectory prediction, especially in cases involving multiple agents. Because existing diversity-aware predictors do not account for the interactive nature of multi-agent predictions, they may miss these important interaction outcomes. In this paper, we propose GAME-UP, a framework for trajectory prediction that leverages game-theoretic inverse reinforcement learning to improve coverage of multi-modal predictions. We use a training-time game-theoretic numerical analysis as an auxiliary loss resulting in improved coverage and accuracy without presuming a taxonomy of actions for the agents. We demonstrate our approach on the interactive subset of Waymo Open Motion Dataset, including three subsets involving scenarios with high interaction complexity. Experiment results show that our predictor produces accurate predictions while covering twice as many possible interactions versus a baseline model.Comment: 10 pages, 6 figure

No Association between HIV and Intimate Partner Violence among Women in 10 Developing Countries

Intimate Partner Violence (IPV) has been reported to be a determinant of women's risk for HIV. We examined the relationship between women's self-reported experiences of IPV in their most recent relationship and their laboratory-confirmed HIV serostatus in ten low- to middle-income countries.Data for the study came from the most recent Demographic and Health Surveys conducted in Dominican Republic, Haiti, India, Kenya, Liberia, Malawi, Mali, Rwanda, Zambia and Zimbabwe. Each survey population was a cross-sectional sample of women aged 15-49 years. Information on IPV was obtained by a face-to-face interview with the mother with an 81.1% response rate; information on HIV serostatus was obtained from blood samples with an 85.3% response rate. Demographic and socioeconomic variables were considered as potentially confounding covariates. Logistic regression models accounting for multi-stage survey design were estimated individually for each country and as a pooled total with country fixed effects (n = 60,114). Country-specific adjusted odds ratios (OR) for physical or sexual IPV compared to neither ranged from 0.45 [95% confidence interval (CI): 0.23-0.90] in Haiti to 1.35 [95% CI: 0.95-1.90] in India; the pooled association was 1.03 [95% CI: 0.94-1.13]. Country-specific adjusted ORs for physical and sexual IPV compared to no sexual IPV ranged from 0.41 [95% CI: 0.12-1.36] in Haiti to 1.41 [95% CI: 0.26-7.77] in Mali; the pooled association was 1.05 [95% CI: 0.90-1.22].IPV and HIV were not found to be consistently associated amongst ever-married women in national population samples in these lower income countries, suggesting that IPV is not consistently associated with HIV prevalence worldwide. More research is needed to understand the circumstances in which IPV and HIV are and are not associated with one another

Measurement and Interpretation of Fermion-Pair Production at LEP energies above the Z Resonance

This paper presents DELPHI measurements and interpretations of cross-sections, forward-backward asymmetries, and angular distributions, for the e+e- -> ffbar process for centre-of-mass energies above the Z resonance, from sqrt(s) ~ 130 - 207 GeV at the LEP collider. The measurements are consistent with the predictions of the Standard Model and are used to study a variety of models including the S-Matrix ansatz for e+e- -> ffbar scattering and several models which include physics beyond the Standard Model: the exchange of Z' bosons, contact interactions between fermions, the exchange of gravitons in large extra dimensions and the exchange of sneutrino in R-parity violating supersymmetry.Comment: 79 pages, 16 figures, Accepted by Eur. Phys. J.

A Determination of the Centre-of-Mass Energy at LEP2 using Radiative 2-fermion Events

Using e+e- -> mu+mu-(gamma) and e+e- -> qqbar(gamma) events radiative to the Z pole, DELPHI has determined the centre-of-mass energy, sqrt{s}, using energy and momentum constraint methods. The results are expressed as deviations from the nominal LEP centre-of-mass energy, measured using other techniques. The results are found to be compatible with the LEP Energy Working Group estimates for a combination of the 1997 to 2000 data sets.Comment: 20 pages, 6 figures, Accepted by Eur. Phys. J.

A Measurement of the Tau Hadronic Branching Ratios

The exclusive and semi-exclusive branching ratios of the tau lepton hadronic decay modes (h- v_t, h- pi0 v_t, h- pi0 pi0 v_t, h- \geq 2pi0 v_t, h- \geq 3pi0 v_t, 2h- h+ v_t, 2h- h+ pi0 v_t, 2h- h+ \geq 2pi0 v_t, 3h- 2h+ v_t and 3h- 2h+ \geq 1pi0 v_t) were measured with data from the DELPHI detector at LEP.Comment: 53 pages, 18 figures, Accepted by Eur. Phys. J.