Proteins In Saliva Research Articles

Integrative analysis of transcriptome and proteome in primary Sjögren syndrome

ObjectivePrimary Sjögren's syndrome (pSS) is a intricate autoimmune disease mainly characterized of immune-mediated destruction of exocrine tissues, such as salivary and lacrimal glands, occurring dry mouth and eyes. Although some breakthroughs in understanding pSS have been uncovered, many questions remain about its pathogenesis, especially the internal relations between exocrine glands and secretions. MethodTranscriptomic and proteomic analyses were conducted on salivary tissues and saliva in experimental Sjögren syndrome (ESS). The ESS model was established by immunization with salivary gland protein. The expression of mRNAs and proteins in salivary tissues and saliva were determined by high-throughput sequencing transcriptomic analysis and LC-MS/MS-based proteome, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were used to recognize dysregulated genes and proteins. The association between RNA and protein abundance was investigated to provides a comprehensive understanding of RNA-protein correlations in the pathogenesis of pSS. ResultsAs a result, we successfully established the ESS model. We recognized 3221 differentially expressed genes (DEGs) and 253 differentially expressed proteins (DEPs). The sample analysis showed that 61 proteins overlapped through the integrative analysis of transcriptomics and proteomics data. The enrichment pathway analysis of DEGs and DEPs in samples showed alterations in renin-angiotensin-system (RAS), lysosome, and apoptosis. Notably, we found that some genes, such as AGT, FN1, Klk1b26, Klk1, Klk1b5, Klk1b3 had a consistent trend in the regulation at the RNA and protein levels and might be potential diagnostic biomarkers of pSS. ConclusionHerein, we found critical processes and potential biomakers that may contribute to pSS pathogenesis by analyzing dysregulated genes and pathways. Additionally, the integrative multi-omics datasets provided additional insight into understanding complicated disease mechanisms.

LC‒MS/MS and transcriptome analyses reveal saliva components of the seed-feeding truebug Pyrrhocoris apterus

Saliva secretion is essential for successful feeding by piercing–sucking insects. Despite extensive studies of the saliva composition of phloem- and blood-feeding insects, less is known about the oral secretions of seed-feeding insects. The firebug Pyrrhocoris apterus is a polyphagous insect that feeds primarily on dry seeds of various plant species. Here, liquid chromatography‒mass spectrometry/mass spectrometry (LC‒MS/MS) was used to identify 165 P. apterus salivary proteins, including 110 gelling-specific proteins, 24 watery-specific proteins, and 31 proteins common to both groups. Transcriptomic analysis of saliva glands identified 567 potential salivary proteins, of which 87 overlapped with those detected by LC‒MS/MS. Comparative analysis showed that 98 out of 165 (59.4%) P. apterus saliva proteins were associated with extra-oral digestion and detoxification and had homologs in ten other species of arthropods examined. In addition, 67 proteins were specific to P. apterus, of which 22 proteins were defined with unknown functions. RNA interference-mediated gene silencing assays indicated that P. apterus-specific proteins were vital for P. apterus survival. Our findings shed light on the function of salivary proteins in feeding by seed-feeding insects and the evolution of feeding habits in piercing–sucking insects.

A Point-of-Care Testing Device Utilizing Graphene-Enhanced Fiber Optic SPR Sensor for Real-Time Detection of Infectious Pathogens.

Timely detection of highly infectious pathogens is essential for preventing and controlling public health risks. However, most traditional testing instruments require multiple tedious steps and ultimately testing in hospitals and third-party laboratories. The sample transfer process significantly prolongs the time to obtain test results. To tackle this aspect, a portable fiber optic surface plasmon resonance (FO-SPR) device was developed for the real-time detection of infectious pathogens. The portable device innovatively integrated a compact FO-SPR sensing component, a signal acquisition and processing system, and an embedded power supply unit. A gold-plated fiber is used as the FO-SPR sensing probe. Compared with traditional SPR sensing systems, the device is smaller size, lighter weight, and higher convenience. To enhance the detection capacity of pathogens, a monolayer graphene was coated on the sensing region of the FO-SPR sensing probe. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was used to evaluate the performance of the portable device. The device can accurately detect the SARS-CoV-2 spike S1 protein in phosphate-buffered saline (PBS) and artificial saliva within just 20 min, and the device successfully detected cultured SARS-CoV-2 virus. Furthermore, the FO-SPR probe has long-term stability, remaining stable for up to 8 days. It could distinguish between the SARS-CoV-2 spike protein and the MERS-CoV spike protein. Hence, this FO-SPR device provides reliable, rapid, and portable access to test results. It provides a promising point-of-care testing (POCT) tool for on-site screening of infectious pathogens.

Salivary microbiome changes distinguish response to chemoradiotherapy in patients with oral cancer

BackgroundOral squamous cell carcinoma (SCC) is associated with oral microbial dysbiosis. In this unique study, we compared pre- to post-treatment salivary microbiome in patients with SCC by 16S rRNA gene sequencing and examined how microbiome changes correlated with the expression of an anti-microbial protein.ResultsTreatment of SCC was associated with a reduction in overall bacterial richness and diversity. There were significant changes in the microbial community structure, including a decrease in the abundance of Porphyromonaceae and Prevotellaceae and an increase in Lactobacillaceae. There were also significant changes in the microbial community structure before and after treatment with chemoradiotherapy, but not with surgery alone. In patients treated with chemoradiotherapy alone, several bacterial populations were differentially abundant between responders and non-responders before and after therapy. Microbiome changes were associated with a change in the expression of DMBT1, an anti-microbial protein in human saliva. Additionally, we found that salivary DMBT1, which increases after treatment, could serve as a post-treatment salivary biomarker that links to microbial changes. Specifically, post-treatment increases in human salivary DMBT1 correlated with increased abundance of Gemella spp., Pasteurellaceae spp., Lactobacillus spp., and Oribacterium spp. This is the first longitudinal study to investigate treatment-associated changes (chemoradiotherapy and surgery) in the oral microbiome in patients with SCC along with changes in expression of an anti-microbial protein in saliva.ConclusionsThe composition of the oral microbiota may predict treatment responses; salivary DMBT1 may have a role in modulating the oral microbiome in patients with SCC.Graphical After completion of treatment, 6 months after diagnosis, patients had a less diverse and less rich oral microbiome. Leptotrichia was a highly prevalent bacteria genus associated with disease. Expression of DMBT1 was higher after treatment and associated with microbiome changes, the most prominent genus being Gemella-6tcCpfJtMWsEpEDBUWdC3Video

Smartphone-assisted paper-based electrochemical immunosensor for SARS-CoV-2 detection in saliva

Herein, we developed a new waste solution-free paper-based electrochemical immunosensor for SARS-CoV-2 detection in saliva, by combining vertical and lateral flow. In detail, the device was constituted of a reservoir containing all reagents for the construction of the immunological chain onto the magnetic beads and a lateral flow holder which contained a polyester-based electrode, a magnet, and an adsorbent pad. The measurement was carried out by adding the saliva sample into the reservoir, followed by the addition of this solution in the hole present in the lateral flow holder. The successive additions of washing buffer and TMB solution in the lateral flow holder allowed the detection of N protein in saliva in the range of 0.06 to 4 µg/mL with a detection limit equal to 30 ng/mL. The analysis of several saliva samples with the sensing tool and the reference method, demonstrated the effectiveness of this device, being able to identify positive patients with high values of CT e.g. 35. This new configuration paves the way for the realization of any magnetic beads-based immunosystem without waste solution production, enlarging the application of paper-based devices.

Salivary IgA and vimentin differentiate in vitro SARS-CoV-2 infection: A study of 290 convalescent COVID-19 patients

SARS-CoV-2 initially infects cells in the nasopharynx and oral cavity. The immune system at these mucosal sites plays a crucial role in minimizing viral transmission and infection. To develop new strategies for preventing SARS-CoV-2 infection, this study aimed to identify proteins that protect against viral infection in saliva. We collected 551 saliva samples from 290 healthcare workers who had tested positive for COVID-19, before vaccination, between June and December 2020. The samples were categorized based on their ability to block or enhance infection using in vitro assays. Mass spectrometry and enzyme-linked immunosorbent assay experiments were used to identify and measure the abundance of proteins that specifically bind to SARS-CoV-2 antigens. Immunoglobulin (Ig)A specific to SARS-CoV-2 antigens was detectable in over 83% of the convalescent saliva samples. We found that concentrations of anti-receptor-binding domain IgA >500 pg/µg total protein in saliva correlate with reduced viral infectivity in vitro. However, there is a dissociation between the salivary IgA response to SARS-CoV-2, and systemic IgG titers in convalescent COVID-19 patients. Then, using an innovative technique known as spike-baited mass spectrometry, we identified novel spike-binding proteins in saliva, most notably vimentin, which correlated with increased viral infectivity in vitro and could serve as a therapeutic target against COVID-19.

Could mosquito bites facilitate IgE sensitization to food and inhalant allergens?

We present an hypothesis that arthropod bites and stings can induce sensitization to concomitantly introduced food and inhalant allergens in human, and that contents of the sting and bite such as venom and saliva proteins act as adjuvants further facilitating sensitization. In food allergen sensitization, evidence presently supports the hypothesis that exposure through a disrupted skin-barrier is allergenic, and exposure to food proteins through ingestion is tolerogenic. Stinging and biting arthropods that breach skin barrier are known to provide IgE sensitization to their own venom and salivary allergens e. g. Hymenoptera and mosquito. Murine experiments have shown that content of venom from honeybee (phospholipase 2 (PLA2) and hyaluronidase) facilitate IgE sensitization to a concomitantly introduced food allergen in the skin. Conserved biomolecules in saliva and venom present in several arthropod species (e.g. PLAs and hyaluronidases) are suggested to play a role as adjuvants in sensitization to concomitantly presented allergens during arthropod stings and bites. Tick bites have been shown to provide IgE sensitization to tick salivary allergens and IgE sensitization to non-primate mammalian red blood cell allergen Galactose-alpha-1,3-galactose, causing allergy to non-primate red meat, termed Alpha-gal syndrome (AGS).Mosquitos may take up allergens such as pathogenesis related proteins (e.g. lipid transfer proteins (LTPs), and PR-10) when feeding on plant nectar and sap and may take up animal derived allergens such as immunoglobulins, serum albumins and lipocalins from feeding on blood. It is proposed that plant and animal allergens, important in human allergic disease are carried on mouthparts or in gut of mosquitos and transferred into the skin of a human host during a mosquito bite. We hypothesize that sensitization to common food and inhalant allergens can occur facilitated by a mosquito bite, possibly facilitated by adjuvant factors such as PLA2 and hyaluronidase in mosquito saliva. The hypothesis may provide insight into basic mechanisms in IgE sensitization.

Metaproteogenomic analysis of saliva samples from Parkinson’s disease patients with cognitive impairment

Cognitive impairment (CI) is very common in patients with Parkinson’s Disease (PD) and progressively develops on a spectrum from mild cognitive impairment (PD-MCI) to full dementia (PDD). Identification of PD patients at risk of developing cognitive decline, therefore, is unmet need in the clinic to manage the disease. Previous studies reported that oral microbiota of PD patients was altered even at early stages and poor oral hygiene is associated with dementia. However, data from single modalities are often unable to explain complex chronic diseases in the brain and cannot reliably predict the risk of disease progression. Here, we performed integrative metaproteogenomic characterization of salivary microbiota and tested the hypothesis that biological molecules of saliva and saliva microbiota dynamically shift in association with the progression of cognitive decline and harbor discriminatory key signatures across the spectrum of CI in PD. We recruited a cohort of 115 participants in a multi-center study and employed multi-omics factor analysis (MOFA) to integrate amplicon sequencing and metaproteomic analysis to identify signature taxa and proteins in saliva. Our baseline analyses revealed contrasting interplay between the genus Neisseria and Lactobacillus and Ligilactobacillus genera across the spectrum of CI. The group specific signature profiles enabled us to identify bacterial genera and protein groups associated with CI stages in PD. Our study describes compositional dynamics of saliva across the spectrum of CI in PD and paves the way for developing non-invasive biomarker strategies to predict the risk of CI progression in PD.

Multiplexed detection of viral antigen and RNA using nanopore sensing and encoded molecular probes

We report on single-molecule nanopore sensing combined with position-encoded DNA molecular probes, with chemistry tuned to simultaneously identify various antigen proteins and multiple RNA gene fragments of SARS-CoV-2 with high sensitivity and selectivity. We show that this sensing strategy can directly detect spike (S) and nucleocapsid (N) proteins in unprocessed human saliva. Moreover, our approach enables the identification of RNA fragments from patient samples using nasal/throat swabs, enabling the identification of critical mutations such as D614G, G446S, or Y144del among viral variants. In particular, it can detect and discriminate between SARS-CoV-2 lineages of wild-type B.1.1.7 (Alpha), B.1.617.2 (Delta), and B.1.1.539 (Omicron) within a single measurement without the need for nucleic acid sequencing. The sensing strategy of the molecular probes is easily adaptable to other viral targets and diseases and can be expanded depending on the application required.

Metabolomic changes associated with acquired resistance to Ixodes scapularis

Guinea pigs repeatedly exposed to Ixodes scapularis develop acquired resistance to the ticks (ATR). The molecular mechanisms of ATR have not been fully elucidated, and partially involves immune responses to proteins in tick saliva. In this study, we examined the metabolome of sera of guinea pigs during the development of ATR. Induction of components of the tyrosine metabolic pathway, including hydroxyphenyllactic acid (HPLA), were associated with ATR. We therefore administered HPLA to mice, an animal that does not develop ATR, and exposed the animals to I. scapularis. We also administered nitisinone, a known inhibitor of tyrosine degradation, to another group of mice. The mortality of I. scapularis that fed on mice given HPLA or nitisinone was 26 % and 72 % respectively, compared with 2 % mortality among ticks that fed on control animals. These data indicate that tick bites alter the guinea pig metabolome, and that the tyrosine metabolism pathway can potentially be targeted for I. scapularis control.

Difference in astringency of the main pea protein fractions

Interactions between food and saliva govern complex mouthfeel perceptions such as astringency. Herein, we present a study of the interactions of salivary proteins with the main pea protein fractions that are obtained by isoelectric and salt precipitation (legumin-rich, vicilin-rich and albumin-rich fractions). The sensory evaluations performed on protein solutions by trained panelists evidenced that all three protein fractions exhibit a basal level of astringency, but that the albumin fraction was perceived as the most astringent one. All three fractions induced significant but comparable loss of salivary lubrication. Yet, when compared to the other fractions, the albumin fraction showed the formation of a thicker and more rigid film on salivary conditioning film-coated sensors as measured using a quartz crystal microbalance with dissipation monitoring (QCM-D). We also present proteomics studies on the precipitates obtained from the mixtures of saliva and pea protein fractions. Protein identification finds a pool of salivary proteins involved in non-specific interactions with all the three pea protein fractions. Yet, 13 pea proteins specific to the albumin fraction were identified as being involved in specific interactions with salivary proteins. Several of these proteins are part of the plant defense mechanisms and are likely to interact with many salivary proteins. This could explain the higher number of salivary proteins found in the precipitate induced by the albumin fraction when compared to the other two. These quantitative results increase the understanding of the complex links between plant protein-salivary protein interactions and astringency.

Remineralizing gel with calcium lactate for the prevention of caries in children

The article presents the results of a single-center open clinical trial of the use of the remineralizing gel «PRESIDENT Minerals gel 0+». The effectiveness of the drug in children in the period of mixed dentition with a compensated form of caries was studied.Materials and methods. The study involved 19 children, mean age 8,3 years. The duration of the study is 1 month.Clinical indices and laboratory parameters were determined: DMF+df, OHI-S, PMA, the degree of enamel electrometry, calcium, glucose, total protein and antioxidant activity of saliva (AOA). Organoleptic properties were determined on a 5-point scale.Results. After one month of using the remineralizing gel, a decrease in the hygiene index by 23,27%, a decrease in the electrometric evaluation of enamel by 15,58% were revealed. In saliva, a decrease in the level of protein by 33,13% and an increase in the content of calcium by 53,33% were revealed. The organoleptic properties of the gel were evaluated on a five-point scale at 4,31±0,22 points.Conclusion. «PRESIDENT 0+ Minerals gel» with calcium lactate and hydroxyapatite has a remineralizing effect, good organoleptic properties and can be recommended for caries prevention in children.

Ixodes scapularis nymph saliva protein blocks host inflammation and complement-mediated killing of Lyme disease agent, Borrelia burgdorferi.

Tick serine protease inhibitors (serpins) play crucial roles in tick feeding and pathogen transmission. We demonstrate that Ixodes scapularis (Ixs) nymph tick saliva serpin (S) 41 (IxsS41), secreted by Borrelia burgdorferi (Bb)-infected ticks at high abundance, is involved in regulating tick evasion of host innate immunity and promoting host colonization by Bb. Recombinant (r) proteins were expressed in Pichia pastoris, and substrate hydrolysis assays were used to determine. Ex vivo (complement and hemostasis function related) and in vivo (paw edema and effect on Bb colonization of C3H/HeN mice organs) assays were conducted to validate function. We demonstrate that rIxsS41 inhibits chymase and cathepsin G, pro-inflammatory proteases that are released by mast cells and neutrophils, the first immune cells at the tick feeding site. Importantly, stoichiometry of inhibition analysis revealed that 2.2 and 2.8 molecules of rIxsS41 are needed to 100% inhibit 1 molecule of chymase and cathepsin G, respectively, suggesting that findings here are likely events at the tick feeding site. Furthermore, chymase-mediated paw edema, induced by the mast cell degranulator, compound 48/80 (C48/80), was blocked by rIxsS41. Likewise, rIxsS41 reduced membrane attack complex (MAC) deposition via the alternative and lectin complement activation pathways and dose-dependently protected Bb from complement killing. Additionally, co-inoculating C3H/HeN mice with Bb together with rIxsS41 or with a mixture (rIxsS41 and C48/80). Findings in this study suggest that IxsS41 markedly contributes to tick feeding and host colonization by Bb. Therefore, we conclude that IxsS41 is a potential candidate for an anti-tick vaccine to prevent transmission of the Lyme disease agent.

The utility of salivary CRP and IL-6 as a non-invasive measurement evaluated in patients with COVID-19 with and without diabetes

Background: The available evidence suggests that inflammatory responses, in both systemic and oral tissue, contribute to the pathology of COVID-19 disease. Hence, studies of inflammation biomarkers in oral fluids, such as saliva, might be useful to better specify COVID-19 features. Methods: In the current study, we performed quantitative real-time PCR to measure salivary levels of C-reactive protein (CRP) and interleukin-6 (IL-6) in saliva obtained from patients diagnosed with mild COVID-19, in a diabetic group (DG; n = 10) and a non-diabetic group (NDG; n = 13). All participants were diagnosed with periodontitis, while six participants with periodontitis but not diagnosed with COVID-19 were included as controls. Results: We found increases in salivary total protein levels in both the DG and NDG compared to control patients. In both groups, salivary CRP and IL-6 levels were comparable. Additionally, the levels of salivary CRP were significantly correlated with total proteins, in which a strong and moderate positive correlation was found between DG and NDG, respectively. A linear positive correlation was also noted in the relationship between salivary IL-6 level and total proteins, but the correlation was not significant. Interestingly, the association between salivary CRP and IL-6 levels was positive. However, a moderately significant correlation was only found in COVID-19 patients with diabetes, through which the association was validated by a receiver operating curve. Conclusions: These finding suggest that salivary CRP and IL-6 are particularly relevant as potential non-invasive biomarker for predicting diabetes risk in mild cases of COVID-19 accompanied with periodontitis.

Evaluation of CRP, IL-6 and Calprotectin in Saliva of Patients Suffering From Crohn’s Disease (CD)

Inflammatory bowel disease (IBD) a chronic inflammatory condition of the gastrointestinal tract characterized by alternating episodes of clinical relapse and remission (Wallace, 2014), with Crohn's disease (CD) and ulcerative colitis (UC) being the two primary subtypes (Souza, 2016). The present study was achieved in Baghdad teaching hospital from November 2021 to June 2022 to prove if the diagnostic marker of Crohn’s disease (CD) (IL-6, CRP and Calprotectin( are rises in saliva as in sera and if there are significant difference in the levels of those markers between the Crohn’s disease (CD)group and control group.
 The recent study was measured the levels of C-reactive protein (CRP), Interleukin-6 (IL6) and calprotectin in saliva of CD patients using ELISA method as those proteins has been previously proved to be elevated in sera of CD patient.
 The results of the present study noted a significant mean differences in salivary levels of each of CRP,IL6 and calprotectin between CD group and the control group ( P < 0.05 ).
 The current study was proved that there is a significant positive correlation between IL-6 levels and age in all study groups (CD and control) (P <0.05) and also a significant positive correlation was found between IL-6 and CRP in CD group patients (p<0.05) respectively.

Blue-emitting SiO2-coated Si-doped ZnSeS quantum dots conjugated aptamer-molecular beacon as an electrochemical and metal-enhanced fluorescence biosensor for SARS-CoV-2 spike protein

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) which was first reported in early January 2020, continues to devastate the worlds public health system. Herein, we report on the development of a novel metal-enhanced fluorescence (MEF) and electrochemical biosensor for SARS-CoV-2 spike (S) protein. To develop the MEF biosensor, SiO2-coated Si-doped ZnSeS quantum dots (QDs) were newly synthesized and conjugated to an aptamer-molecular beacon (Apta-MB) probe. Thereafter, cationic AuNPs, used as a localised surface plasmon resonance (LSPR) signal amplifier, were self-assembled on the QDs-Apta-MB conjugate to form a QDs-Apta-MB-AuNP probe. To develop the electrochemical biosensor, the QDs-Apta-MB assay was carried out on a carbon nanofiber-modified screen-printed carbon electrode. Cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS) were used to characterize the electrode surface whilst spectrophotometric, spectroscopic, fluorescence polarization and electron microscopic techniques were used to characterize the materials. Under optimal experimental conditions, the QDs binding to the Apta-MB, quenched the QDs’ fluorescence and with SARS-CoV-2 S protein binding to the Apta-MB, LSPR signal from cationic AuNPs of different sizes and shapes were used to tune the fluorescence signal to obtain enhanced sensitivity. On the other hand, using [Fe(CN)6]/K3−/4- buffered with NaAc-KAc-TrizmaAc-KSCN-Borax as the electrolyte solution, anodic peaks of the QDs from the CV and DPV plots were unravelled. Electrochemical detection of SARS-CoV-2 S protein was accomplished by a systematic increase in the QDs anodic peak current generated from the DPV plots. The limits of detection obtained for the SARS-CoV-2 S protein were 8.9 fg/mL for the QDs-Apta-MB-AuNP MEF probe and ∼0.5 pg/mL for the QDs-Apta-MB electrochemical probe. Detection of SARS-CoV-2 S protein in saliva was demonstrated using the QDs-Apta-MB-AuNP MEF probe.

An Analytical Tooth Model Based on Surface Plasmon Resonance Chips Coated with Hydroxyapatite Used for Investigation of the Acquired Dental Pellicle

One of the main targets in modern dentistry is the prevention of caries. Not only daily nutrition plays an important role, but also the so‐called “pellicle” on the outer surface of teeth, which is mainly formed by saliva proteins. Moreover, numerous bacteria are part of this pellicle, which might cause caries by acidic metabolites. In the here‐presented study, a method for electrophoretic deposition (EPD) of hydroxyapatite (HAP) nanoparticles on gold surfaces, suitable for surface plasmon resonance (SPR) measurements, is developed. An “artificial tooth” is created and loaded by natural saliva in different concentrations (to form a natural pellicle), while the influence of an environment with different pH values has been studied. It could be demonstrated that even slight acid solutions damage the pellicle significantly within seconds, exposing the HAP to acidic degradation, which would lead to caries in the human oral cavities. This model allows to study pellicle formation as well as degradation in real time. As a practical example, the influence of beverages on the pellicle could be demonstrated.

Proteomic analysis of stimulated saliva in gastroesophageal reflux disease patients with and without erosive tooth wear: Observational study

ObjectiveTo evaluate the difference in the proteomic profile of stimulated saliva in patients with gastroesophageal reflux disease (GERD) with (GE) and without (GNE) erosive tooth wear (ETW), regarding both human and bacterial proteins. MethodsStimulated saliva (SS) was collected from 16 patients (8/group). Samples were centrifuged at 4.500 g for 15 min under refrigeration to remove all debris. The supernatant from each saliva sample was taken and frozen at -80 °C. After extracting the proteins, they were submitted to reverse phase liquid chromatography and mass spectrometry (nLC-ESI-MS/MS). Label-free proteomic quantification was performed using Protein Lynx Global Service (PLGS) software (p < 0.05) for human and bacterial proteins. ResultsIn total, 67 human proteins were common for GNE and GE groups. GNE group presented, compared to GE group, increase in proteins that confer antimicrobial and acid resistant properties, such as cystatins, histatin and immunoglobulins. However, GNE group had a marked decrease in subunits of hemoglobin (α, β and delta). Regarding bacterial proteins, for SS, 7 and 10 unique proteins were identified in the GE and GNE groups, respectively. They are related to protein synthesis and energy metabolism and interact with human proteins typically found in saliva and supramolecular complexes of the acquired pellicle. ConclusionsOur data indicate that the stimulation of the salivary flow increases acid resistant and antimicrobial proteins in saliva, which might protect against ETW. Clinical significanceThis pioneer study showed important differences in the human and bacterial proteome of SS in patients with GERD with or without ETW.

Arabinogalactan proteins and polysaccharides compete directly with condensed tannins for saliva proteins influencing astringency perception of Cabernet Sauvignon wines

Wine astringency is thought to be due to salivary protein precipitation; however, the actual mechanism is not well-defined. This study aimed understand the relationship between whole polysaccharide extracts, produced with and without enzyme maceration, and the saliva protein-tannin precipitation reaction. Polysaccharides were analyzed in the context of salivary protein–tannin interactions using gel electrophoresis, quantitative 1H proton nuclear magnetic resonance (qHNMR), size separation chromatography, immunochemistry, and sensory analysis. Polysaccharide addition reduced saliva protein concentration in tannin-saliva protein-polysaccharide mixtures, indicating that native-wine polysaccharides compete with condensed tannins for salivary protein as ligand partners. qHNMR showed that tannin levels were increased by adding polysaccharides, suggesting that in these conditions, polysaccharides interact with saliva proteins via competitive protein-polysaccharide complex formation. Polysaccharides from non-enzyme-treated wines had threshold concentration of 121 mg/mL versus 86 mg/ml for enzyme-treated as detected by a sensory panel. Enzyme-treated polysaccharides changed astringency perception at a lower concentration than non-enzyme-treated polysaccharides.

Variation of wine preference amongst consumers is influenced by the composition of salivary proteins

The preferences of consumers for different flavours and aromas in wine are varied and may be explained by inherent factors such as cultural background, wine education and personal taste of the wine consumer. Wine flavour, as perceived in the mouth, includes aroma compounds released through the retronasal pathway, which are shaped by interactions with saliva. Saliva and wine interactions could provide an explanation as to why wine tasters express different preferences for wine. To test this hypothesis, 13 Western and 13 Chinese experienced wine tasters were recruited. Sensory evaluation was performed in formal surroundings to acquire free description-based and perceived sensory intensity data using the Pivot® Profile and continuous scale assessment, respectively. Participants’ saliva samples were collected before the sensory evaluation and spiked into a wine sample to investigate the impact on the wine’s volatile release using comprehensive two-dimensional gas chromatography–mass spectrometry (GC × GC–MS). Saliva samples were subjected to enzyme activity assays and protein composition profiling by Tandem Mass Tag (TMT) quantitative proteomics. The wine tasters showed differences in wine flavour perception, which was supported by the difference in wine volatile release resulting from the addition of saliva. The two groups of participants did not have significant differences in total salivary protein concentrations or the amounts of esterase and α-amylase. However, statistically significant variations in the concentrations of specific proteins (proline-rich proteins (PRPs) and lipocalin-1 (LCN-1); p < 0.01) were found between the two groups. Significant correlations between perceived intensities of wine attributes and concentrations of PRPs and LCN-1 were observed. These results indicate that the composition of proteins in saliva is a factor that influences wine perception and preference. Our results provide a biochemical basis for understanding preference for food based on interactions between aroma compounds and salivary proteins and could be used to suggest foods or beverages to particular cultural groups.