Analysis Of Saliva Research Articles

Sample Preparation for Metabolomic Analysis in Exercise Physiology

Metabolomics investigates final and intermediate metabolic products in cells. Assessment of the human metabolome relies principally on the analysis of blood, urine, saliva, sweat, and feces. Tissue biopsy is employed less frequently. Understanding the metabolite composition of biosamples from athletes can significantly improve our knowledge of molecular processes associated with the efficiency of training and recovery. Such knowledge may also lead to new management opportunities. Successful execution of metabolomic studies requires simultaneous qualitative and quantitative analyses of numerous small biomolecules in samples under test. Unlike genomics and proteomics, which do not allow for direct assessment of enzymatic activity, metabolomics focuses on biochemical phenotypes, providing unique information about health and physiological features. Crucial factors in ensuring the efficacy of metabolomic analysis are the meticulous selection and pre-treatment of samples.

Rapid and non-invasive analysis of paracetamol overdose using paper arrow-mass spectrometry: a prospective observational study

BackgroundParacetamol is the most consumed medicine globally. Its accessibility contributes to common overdose. Paracetamol overdose is responsible for > 50% of acute liver failure cases, making it the second most common reason for a liver transplant. Rapid quantitation of paracetamol is crucial to guide treatment of paracetamol overdose. Current tests require invasive sampling and relatively long turnaround times. Paper arrow-mass spectrometry (PA-MS) combines sample collection, extraction, separation, enrichment and ionisation onto a single paper strip, achieving rapid, accurate, cost-effective and eco-friendly analysis direct from raw human saliva.MethodsTo validate PA-MS against an established test, 17 healthy adults were recruited. Samples were collected before and at 15, 30, 60, 120 and 240 min after ingesting 1 g of paracetamol. Plasma measured with an established clinical test served as the reference standard to validate PA-MS with three biofluids—plasma, resting saliva (RS) and stimulated saliva (SS). Participants’ views of blood, RS and SS sampling procedures were assessed qualitatively. Cross-validation was assessed using Lin’s concordance correlation coefficients (CCC), Bland–Altman difference plots, and ratios of PA-MS to the reference standard test.ResultsPA-MS using stimulated saliva offers a reliable alternative to intravenous blood sampling. The CCC is 0.93, the mean difference with the reference test is − 0.14 mg/L, and the ratios compared to the reference test are 0.84–1.27 from correlated samples collected at 5 intervals over 4 h for each participant.ConclusionsParacetamol detection from SS with PA-MS provides a reliable result that can aid timely treatment decisions. Differences between paracetamol concentration in resting and stimulated saliva were also identified for the first time, highlighting the importance of standardising saliva collection methods in general. This study marks a major milestone towards rapid and convenient saliva analysis.

Insights into Medication-Induced Osteonecrosis of the Jaw Through the Application of Salivary Proteomics and Bioinformatics.

Long-term treatment with bisphosphonates is accompanied by an increased risk of medication-related osteonecrosis of the jaw (MRONJ). Currently, no clinically useful biomarkers for the predictive diagnosis of MRONJ are available. To investigate the potential key proteins involved in the pathogenesis of MRONJ, a proteomic LC-MS/MS analysis of saliva was performed. Differentially expressed proteins (DEPs) were analyzed using BiNGO, ClueGO, cytoHubba, MCODE, KEGG, and ReactomeFI software packages using Cytoscape platforms. In total, 1545 DEPs were identified, including 43 up- and 11 down-regulated with a 1.5-fold cut-off value and adj. p-value < 0.05. The analysis provided a panel of hub genes, including APOA2, APOB, APOC2, APOC3, APOE, APOM, C4B, C4BPA, C9, FGG, GC, HP, HRG, LPA, SAA2-SAA4, and SERPIND1. The most prevalent terms in GO of the biological process were macromolecular complex remodeling, protein-lipid complex remodeling, and plasma lipoprotein particle remodeling. DEPs were mainly involved in signaling pathways associated with lipoproteins, the innate immune system, complement, and coagulation cascades. The current investigation advanced our knowledge of the molecular mechanisms underlying MRONJ. In particular, the research identified the principal salivary proteins that are implicated in the onset and progression of this condition.

The effect of caffeine dose on caffeine and paraxanthine changes in serum and saliva and CYP1A2 enzyme activity in athletes: a randomized placebo-controlled crossover trial

BackgroundAlthough caffeine (CAF) supplementation has been shown to improve exercise performance, its dose-dependent effect on CAF metabolism has not been sufficiently investigated. The aim of this study was to evaluate the effects of 3, 6 and 9 mg of CAF/kgBM on changes of CAF and paraxanthine (PRX) in the serum and saliva at four time-points.MethodsIn a randomized, double-blind, placebo-controlled crossover design, acute pre-exercise supplementation in 26 moderately-trained athletes, participating in high-intensity functional training (HIFT), was examined. The study protocol involved CAF/PRX biochemical analyses of serum and saliva with respect to CYP1A2 polymorphism and CYP1A2 enzyme activity.ResultsDespite significant differences between the serum and saliva levels of CAF and PRX, there was no difference in the PRX/CAF ratio. The interaction effect of dose and time-points for PRX concentration was revealed. The main effects of dose were observed for CAF and the PRX/CAF ratio. The main effect of time-points was registered only for serum CAF.ConclusionsDose- and time-dependent effect of CAF supplementation on CAF and PRX in the serum and saliva of athletes was confirmed, but there was no effect of the CAF dose on CYP1A2 enzyme activity, nor was there an interaction of CYP1A2 with enzyme inducibility. The CAF/PRX correlation indicated the possibility of interchangeable use of serum and/or saliva analyses in exercise studies.Clinical trial registrationThis trial was registered prospectively at ClinicalTrials.gov (NCT03822663, registration date: 30/01/2019).

Expression of Salivary miRNAs, Clinical, and Demographic Features in the Early Detection of Gastric Cancer: A Statistical and Machine Learning Analysis.

Gastric cancer ranks as one of the top five deadliest cancers worldwide and is often diagnosed at late stages. Analysis of saliva may provide a non-invasive approach for detection of malignancies in organs associated with the oral cavity. This research aims to analyze salivary microRNA expression together with clinical and demographic features with the aim of diagnosing gastric cancer. The study included 19 patients with early-stage gastric cancer and 19 healthy controls. Saliva samples were collected and processed for RNA isolation. Salivary expression of miR-223-3p and miR-21-5p were measured using quantitative reverse-transcription polymerase chain reaction (RT-qPCR). Receiver operating characteristic (ROC) curves were generated to evaluate the accuracy of diagnostic models. Machine learning algorithms, multiple logistic regression, and principal component analysis (PCA) were used to assess the predictive power of miRNAs in conjunction with clinical-demographic features. Significant upregulation of miR-223-3p and downregulation of miR-21-5p in saliva were observed in patients with gastric cancer. The area under ROC curve (AUC) values for salivary miR-21-5p, salivary miR-223-3p, and their multiple logistic regression were determined to be 0.723, 0.791, and 0.850, respectively. The AUC for multiple logistic regression model was 0.919. The PCA model led to the highest diagnostic odds ratio (DOR) of 134.33 (sensitivity = 0.785, specificity = 1.00, AUC = 903). Application of machine learning methods, and in particular a random forest algorithm, showed high accuracy in diagnosing patients with gastric cancer (sensitivity = 1.00, specificity = 0.857, AUC = 0.93). The application of validated salivary diagnostics in clinical practice could help facilitate earlier diagnosis of gastric cancer and improve medical outcome. Expression of miR-21 and miR-223-3p in saliva together with clinical and demographic features, appears promising in screening for GC.

On the effectiveness of virtual reality technology for relaxation of locomotive crew workers

Reducing the risks of occupational stress requires improving the methods of psychological support for employees of locomotive crews. Virtual reality technology offers restorative measures comparable to traditional techniques. The study aims to analyze the effectiveness of the impact of virtual reality technology on the basis of the analysis of the psycho-emotional state of employees of locomotive crews before and after rehabilitation measures; to evaluate the ratio of stress biomarkers before and after exposure to classical and technical techniques; to compare the effectiveness of psychological influence carried out by various methods. The scientists conducted a survey and diagnosis of the functional state of locomotive crew workers, followed by rehabilitation measures using various methods. The specialists performed a saliva analysis based on the concentration levels of cortisol and dehydroepiandrosterone. 120 people took part in the study, of which: 60 people according to the classical program and 60 people according to the virtual reality program, of which 15 people are at risk according to psychophysiological indicators. The session lasted 21 minutes. The average age of employees of locomotive crews participating in the study was 35.4 years. 88% of the subjects underwent functional state diagnostics and a full course of relaxation sessions in virtual reality. Of these, 73% have improved psychophysiological indicators: response accuracy, increased neuropsychiatric stability, and decreased anxiety levels. The authors noted calmness, improved mood and relief of muscle tension. 60% of the employees have completed a full course of rehabilitation measures using classical techniques. Of these, 70% have an improvement in their functional state according to certain parameters. There is a decrease in cortisol levels (unit of measurement — nanograms per milliliter) from 6.75 ng/ml to 6.28 ng/ml with a simultaneous increase in dehydroepiadrosterone levels (unit of measurement — micrograms per milliliter) from 0.53 mcg/100 ml to 1.77 mcg/100 ml, which corresponds to a decrease in the overall stress level. The analysis of the obtained results showed sufficient effectiveness of virtual methods comparable to classical approaches to relaxation. Immersive technologies more effectively activate natural compensatory mechanisms by expanding the individual regulatory capabilities of the nervous system. Scientists have identified areas for improving the latest methods of rehabilitation measures in order to correct the functional condition of locomotive crews. Limitations. The representativeness of the sample was ensured by the participation of 120 employees of locomotive crews in the study. The limitations are related to the gender homogeneity of the sample: only men are included, due to their professional status. Ethics. The study was performed in accordance with the rules of good clinical practice and the Helsinki Declaration of the World Medical Association, the conclusion of the ethics committee was not required.

Prospective evaluation of non-invasive saliva specimens for the diagnosis of syphilis and molecular surveillance of Treponema pallidum.

The promising diagnostic performance of molecular testing for syphilis using saliva and urine samples has been reported; however, further evaluation of its possible application for diagnosis and molecular surveillance is required. In addition, the development of a rapid and easy-to-perform molecular test for syphilis is important for its use in the clinical setting. We comprehensively evaluated the diagnostic and surveillance performance of two novel loop-mediated isothermal amplification (LAMP) assays using saliva and urine samples. Saliva, urine, and whole blood were collected from patients who underwent serological testing for syphilis at outpatient clinics. Treponema pallidum DNA in specimens was detected using quantitative PCR (qPCR), nested PCR, and novel LAMP assays. T. pallidum genotyping was conducted by multi-locus sequence typing (MLST). Of the 163 patients recruited, 98 were diagnosed with syphilis (primary: n = 35; secondary: n = 40; latent: n = 23). qPCR showed the highest sensitivity among the molecular tests performed with a sensitivity of 54.1% and 30.3% for all syphilis patients using saliva and urine samples, respectively. A novel method of LAMP combined with dry reagents and crude DNA extraction (Dry-LAMP) showed a probit detection limit of 37.4 copies/reaction within 45 min. The agreement rate between Dry-LAMP and qPCR for saliva was 95.7% (κ coefficient 0.90). The T. pallidum genotype was identified in 48 patients by MLST using saliva samples. Molecular analysis of saliva could be used as a supplementary diagnostic test for syphilis and molecular surveillance of the T. pallidum genotype. Dry-LAMP is expected to be helpful in the clinical diagnosis of syphilis.

The Impact of Triclosan-Containing Toothpaste on the Overall and Cariogenic Salivary Microbiota Among Primary School Children

Objectives: Triclosan-diphenyl ether derivate is an agent, which belongs to the Non-Ionic antimicrobial agents group. This study aims to estimate the salivary levels of Streptococcus mutans and Lactobacillus species in the saliva before and after teeth brushing with a dentifrice containing triclosan/copolymer and to compare the number of whole salivary flora by saliva analysis before and after teeth brushing. Methods: To accomplish our objective, we used a Colgate Total Plus Whitening®, Colgate-Palmolive dentifrice. The group consisted of 24 healthy schoolchildren aged 9 – 13 of both sexes. The counts of Streptococcus mutans and Lactobacillus species were determined with commercial CRT bacteria strips produced by Ivoclar-Vivadent, Liechtenstein. The total count of the saliva microbial was determined by standard microbiological methods. Results: A significant reduction in salivary MS and LB levels was observed in all samples as well as a decrease in the total count of aerobe and anaerobe bacteria and yeast. Conclusion: The effectiveness of Colgate Total Plus Whitening® toothpaste in reducing salivary aerobes is statistically significant, but its impact on reducing salivary anaerobic microbes is statistically negligible. The bacteria Streptococcus mutans experiences a decrease that is greater than tenfold, while the bacterium Lactobacillus species experiences a reduction of around tenfold.

Anti-cariogenic effect of experimental resin cement containing ursolic acid using dental microcosm biofilm

ObjectiveThis study aimed to assess the anticariogenic effects of resin cement containing varying ursolic acid (UA) concentrations and to determine the optimal UA concentrations in the microcosm biofilm model. Materials and methodsExperimental resin cements with UA concentrations of 0, 0.1, 0.5, 1.0, and 2.0 wt% were prepared. Class I cavities were prepared on 50 extracted human molars and restored with composite inlays and experimental resin cements. Tooth samples were subjected to artificial caries induction for 10 days in a microcosm biofilm model using human saliva as an inoculum, and then mineral changes were evaluated using quantitative light-induced fluorescence (ΔF and ΔQ) and micro-computed tomography (CT). The bacterial composition of the human saliva was analyzed by 16 s RNA microbiome profiling. One-way analysis of variance with Tukey and Duncan post-hoc tests was employed for statistical analysis (p < 0.05). ResultsAs the UA concentration increased, resin cement decreased ΔF and ΔQ before and after caries induction but showed a significant difference only in ΔQ at UA concentration ≥ 1.0 % (p < 0.05). The gray value analysis result of micro CT also showed a significant difference at UA concentration ≥ 1.0 % (p < 0.05). In the human saliva analysis, bacterial composition remained within normal oral microbiota ranges. ConclusionResin cements containing at least 1.0 % of UA exhibited an anticariogenic effect on dental microcosm biofilms. Clinical relevanceTo reduce the failure of restorations, it is essential to prevent the occurrence of secondary caries. The application of UA in resin cement can be utilized to prevent the formation of secondary caries due to the anticariogenic effect of UA.

Salivary Metabolic Pathway Alterations in Brazilian E-Cigarette Users.

In recent years, the use of electronic cigarettes (e-cigs) has increased. However, their long-term effects on oral health and saliva remain poorly understood. Therefore, this study aimed to evaluate the saliva of e-cig users and investigate possible biomarkers. Participants were divided into two groups: the Electronic Cigarette Group (EG)-25 regular and exclusive e-cig users-and Control Group (CG)-25 non-smokers and non-e-cig users, matched in sex and age to the EG. The clinical analysis included the following parameters: age, sex, heart rate, oximetry, capillary blood glucose, carbon monoxide (CO) concentration in exhaled air, and alcohol use disorder identification test (AUDIT). Qualitative and quantitative analyses of saliva included sialometry, viscosity, pH, and cotinine concentrations. Furthermore, the EG and CG salivary metabolomes were compared using gas chromatography coupled with mass spectrometry (GC-MS). Data were analyzed using the Mann-Whitney test. The MetaboAnalyst 6.0 software was used for statistical analysis and biomarker evaluation. The EG showed high means for exhaled CO concentration and AUDIT but lower means for oximetry and salivary viscosity. Furthermore, 10 metabolites (isoleucine, 2-hydroxyglutaric acid, 3-phenyl-lactic acid, linoleic acid, 3-hydroxybutyric acid, 1,6-anhydroglucose, glucuronic acid, valine, stearic acid, and elaidic acid) were abundant in EG but absent in CG. It was concluded that e-cig users had high rates of alcohol consumption and experienced significant impacts on their general health, including increased cotinine and CO concentration in exhaled air, decreased oximetry, and low salivary viscosity. Furthermore, they showed a notable increase in salivary metabolites, especially those related to inflammation, xenobiotic metabolism, and biomass-burning pathways.

Elucidation of pathophysiology of intracranial aneurysms based on metagenome analyses of oral bacteria

When intracranial aneurysms occur in arteries, this can lead to weakening of the arterial wall and subarachnoid haemorrhage, which can result in death. Intracranial aneurysms can’t be diagnosed without cerebrovascular imaging tests and there are few clues to indicate their presence. Neurosurgeon Assistant Professor Hirokazu Koseki, Department of Neurosurgery, The Jikei University School of Medicine, Japan, is looking for novel ways to predict the presence of intracranial aneurysms before they rupture. He is investigating the role of oral bacteria, which have been associated with cerebral haemorrhage and dementia, in order to clarify the relationship between oral bacteria and aneurysms. In unprecedented work, he and the team want to map this relationship based on severity of periodontal disease and will utilise metagenomic analysis of saliva to identify the bacterial strains that might contribute to the neuropathology. They will also take samples from the walls of the aneurysms to precisely decipher which bacterial strains can be directly associated with them. The work will enable researchers to establish simple and inexpensive targeted treatment for intracranial aneurysms and is also anticipated to have knock-on effects, being used for the screening of inherent risk factors for the occurrence of intracranial aneurysms. Although there are known risk factors, patients without any of the risk factors can still develop intracranial aneurysms or subarachnoid haemorrhages. Building on existing research that points to association between oral microbiology and neuropathology, Koseki is working to identify further risk factors that might help in diagnosing intracranial aneurysms in patients.

Utility of Raman Spectroscopy in Pulmonary Medicine.

The Raman effect, or as per its original description, "modified scattering", is an observation that the number of scattered light waves shifts after photons make nonelastic contact with a molecule. This effect allows Raman spectroscopy to be very useful in various fields. Although it is well known that Raman spectroscopy could be very beneficial in medicine as a diagnostic tool, there are not many applications of Raman spectroscopy in pulmonary medicine. Mostly tumor tissue, sputum and saliva have been used as material for analysis in respiratory medicine. Raman spectroscopy has shown promising results in malignancy recognition and even tumor staging. Saliva is a biological fluid that could be used as a reliable biomarker of the physiological state of the human body, and is easily acquired. Saliva analysis using Raman spectroscopy has the potential to be a relatively inexpensive and quick tool that could be used for diagnostic, screening and phenotyping purposes. Chronic obstructive pulmonary disease (COPD) is a growing cause of disability and death, and its phenotyping using saliva analysis via Raman spectroscopy has a great potential to be a dependable tool to, among other things, help reduce hospitalizations and disease burden. Although existing methods are effective and generally available, Raman spectroscopy has the benefit of being quick and noninvasive, potentially reducing healthcare costs and workload.

Fungiform Papillae and Gustatory Function in Neurofibromatosis Type 1: A Case-Control Study.

Fungiform papillae enlargement is a common oral manifestation of neurofibromatosis type 1 (NF1). This study aimed to objectively evaluate the size, number, and symmetry of fungiform papillae in NF1 individuals and investigate the relationship between these alterations and taste perception, salivary flow, dietary habits, and BMI. A cross-sectional case-control study was conducted on 80 participants (40 with NF1 and 40 controls), matched by age and sex. Participants underwent quantitative and morphological evaluation of fungiform papillae, gustatory perception tests, sialometry, saliva analysis, xerostomia assessment, dietary assessments, and Body Mass Index calculations. The NF1 group exhibited significantly larger and more asymmetric fungiform papillae and exhibited a higher detection threshold for sweet and sour tastes, as well as hyposalivation and lower preference for healthy foods compared to the controls. No correlation was found between papillae morphology, gustatory perception tests, saliva properties, xerostomia, food preferences, or BMI in the NF1 group. Enlarged and asymmetric fungiform papillae, hyposalivation, heightened sensitivity to sweet and sour tastes, and reduced healthy eating habits were common in NF1. Although fungiform papillae alterations seem unrelated to taste sensitivity and food preferences, further investigation is needed to better understand the mechanisms underlying these changes.

Saliva as a diagnostic tool in soccer: a scoping review.

A high-performance sport like soccer requires training strategies that aim to reach peak performance at the right time for the desired competitions. Thus, the investigation of biochemical markers in saliva is a tool that is beginning to be used in athletes within the physical training process. There is still no evidence on universal saliva collection and analysis protocols in soccer. This review aims to map the use of saliva as a tool for analyzing athletic performance in soccer, from the biomarkers used to the validated protocols for these analyses. A broad systematic literature search was carried out in the electronic databases Web of Science, Livivo, Scopus, PubMed, LILACS and gray literature (Google Scholar and ProQuest). Two reviewers selected the studies and extracted data on the type of salivary collection used, the salivary biomarker evaluated and monitored. Ninety-three articles were included. The most frequently analyzed salivary biomarkers were cortisol (n=53), testosterone (n=35), secretory immunoglobulin A (SIgA) (n=33), salivary alpha amylase (n=7), genetic polymorphisms (n=4) and miRNAs (n=2). The results of the studies indicated beneficial effects in monitoring salivary biomarkers in the assessment of sports performance, although most studies did not include a control group capable of comparison. Salivary collection and analysis protocols were varied and commonly not reported. This scoping review provides a comprehensive overview of the current landscape of salivary biomarker research in soccer. The findings underscore the importance of these biomarkers in assessing athletes' physiological responses and overall well-being. Future research should focus on refining methodologies, exploring additional biomarkers, and investigating the practical implications of salivary biomarker monitoring in soccer and other sports.

An enzymatic reaction-based SERS saliva analysis microporous array chip for chiral differentiation and high-throughput detection of D-amino acids.

ARaman-active boronate modified surface-enhanced Raman scattering (SERS) microporous array chip based on the enzymatic reaction was constructed for reliable, sensitive, and quantitative monitoring of D-Proline (D-Pro) and D-Alanine (D-Ala) in saliva. Initially, 3-mercaptophenylboronic acid (3-MPBA) was bonded to Au-coated Si nanocrown arrays (Au/SiNCA) via Au-S bonding. Following this, H2O2 obtained from D-amino acid oxidase (DAAO)-specific catalyzed D-amino acids (D-AAs) further reduced 3-MPBA to 3-hydroxythiophenol (3-HTP) with a new Raman peak at 882cm-1. Meanwhile, the original characteristic peak at 998cm-1 remained unchanged. Therefore, the I882/I998 ratio increased with increasing content ofD-AAs in the sample to be tested, allowing D-AAs to be quantitatively detected. TheAu/SiNCA with large-area periodic crown structure prepared provided numerous, uniform "hot spots," and the microporous array chip with 16 detection units was employed as the platform for SERS analysis, realizing high-throughput, high sensitivity, high specificity and high-reliability quantitative detection of D-AAs (D-Pro and D-Ala). The limits of detection (LOD) were down to 10.1µM and 13.7µM throughout the linear range of 20-500µM. The good results of the saliva detection suggested that this SERS sensor could rapidly differentiate between early-stage gastric cancer patients and healthy individuals.

12260 Assessing The Impact Of Saliva Collection Devices On Analyte Recovery

Abstract Disclosure: D. Marshall: None. M. Dolan: None. M. Mctaggart: None. B.G. Keevil: None. Introduction: Analysis of saliva is gathering momentum, often conveying advantages over regular venepuncture, offering a convenient way of collecting multiple samples throughout the day with relative ease from the comfort of a patient’s home or workplace. The advent of more sensitive mass spectrometry equipment has opened the possibility of measuring analytes previously unsuitable, offering a multitude of steroid hormones for diagnosis and monitoring of a range of endocrine disorders. Traditionally, samples are collected onto a Sarstedt Salivette device comprising of a synthetic swab enclosed within a polypropylene tube. For certain analytes, the Salivette device is known to impede analyte recovery, resulting in the requirement for a passive drool sample which can be difficult and unpleasant to collect for some patients. Methods Previous work has been undertaken showing the SalivaBio Oral Swab (SOS) to have less of an impact on analyte recovery. Here, through addition of low, medium and high spikes and subsequent LC-MS/MS analysis of 19 different steroid hormones (11-dehydrocorticosterone, 11-deoxycorticosterone, 11-deoxycortisol, 11-hydroxyandrostenedione, 11-hydroxytestosterone, 11-ketoandrostenedione, 11-ketotestosterone, 17-hydroxyprogesterone, 18-hydroxycortisol, 18-oxocortisol, 21-deoxycortisol, aldosterone, androstenedione, corticosterone, cortisol, cortisone, dexamethasone, testosterone and tetrahydro11-deoxycortisol) we aimed to assess a broader repertoire of analytes and whether saliva collection devices impact on analyte recovery. Results With an acceptability criterion of 10% difference from baseline passive drool samples, Salivette devices were acceptable for 12 out of 18 analytes, SOS were acceptable for 16 analytes. Both devices were unacceptable for measurement of androstenedione, testosterone and 11-deoxycorticosterone. When 10% is deemed as an acceptable difference, the SOS performs better than the Salivette device for the steroid hormones investigated in this study. This may have implications for research studies, patient diagnosis and monitoring of endocrine disorders where saliva collection devices were previously deemed unacceptable due to issues with analyte recovery. Presentation: 6/2/2024

Attenuated Total Reflection-Fourier Transform Infrared (ATR-FTIR) Spectroscopy Analysis of Saliva as a Diagnostic Specimen for Rapid Classification of Oral Squamous Cell Carcinoma Using Chemometrics Methods

Background & Aim Recent advancements in analytical techniques have highlighted the potential of Attenuated Total Reflection-Fourier Transform Infrared (ATR-FTIR) spectroscopy as a quick, cost-effective, non-invasive, and efficient tool for cancer diagnosis. This study aims to evaluate the effectiveness of ATR-FTIR spectroscopy in combination with supervised machine learning classification models for diagnosing OSCC using saliva samples. Methods & Materials Eighty unstimulated whole saliva samples from OSCC patients and healthy controls were collected. The ATR-FTIR spectroscopy was performed and spectral data were used to classify healthy and OSCC groups. The data were analyzed using machine learning classification methods such as Partial Least Squares-Discriminant Analysis (PLS-DA) and Support Vector Machine Classification (SVM-C). The classification performance of the models was evaluated by computing sensitivity, specificity, precision, and accuracy. Results The samples were classified into two classes based on their spectral data. The obtained results demonstrate a high level of accuracy in the prediction sets of the PLS-DA and SVM-C models, with accuracy values of 0.960 and 0.962, respectively. The OSCC group sensitivity values for both PLS-DA and SVM-C models was 1.00, respectively. Conclusion The study indicates that ATR-FTIR spectroscopy, combined with chemometrics, is a potential method for the non-invasive diagnosis of OSCC using saliva samples. This method achieved high accuracy and the findings of this study suggest that ATR-FTIR spectroscopy could be further developed for clinical applications in OSCC diagnosis.

Oral squamous cell carcinoma identification by FTIR spectroscopy of oral biofluids.

This case study evaluated the efficacy of mid-infrared spectroscopy on the identification of oral squamous cell carcinoma, following the assessment of unstimulated whole saliva. The trial follows a matched case-control design. Saliva samples were characterized through mid-infrared spectroscopy, and chemometric tools were applied to distinguish between case and control participants, further identifying the spectral regions that played a pivotal role in the successful identification of oral squamous cell carcinoma. Mid-infrared spectroscopy was capable to discriminate between cancer patients and matched controls with 100% of correct predictions. Additionally, the spectral regions mostly contributing to the successful prediction were identified and found to be potentially associated with significant molecular changes crucial to the carcinogenic process. The application of mid-infrared spectroscopy in saliva analysis may be regarded as an innovative, noninvasive, low cost, and sensitive technique contributing to the identification of oral squamous cell carcionma.

Electrochemical immunoplatform for the determination of multiple salivary biomarkers of oral diseases related to microbiome dysbiosis

Several diseases of the oral cavity are related to compositional and functional shifts in the oral microbiome. The analysis of saliva is an attractive alternative for the diagnosis and prognosis of these diseases. Samples can be obtained by no invasive procedures and processing is relatively simple. However, sensitive and selective analytical methods are needed to make the diagnosis as specific as possible. In this work, four salivary biomarkers of oral diseases: interleukin-6 (IL-6), receptor activator of NF-kB ligand (RANKL), protein arginine deiminase 4 (PAD4) and the corresponding antibody (anti-PAD-4) were selected as targets for their simultaneous determination using an electrochemical immunosensing platform. Sandwich-type amperometric immunoassays were implemented using horseradish peroxidase (HRP)/H2O2/hydroquinone (HQ) for application to the analysis of saliva of six volunteers. The developed method provides excellent sensitivity, selectivity, and wide linear ranges with LOD values of 0.09 pg mL−1 (IL-6), 0.10 pg mL−1 (RANKL); 0.09 ng mL−1(PAD4) and 14.5 ng mL−1 (anti-PAD4) and allows the accurate analysis of saliva without matrix effects, using 25 μL of raw sample. The developed methodology is competitive with commercial ELISA kits available only for a single biomarker determination, while the assay for the four biomarkers can be completed in less than two hours.

A PhD completed. Oral manifestations in inflammatory bowel diseases

Crohn's disease and ulcerative colitis are chronic inflammatory diseases of the gastrointestinal tract. In addition to bowel symptoms, patients may also have oral manifestations. This thesis investigated potential associations between disease activity in the gut, oral health, salivary gland function, and saliva composition. Patients with Crohn's disease had a significantly higher DMFT index, but showed no difference in periodontal diseases compared to a healthy control group. The saliva composition in patients with active bowel disease differed from that in patients with inactive bowel disease, suggesting that saliva analysis could potentially be used in the future to determine the degree and severity of bowel disease. The knowledge of gastroenterologists and dentists regarding oral manifestations of bowel diseases was found to be limited. Gastroenterologists and dentists valued interdisciplinary patient consultation as very useful, but the frequency of consultation was considered insufficient.