Biomarkers In Saliva Research Articles

Combining PDMS Composite and Plasmonic Solid Chemosensors: Dual Determination of Ammonium and Hydrogen Sulfide as Biomarkers in a Saliva Single Test

In recent years, in the field of bioanalysis, the use of saliva as a biological fluid for the determination of biomarkers has been proposed. Saliva analysis stands out for its simplicity and non-invasive sampling. This paper proposes a method for the dual determination of ammonium and hydrogen sulfur in saliva using two colorimetric chemosensors. The ammonia reacts with 1,2-Naftoquinone 4 sulphonic acid (NQS) entrapped in polydimethylsiloxane (PDMS) and the hydrogen sulfide with AgNPs retained in a nylon membrane. The color changed from orange to brown in the case of ammonia chemosensors and from yellow to brown in the H2S. The experimental conditions to be tested have been established. Both analytes have been determined from their gaseous form; these are ammonia from ammonium and hydrogen sulfur from hydrogen sulfur. Good figures of merit have been obtained by using both measuring strategies (reflectance diffuse and digitalized images). The acquired results show that both sensors can be used and provide good selectivity and sensitivity for the determination of these biomarkers in saliva. Both measurement strategies have provided satisfactory results for the real saliva samples (n = 15). Recoveries on spiked samples were between 70 and 100%. This methodology can lead to possible in situ diagnosis and monitoring of certain diseases and pathologies related with NH4+ and/or H2S, in a fast, simple, cheap and non-invasive way.

Non-invasive early detection on esophageal squamous cell carcinoma and precancerous lesions by microbial biomarkers combining epidemiological factors in China.

Microbiota may be associated with esophageal squamous cell carcinoma (ESCC) development. However, it is not known the predictive value of microbial biomarkers combining epidemiological factors for the early detection of ESCC and precancerous lesions. A total of 449 specimens (esophageal swabs and saliva) were collected from 349 participants with different esophageal statuses in China to explore and validate ESCC-associated microbial biomarkers from genes level to species level by 16S rRNA sequencing, metagenomic sequencing and real-time quantitative polymerase chain reaction. A bacterial biomarker panel including Actinomyces graevenitzii (A.g_1, A.g_2, A.g_3, A.g_4), Fusobacteria nucleatum (F.n_1, F.n_2, F.n_3), Haemophilus haemolyticus (H.h_1), Porphyromonas gingivalis (P.g_1, P.g_2, P.g_3) and Streptococcus australis (S.a_1) was explored by metagenomic sequencing to early detect the participants in Need group (low-grade intraepithelial neoplasia, high-grade intraepithelial neoplasia and ESCC) vs participants without these lesions as the Noneed group. Significant quantitative differences existed for each microbial target in which the detection efficiency rate was higher in saliva than esophageal swab. In saliva, the area under the curve (AUC) based on the microbial biomarkers (A.g_4 ∩ P.g_3 ∩ H.h_1 ∩ S.a_1 ∩ F.n_2) was 0.722 (95% CI 0.621-0.823) in the exploration cohort. Combining epidemiological factors (age, smoking, drinking, intake of high-temperature food and toothache), the AUC improved to 0.869 (95% CI 0.802-0.937) in the exploration cohort, which was validated with AUC of 0.757 (95% CI 0.663-0.852) in the validation cohort. It is feasible to combine microbial biomarkers in saliva and epidemiological factors to early detect ESCC and precancerous lesions in China.

Evaluation of Salivary Biomarkers and Spirometry for Diagnosing COPD in Non-Smokers and Smokers of Polish Origin.

Chronic obstructive pulmonary disease (COPD) is a prevalent respiratory condition with global implications. Accurate and timely diagnosis is critical; however, traditional diagnostic methods (based on spirometry) show limitations, prompting the search for predictive biomarkers and modern diagnostic techniques. This study explored the validation of COPD-related biomarkers (C-reactive protein, procalcitonin, neutrophil elastase, and alpha-1 antitrypsin) in saliva. A diverse cohort, including healthy non-smokers, healthy smokers, and COPD patients of Polish origin, underwent spirometry and marker analysis. The data correlated with clinical factors, revealing noteworthy relations. Firstly, salivary biomarker levels were compared with serum concentrations, demonstrating notable positive or negative correlations, depending on the factor. Further analysis within healthy individuals revealed associations between biomarker levels, spirometry, and clinical characteristics such as age, sex, and BMI. Next, COPD patients exhibited an enhanced concentration of biomarkers compared to healthy groups. Finally, the study introduced a breathing assessment survey, unveiling significant associations between self-perceived breathing and spirometric and tested parameters. Outcomes emphasized the relevance of subjective experiences in COPD research. In conclusion, this research underscored the potential of salivary biomarkers as diagnostic tools for COPD, offering a non-invasive and accessible alternative to traditional methods. The findings paved the way for improved modern diagnostic approaches.

Hyposalivation in patients with psoriasis: Association with severity, inflammatory, and anti-inflammatory cytokine biomarkers of the disease.

The aim of the study is to expound the effect of psoriasis on salivary glands by evaluating the secretion of saliva and salivary cytokine biomarkers in patients with psoriasis. This study was conducted by recruiting 120 subjects that included 60 patients diagnosed clinically with active psoriasis and 60 healthy controls who were age and gender matched to psoriatic subjects. Unstimulated whole saliva was collected from all the subjects by spitting method, and levels of tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin-2 (IL-2), and IL-10 (IL-10) were determined via enzyme-linked immunosorbent assay (BT Lab, Shanghai, China). Secretion of saliva in psoriasis patients was considerably reduced than in healthy controls. The concentrations of pro-inflammatory cytokines (TNF-α, IFN-γ, and IL-2) were significantly increased, whereas level of anti-inflammatory cytokine (IL-10) was markedly decreased in the saliva of psoriasis patients with hyposalivation compared to healthy subjects. Our results demonstrated significant negative correlation of salivary flow rates with the disease severity. No significant correlations were obtained between salivary levels of tested cytokines and salivary flow rates in our study. Findings of the study reflect inflammation of salivary glands with reduced salivary flow rates in psoriasis patients. The inflammatory responses in salivary gland tissues by virtue of increased pro-inflammatory cytokines concentrations together with lower anti-inflammatory cytokine levels may have a role in affecting the saliva secretion in psoriasis patients. Secretion of unstimulated saliva in psoriasis patients decreases with the severity and duration of the disease.

Salivary and serum haptoglobin, adenosine deaminase, and immunoglobulin G in growing pigs

BackgroundIdentification of animals in need of medical treatment is important in porcine health management, where analytical samples applicable at farm level could be utilized. Several biomarkers are measurable in saliva, which is less stressful to collect than blood. Saliva sampling is easy to learn and repeatable, making it suitable for monitoring purposes. Previous research suggests that porcine health biomarkers are dependent on production stage and gender, and that combining biomarkers improves diagnostic sensitivity. However, proper monitoring of biomarkers during the complete production cycle has not been studied. We aimed to describe the dynamics of salivary and serum haptoglobin (Hp), adenosine deaminase (ADA), and immunoglobulin G (IgG) in four production stages (suckling, early growing, late growing, finishing), on commercial Finnish pig farms using a total of 117 piglets. The relationship between gender and biomarker dynamics was investigated, as well as the relationships between these biomarkers in saliva and serum.ResultsThe highest salivary concentrations of Hp, ADA and IgG were measured in suckling piglets. The differences between production stages were generally larger in saliva than for the corresponding serum biomarkers. All correlation coefficients between salivary biomarkers were positive in each production stage and the strength of the correlation varied from 0.245 to 0.762. No similar trend was observed regarding correlation coefficients either between serum biomarkers or between salivary and serum biomarkers. Gender was associated with some biomarker concentrations.ConclusionsThe biomarker dynamics supported previous findings that collection of analytical samples should be conducted in age-matched populations. Positive and even strong relationships between salivary biomarkers indicate the potential to use especially saliva for health monitoring. Our results also suggest the importance of considering gender effects when assessing some salivary or serum biomarkers.

A Human Saliva Quality and Holistic Practices A Review on the Effects of Meditation, Dance and Exercise

This review article provides an overview of the effects of meditation, dance, and exercise on human saliva quality. Physical and mental fitness is essential for overall well-being, and engaging in these activities has been shown to promote physical and mental health. However, with the decline in physical activity due to technological advancements, stress has become a prevalent issue in modern society. Meditation, dance, and exercise have emerged as effective strategies for stress management. Stress levels can be measured through biomarkers, including those found in blood and saliva. Saliva may serve as a potential indicator of stress levels. Saliva contains various components that can serve as biomarkers for detecting systemic diseases and assessing overall health. Exploring the changes in saliva composition and understanding the physiological significance in response to meditation, dance, and exercise can provide valuable insights into the mechanisms underlying their positive effects on health. Saliva biomarkers have been associated with stress, inflammation, immune function, and even neurodegenerative disorders. Therefore, investigating the impact of these practices on saliva quality can uncover novel connections between mind-body interventions and overall well-being. This review highlights the importance of considering saliva as a valuable biofluid and encourages further research on salivary biomarkers in the context of meditation, dance, and exercise to expand our understanding of their therapeutic potential and optimise health outcomes.

Probing the salivary proteome for prognostic biomarkers in response to non-surgical periodontal therapy.

This prospective study investigated the salivary proteome before and after periodontal therapy. Ten systemically healthy, non-smoking, stage III, grade C periodontitis patients underwent non-surgical periodontal treatment. Full-mouth periodontal parameters were measured, and saliva (n = 30) collected pre- (T0), and one (T1) and six (T6) months post-treatment. The proteome was investigated by label-free quantitative proteomics. Protein expression changes were modelled over time, with significant protein regulation considered at false discovery rate <0.05. Treatment significantly reduced bleeding scores, percentages of sites with pocket depth ≥5 mm, plaque and gingival indexes. One thousand seven hundred and thirteen proteins were identified and 838 proteins (human = 757, bacterial = 81) quantified (≥2 peptides). At T1, 80 (T1 vs. T0: 60↑:20↓), and at T6, 118 human proteins (T6 vs. T0: 67↑:51↓) were regulated. The salivary proteome at T6 versus T1 remained stable. Highest protein activity post- versus pre-treatment was observed for cellular movement and inflammatory response. The small proline-rich protein 3 (T1 vs. T0: 5.4-fold↑) and lymphocyte-specific protein 1 (T6 vs. T0: 4.6-fold↓) were the top regulated human proteins. Proteins from Neisseria mucosa and Treponema socranskii (T1 vs. T0: 8.0-fold↓, 4.9-fold↓) were down-regulated. Periodontal treatment reduced clinical disease parameters and these changes were reflected in the salivary proteome. This underscores the potential of utilizing saliva biomarkers as prognostic tools for monitoring treatment outcomes.

Application of Salivary Alpha-1 Antitrypsin in the Diagnosis of Rheumatoid Arthritis: A Pilot Study.

Background and Objective: Rheumatoid arthritis (RA) is an autoimmune disease in which joints are gradually destroyed. Early diagnosis and treatment before joint deformation or destruction is important. The detection of novel RA biomarkers in saliva may facilitate early detection of RA before disease onset. This study aimed to evaluate salivary concentration of α1-antitrypsin (A1AT) in healthy patients and those with RA, and to assess the diagnostic value of salivary A1AT. Materials and Methods: In total, 80 participants were included: 20 healthy participants, and 60 patients with RA. Saliva and serum samples were obtained from all the patients. Levels of A1AT and cytokines, including interleukin-1 beta (IL-1β), IL-6, and IL-10 in saliva and serum, were evaluated using an enzyme-linked immunosorbent assay kit and Luminex assay. Data were analyzed using SPSS for Windows. Results: There was a higher level of A1AT in the saliva of patients with RA (median: 2388.66 ng/mL) than that in healthy controls (1579.06 ng/mL). There was a positive mild-to-moderate accuracy (area under the curve: 0.57-0.85) of A1AT in saliva to diagnose RA. The cut-off level (ng/mL) of A1AT in saliva for detecting RA was 1689.0. Conclusions: The obtained data can promote the application of the measurements of A1AT in saliva to diagnose RA.

A MODERN NONINVASIVE METHODS OF DIAGNOSIS GENITAL ENDOMETRIOSIS ASSOCIATED WITH INFERTILITY

Endometriosis is a disease characterized by an overgrowth of morphologically and functionally endometrium-like tissue outside the uterine cavity. Early diagnosis of endometriosis is very di ffi cult. The gold standard for diagnosis is the surgical method – laparoscopy, performed for infertility, pelvic pain or the presence of pelvic masses. However, this approach is currently being reconsidered, and the search for endometriosis markers for earlier diagnosis of this disease is relevant. The purpose of the literature review was to summarize the current data on promising noninvasive markers of endometriosis determined in various biological media. Materials and methods – summarized a modern data abot perspective noninvasive markes of genital endometriosis associated with infertility, determined diff erence biological fl uid. Results We evaluated more than 30 biomarkers in peritoneal fl uid, serum, urine, and saliva, as well as their combinations in publications that met the selection criteria. Studies have evaluated the diagnostic value and e ffi cacy of endometriosis biomarkers, but the results of these studies have sometimes been inconsistent. We were unable to identify a single biomarker or combination of biomarkers that was unequivocally clinically useful. The most signifi cant changes in biochemical composition were observed in peritoneal fl uid, but it requires invasive intervention to obtain it. Discussion. Saliva and urine studies have shown promising results in terms of diagnostic accuracy, but the evidence was of low quality for introduction into clinical guidelines. A number of serum biomarkers may be useful either for detecting endometriosis at early stages or for diff erentiating ovarian endometrioma from other benign ovarian masses, but the evidence for meaningful conclusions is insu fficient. Conclusion. None of the biomarkers have shown suffi cient accuracy for clinical use outside of the research environment, yet research in this area remains promising.

Exploration of reference genes for the development of a diagnostic kit on vascular aging in human saliva.

Identifying reliable biomarkers in saliva can be a promising approach to developing a rapid diagnostic kit for detecting vascular aging. This study investigated the most suitable reference gene for polymerase chain reaction (PCR) in saliva that is not affected by vascular aging variables. Whole saliva samples were collected to assess the expression of reference genes: actin beta (ACTB), 18S ribosomal RNA (18S rRNA), beta-2-microglobulin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The most abundantly expressed gene was 18S rRNA, and the least expressed gene was GAPDH. Four genes were ranked according to their relative stability, as determined by mathematical algorithms, indicating that ACTB and 18S rRNA were stably expressed as reference genes. 18S rRNA was identified as the most promising reference gene for detecting systemic diseases using saliva from patients with vascular aging in these limited experimental conditions.

Serum and salivary inflammatory biomarkers in juvenile idiopathic arthritis—an explorative cross-sectional study

BackgroundBiomarkers may be useful in monitoring disease activity in juvenile idiopathic arthritis (JIA). With new treatment options and treatment goals in JIA, there is an urgent need for more sensitive and responsive biomarkers.ObjectiveWe aimed to investigate the patterns of 92 inflammation-related biomarkers in serum and saliva in a group of Norwegian children and adolescents with JIA and controls and in active and inactive JIA. In addition, we explored whether treatment with tumor necrosis factor inhibitors (TNFi) affected the biomarker levels.MethodsThis explorative, cross-sectional study comprised a subset of children and adolescents with non-systemic JIA and matched controls from the Norwegian juvenile idiopathic arthritis study (NorJIA Study). The JIA group included individuals with clinically active or inactive JIA. Serum and unstimulated saliva were analyzed using a multiplex assay of 92 inflammation-related biomarkers. Welch’s t-test and Mann–Whitney U-test were used to analyze the differences in biomarker levels between JIA and controls and between active and inactive disease.ResultsWe included 42 participants with JIA and 30 controls, predominantly females, with a median age of 14 years. Of the 92 biomarkers, 87 were detected in serum, 73 in saliva, and 71 in both biofluids. A pronounced difference between serum and salivary biomarker patterns was found. Most biomarkers had higher levels in serum and lower levels in saliva in JIA versus controls, and in active versus inactive disease. In serum, TNF and S100A12 levels were notably higher in JIA and active disease. The TNF increase was less pronounced when excluding TNFi-treated individuals. In saliva, several biomarkers from the chemokine family were distinctly lower in the JIA group, and levels were even lower in active disease.ConclusionIn this explorative study, the serum and salivary biomarker patterns differed markedly, suggesting that saliva may not be a suitable substitute for serum when assessing systemic inflammation in JIA. Increased TNF levels in serum may not be a reliable biomarker for inflammatory activity in TNFi-treated children and adolescents with JIA. The lower levels of chemokines in saliva in JIA compared to controls and in active compared to inactive disease, warrant further investigation.

A systematic review of salivary biomarkers in Parkinson's disease.

The search for reliable and easily accessible biomarkers in Parkinson's disease is receiving a growing emphasis, to detect neurodegeneration from the prodromal phase and to enforce disease-modifying therapies. Despite the need for non-invasively accessible biomarkers, the majority of the studies have pointed to cerebrospinal fluid or peripheral biopsies biomarkers, which require invasive collection procedures. Saliva represents an easily accessible biofluid and an incredibly wide source of molecular biomarkers. In the present study, after presenting the morphological and biological bases for looking at saliva in the search of biomarkers for Parkinson's disease, we systematically reviewed the results achieved so far in the saliva of different cohorts of Parkinson's disease patients. A comprehensive literature search on PubMed and SCOPUS led to the discovery of 289 articles. After screening and exclusion, 34 relevant articles were derived for systematic review. Alpha-synuclein, the histopathological hallmark of Parkinson's disease, has been the most investigated Parkinson's disease biomarker in saliva, with oligomeric alpha-synuclein consistently found increased in Parkinson's disease patients in comparison to healthy controls, while conflicting results have been reported regarding the levels of total alpha-synuclein and phosphorylated alpha-synuclein, and few studies described an increased oligomeric alpha-synuclein/total alpha-synuclein ratio in Parkinson's disease. Beyond alpha-synuclein, other biomarkers targeting different molecular pathways have been explored in the saliva of Parkinson's disease patients: total tau, phosphorylated tau, amyloid-β1-42 (pathological protein aggregation biomarkers); DJ-1, heme-oxygenase-1, metabolites (altered energy homeostasis biomarkers); MAPLC-3beta (aberrant proteostasis biomarker); cortisol, tumor necrosis factor-alpha (inflammation biomarkers); DNA methylation, miRNA (DNA/RNA defects biomarkers); acetylcholinesterase activity (synaptic and neuronal network dysfunction biomarkers); Raman spectra, proteome, and caffeine. Despite a few studies investigating biomarkers targeting molecular pathways different from alpha-synuclein in Parkinson's disease, these results should be replicated and observed in studies on larger cohorts, considering the potential role of these biomarkers in determining the molecular variance among Parkinson's disease subtypes. Although the need for standardization in sample collection and processing, salivary-based biomarkers studies have reported encouraging results, calling for large-scale longitudinal studies and multicentric assessments, given the great molecular potentials and the non-invasive accessibility of saliva.

Salivary and serum biomarkers to evaluate psychological disorders in burning mouth syndrome: A systematic review and meta-analysis.

Burning mouth syndrome is a chronic pain syndrome mainly characterized by an intensive burning sensation of tongue. Previous studies have suggested that saliva/serum biomarkers in burning mouth syndrome might be associated with psychological disorders. The aim of systematic review was to observe whether the biomarkers in serum/saliva could be an alternative method to evaluate the psychological disorders in patients with burning mouth syndrome. The PubMed, Embase, and Cochrane Library databases were searched for papers published up to March 15, 2023. Risk of bias was measured by using the Newcastle-Ottawa Scale. RevMan was used for meta-analysis. A total of 467 articles were screened, which of 12 studies were included. These studies collected 43 different biomarkers in saliva and 35 in serum. Of these biomarkers, only three (cortisol, α-amylase, and IL-6) were analyzed in two or more studies. Only salivary cortisol levels were significantly higher in the patient group compared to the controls (Mean Difference = 1.39; 95% CI [0.80-1.97]; p < 0.001). Moreover, cortisol might be relevant to psychological scores, especially anxiety. Different papers have investigated salivary and serum biomarkers in burning mouth syndrome patients with controversial results. This meta-analysis showed that cortisol levels in saliva may be a potential biomarker to assess the psychological disorders in burning mouth syndrome patients.

Redox biomarkers in saliva and nuclear abnormalities in jugal epithelial cells of individuals with type 2 diabetes mellitus and periodontitis

ObjectiveTo evaluate salivary redox biomarkers levels in individuals with periodontitis and type 2 diabetes mellitus (T2DM) and correlate with periodontal parameters and nuclear alterations in epithelial cells from jugal mucosa. Design: Sixty individuals were categorized into three groups: T2DM with periodontitis (DM, n = 20), non-T2DM with periodontitis (PE, n = 20), and non-T2DM with periodontal health (HC, n = 20). All participants underwent fasting blood glucose and glycated hemoglobin measurements. After a periodontal examination, samples of epithelial cells from the jugal mucosa and saliva were collected. DNA damage was assessed by counting nuclear abnormalities using cytological analysis. Biomarkers of oxidative stress were determined through biochemical methods. Significant differences among groups were assessed using Kruskal-Wallis, Mann-Whitney, and Chi-square tests at a 5% significance level. Data were analyzed using Spearman’s correlation coefficient, linear regression, and logistic regression. ResultsFrequencies of nuclear abnormalities, as well as levels of reduced glutathione and uric acid, were significantly higher in the DM group compared to the PE and HC groups (p < 0.05). Fasting glucose, glycated hemoglobin, nuclear abnormalities, reduced glutathione, and uric acid exhibited positive correlations with periodontal parameters (p < 0.05). Furthermore, reduced glutathione was associated with dental biofilm (OR = 1.027 [95% CI, 1.004–1.049]) and condensed chromatin (OR = 0.415 [95% CI, 0.196–0.878]). ConclusionsPeriodontitis and T2DM are correlated with nuclear abnormalities, as well as salivary reduced glutathione and uric acid levels. Moreover, a higher prevalence of teeth with dental biofilm increases the likelihood of elevated levels of reduced glutathione in saliva, while the presence of condensed chromatin decreases that likelihood.

Visualization of Work Stress and Workload Data on Nurses at Makassar City Hospital (a Study Using Saliva Biomarker Examination Approach)

Background: Health services are one of the duties and responsibilities of a nurse. Providing optimal service is a major contribution to patient recovery. The implementation of services performed by nurses is usually faced with various conditions that can trigger work stress. Work stress is one of the obstacles for nurses in providing health services. The emergence of work stress in nurses due to their work being in close contact with patients and other factors such as work location, various inpatient cases, labor shortages, forced overtime hours, and the attitude of the head of the ward can put pressure on nurses. Objective: To determine the visualization of work stress and workload data on nurses at Makassar City Hospital using the approach of examining salivary biomarkers. Research methods: This study uses a longitudinal study approach, namely conducting an analytical study that aims to follow the development of work stress events in nurses at Makassar City Hospital. The research will be carried out for 3 years, namely 2023 – 2025. The first year of this research, namely 2023, has started from August-September 2023. The research location was carried out in 2 (two) Makassar City Hospitals namely Labuang Baji Hospital and Daya General Hospital. The number of samples in this study was 100 people. The research instrument used was a questionnaire used to collect primary data related to the characteristics of the respondents and workload. The cocorometer tool is used to measure work stress through salivary biomarkers. This tool functions to measure stress levels based on α-amylase in the salivary glands. Data were analyzed using descriptive analysis using Microsoft Excel and SPSS 21. Conclusion: Visualization of stress data found that respondents at Labuang Baji Hospital had 42 people who did not experience stress. Likewise, respondents at Daya General Hospital found 46 people who did not experience stress. This is in accordance with workload data for nurses in both hospitals. There were 16 people at Labuang Baji Hospital who stated their workload was light. It's the same with nurses who are at the Daya General Hospital. Most nurses stated that their workload was light, namely 27 people. It is hoped that nurses will continue to be able to control themselves (coping) against the working conditions they face, manage their time well, get enough rest, self-management to minimize the risk of stress. Another thing that is also important to apply is a healthy lifestyle and exercise. Hospital management is expected to properly implement policies and strategies related to stress and fatigue management in the workplace.

Effects of repeated intramuscular injections on sow behaviour reactions and stress-related saliva biomarkers – A pilot study

Medicating large production animals, such as sows, individually can be challenging, especially when repeated treatments are needed. Intramuscular (i.m.) injections have been shown to be aversive, and an increased reaction could be expected over consecutive days of injections. The aim of this pilot study was to assess if daily repeated i.m. injections cause sows to react differently with time and if injection stress is reflected in increased stress- and pain-related saliva biomarkers. Additionally, the aim was to assess if oxytocin was related to the behaviour reaction of sows during injections. In total 37 sows were allocated randomly into four treatment groups: 1. Sham-injected (SHAM: 9 sows), 2. Control (CON: 11 sows), 3. Penicillin (PEN: 9 sows) and 4. Tetracyclin (TET: 8 sows). The reaction of the sows to injections over three days was subjectively assessed by two observers. Further, SHAM and TET sows were saliva sampled for stress-related biomarkers before and after injections. Treatment (p < 0.001) influenced the medication reaction. Pairwise comparisons showed that the reaction to SHAM treatment was less compared to CON (p = 0.002) and PEN treatments (p < 0.001), while reactions to SHAM did not differ from TET (p > 0.1). Reaction to PEN was higher than that to TET (p = 0.01), while PEN and CON did not differ from each other (p > 0.1). Day tended to affect medication reaction (p = 0.08). The reaction to all other treatments decreased numerically from day 1 to day 3, while this was not the case for the TET treatment. Changes in biomarkers were subtle. Oxytocin tended to increase after TET injection on day 3 (p = 0.05). For the SHAM treatment, some of the biomarkers (chromogranin A, alpha-amylase activity and total esterase activity) showed significant or numerically decreasing trends in POST vs PRE-treatment samples on both days. The oxytocin level before injections was higher in sows that did not react to the medication at all than in sows that showed any kind of reaction (p = 0.008). In conclusion, sows appeared to habituate to the handling and injection, depending on the level of discomfort caused by the injectable substance. Further, the results suggest a potential role of oxytocin in decreasing aversive reactions in sows to human handling.

Identification of saliva biomarkers as tools for detection of early-stage oral squamous cell carcinoma

Cancer of the flat squamous cells of the oral cavity is called Oral Squamous Cancer Cell (OSCC). OSCC is responsible for 95% of all oral cancers. In high-risk countries like Pakistan, OSCC has one of the highest mortality rates amongst the general demographic. With most OSCC cases being identified in later stages, the 5-year survival rates have remained as low as 50%. However, with early diagnosis, OSCC patients have been shown to have a better chance of surviving. To that effect, identifying potential biomarkers to act as diagnostic indicators for OSCC can help in early diagnosis of the disease. An emerging field for alternative detection of cancer are non-invasive methods that use saliva as a fluid for liquid biopsies. Saliva comprises proteomes, microbiomes, metabolomes, transcriptomes, and genomes that can be used as potential tools for the early detection of cancer. “Salivaomics” has established itself as a promising area for diagnostic and prognostic purposes. In this review paper, we perform a meta-analysis and a systematic literature review to catalogue biomarkers found in saliva that could potentially be used in the early diagnosis of OSCC.

Stress and sleep deprivation-related biomarkers in saliva in patients with retinitis pigmentosa.

Patients with Retinitis Pigmentosa (RP) commonly experience sleep-related issues and are susceptible to stress. Moreover, variatiaons in their vision are often linked to anxiety, stress and drowsiness, indicating that stress and sleep deprivation lead to a decline in vision, and vision improves when both are mitigated. The objective of this study was to investigate the utility of salivary biomarkers as biochemical indicators of anxiety and sleep deprivation in RP patients. Seventy-eight RP patients and 34 healthy controls were included in this observational study. Anxiety and sleep-quality questionnaires, a complete ophthalmological exam for severity grading and, the collection of salivary samples from participants were assessed for participants. The activity of biomarkers was estimated by ELISA, and statistical analysis was performed to determine associations between the parameters. Associations between underlying psychological factors, grade of disease severity, and biomarkers activity were also examined. Fifty-two (67%) of patients had a severe RP, and 26 (33%) had a mild-moderate grade. Fifty-eight (58,9%) patients reported severe levels of anxiety and 18 (23.,1%) a high level. Forty-six (59%) patients obtained pathological values in sleep-quality questionaries and 43 (55.1%) in sleepiness. Patients with RP exhibited significant differences in testosterone, cortisol, sTNFαRII, sIgA and melatonin as compared to controls and patients with a mild-moderate and advanced stage of disease showed greater differences. In covariate analysis, patients with a severe anxiety level also showed greater differences in mean salivary cortisol, sTNFαRII and melatonin and male patients showed lower IgA levels than female. The present findings suggest that salivary biomarkers could be suitable non-invasive biochemical markers for the objective assessment of sleep deprivation and anxiety in RP patients. Further research is needed to characterize the effects of untreated negative psychological states and sleep deprivation on increased variability of vision and disease progression, if any.

A salivary urea sensor based on a microsieve disposable gate AlGaN/GaN high electron mobility transistor.

The abundant bio-markers in saliva provide a new option for non-invasive testing. However, due to the presence of impurities in the saliva background, most of the existing saliva testing methods rely on pre-processing, which limits the application of saliva testing as a convenient means of testing in daily life. Herein, a disposable-gate AlGaN/GaN high electron mobility transistor (HEMT) biosensor integrated with a micro-sieve was introduced to solve the problem of signal interference caused by charged impurities in saliva for HEMT based biosensors, where the micro-sieve was utilized as a pre-treatment unit to remove large particles of impurities from saliva through the size effect and thus greatly improving the accuracy of detection. The experimental results showed that the HEMT based biosensor has excellent linearity (R2 = 0.9977) and a high sensitivity of 6.552 μA dec-1 for urea sensing from 1 fM to 100 mM in 0.1× PBS solution. When it comes to artificial saliva detection, compared to the HEMT sensor without the micro-sieve (sensitivity = 3.07432 μA dec-1), the sensitivity of the HEMT sensor integrated with the micro-sieve showed almost no change. Moreover, to verify that urea can be detected in actual saliva, urea is sensed directly in human saliva. The addition of the microsieve module provides a new way for biosensors to detect specific markers in saliva in real time, and the designed HEMT biosensor with the microsieve function has a wide range of application potential in rapid saliva detection.

Sputum production and salivary microbiome in COVID-19 patients reveals oral-lung axis.

SARS-CoV-2, a severe respiratory disease primarily targeting the lungs, was the leading cause of death worldwide during the pandemic. Understanding the interplay between the oral microbiome and inflammatory cytokines during acute infection is crucial for elucidating host immune responses. This study aimed to explore the relationship between the oral microbiome and cytokines in COVID-19 patients, particularly those with and without sputum production. Saliva and blood samples from 50 COVID-19 patients were subjected to 16S ribosomal RNA gene sequencing for oral microbiome analysis, and 65 saliva and serum cytokines were assessed using Luminex multiplex analysis. The Mann-Whitney test was used to compare cytokine levels between individuals with and without sputum production. Logistic regression machine learning models were employed to evaluate the predictive capability of oral microbiome, salivary, and blood biomarkers for sputum production. Significant differences were observed in the membership (Jaccard dissimilarity: p = 0.016) and abundance (PhILR dissimilarity: p = 0.048; metagenomeSeq) of salivary microbial communities between patients with and without sputum production. Seven bacterial genera, including Prevotella, Streptococcus, Actinomyces, Atopobium, Filifactor, Leptotrichia, and Selenomonas, were more prevalent in patients with sputum production (p<0.05, Fisher's exact test). Nine genera, including Prevotella, Megasphaera, Stomatobaculum, Selenomonas, Leptotrichia, Veillonella, Actinomyces, Atopobium, and Corynebacteria, were significantly more abundant in the sputum-producing group, while Lachnoanaerobaculum was more prevalent in the non-sputum-producing group (p<0.05, ANCOM-BC). Positive correlations were found between salivary IFN-gamma and Eotaxin2/CCL24 with sputum production, while negative correlations were noted with serum MCP3/CCL7, MIG/CXCL9, IL1 beta, and SCF (p<0.05, Mann-Whitney test). The machine learning model using only oral bacteria input outperformed the model that included all data: blood and saliva biomarkers, as well as clinical and demographic variables, in predicting sputum production in COVID-19 subjects. The performance metrics were as follows, comparing the model with only bacteria input versus the model with all input variables: precision (95% vs. 75%), recall (100% vs. 50%), F1-score (98% vs. 60%), and accuracy (82% vs. 66%).