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Background: In order to obtain sustainable analytical methods, it is essential to develop kinetically efficient sample preparation strategies in which equilibria are reached faster, as dispersive extraction techniques. In addition, the higher the reduction in size, the higher number of extraction vessels can be located and thus the higher number of samples can be simultaneously treated. All this increases sample throughput, and contributes to the reduction of chemical waste, and energy and sample consumption. In this sense, multiposition extraction platforms are smart strategies to achieve these goals, but they are scarcely developed for dispersive extraction techniques., Results: Taking miniaturized stir bar sorptive dispersive microextraction (mSBSDME) as a starting point, a 96-position extraction platform has been developed using a 96-position stirring plate and a tailor-designed 3D-printed support for locating the miniaturized extraction vessels, achieving a high-throughput miniaturized sample preparation strategy. In order to show the applicability of this novel platform, the determination of Δ 9 -tetrahydrocannabinol (THC) and cannabidiol (CBD) in human saliva has been carried out and applied to samples collected after the consumption of marijuana and legal CBD-rich cannabis. Only 100 μL of saliva were needed for the analysis and good analytical features in terms of linearity (at least up to 500 ng mL -1 ), limits of detection (0.7 and 2.8 ng mL -1 for THC and CBD, respectively), and precision (RSD ≤ 14 %) were achieved., Significance: The miniaturization of the vessel allows the use of small volumes of sample (i.e., a few microliters) and the treatment of 96 samples in parallel, being the first proposal for carrying out dispersive sorbent-based microextraction under the concept of 96-well format. Additionally, this new workflow contributes to the development of analytical methods that meet the three pillars of sustainability, i.e., greenness and easily affordable in terms of economics and applicability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Adolescent depression is a major public health concern. Although stress has been linked to more severe depression, its association with mild depression among adolescents is not understood. This study assesses the relationship between perceived stress and cortisol (a physiologic measure of stress) and examines the relationships between these stress measures and depressive symptoms among adolescents 13-19 years of age., Methods: Stress was measured with the Perceived Stress Scale-10 and through salivary sampling for cortisol four times throughout the day. The Patient Health Questionnaire-9 was used to measure depressive symptoms (range 0-27), where ≥5 indicated the threshold for experiencing at least mild depressive symptoms. Spearman coefficients and multiple logistic regression models were used to examine the relationships between our variables of interest., Results: The mean age of the 73 participants in our study was 15.82 years. 49 % of the participants reported depressive symptoms (PHQ-9 score ≥ 5). Both higher perceived stress (odds ratio [OR] = 1.11, p = 0.022) and lower cortisol (area-under-the curve; AUC G ) (OR = 0.99, p = 0.009) were associated with increased odds of having depressive symptoms., Limitations: Few participants had moderate to severe PHQ-9 depression, therefore our study reported findings on mild depression or greater., Conclusions: Perceived stress and cortisol appear to reflect distinct, independent components of the stress experience. However, both greater perceived stress and less circulating cortisol may indicate difficulties in regulating stress as potential factors underlying depressive symptoms. Future research should focus on the different types of adolescent stressors and the importance of routine screening of stress and depression, including mild depression., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This work presents a novel method for the analysis of polycyclic aromatic hydrocarbons (PAHs) in saliva samples using solid phase microextraction (SPME) coupled with gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS). The protocol utilizes the latest commercially available overcoated fiber (PDMS/DVB/PDMS) for direct immersion extraction of the target analytes, enabling the determination of thirteen PAHs, including low-volatile compounds. The SPME extraction method was optimized using a central composite design (CCD). The evaluation of the fiber coating's robustness over time demonstrated excellent extraction performance with no significant degradation. The validation procedure confirmed good performance for all parameters, with LOQ values (100 ng/L for ten analytes and 500 ng/L for three analytes) comparable to other chromatographic methods. The environmental impact of the protocol was objectively assessed using two recently proposed metrics: the Green Analytical Procedure Index (GAPI) and the Analytical Greenness metric for sample preparation (AGREEprep). Both metrics indicated good overall environmental friendliness, with AGREEprep providing a satisfactory comprehensive score despite the use of highly impactful instrumentation. These characteristics make the developed method suitable for routine analysis in environmental and epidemiological monitoring., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study investigated methods for sampling bile acids in saliva, a potential non-invasive diagnostic biofluid. Bile acids have been implicated in causing damage and permanent changes to the esophageal mucosa and increasing the risk of developing Barrett's esophagus, a condition that can potentially progress to esophageal cancer. Three saliva collection methods were compared: spitting, Salivette® swabs, and Salivette Cortisol® swabs. Spitting emerged as the superior method with the highest recoveries and the least interference, likely due to Salivette swabs retaining bile acids or introducing unknown interferences. All saliva samples were analyzed by UHPLC-MS/MS using the Zorbax RRHD Eclipse Plus C18 column (3 × 50 mm, 1.8 µm) in gradient elution of 0.1 % formic acid in water and methanol. Saliva sample stability was assessed over 14 days, reflecting typical storage times. The levels of detected bile acids were stable for the measured period (RSD ≤ 22 %) and no degradation was observed. Bile acid levels in saliva fluctuated throughout the day, with the greatest changes observed for glycine-conjugated bile acids after meals. To minimize sampling variability, saliva collection by spitting after overnight fasting is recommended for future studies. Our findings are applicable for standardized bile acid sampling and are currently applied in a large clinical study evaluating bile acids as potential susceptibility markers for Barrett's esophagus diagnostics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR) has been applied to determine salivary biomarkers with high sensitivity and cost-effectiveness. Our study aimed to test the hypothesis that the spectral profile of saliva demonstrates distinct vibrational modes corresponding to different exercise protocols, thereby facilitating exercise monitoring. Saliva samples were collected from trained male subjects at three intervals: pre-exercise, post-exercise, and 3 h post-exercise. The protocols included acute sessions of continuous exercise (CE), high-intensity interval exercise (HIIE), and resistance exercise (RE). ATR-FTIR analysis revealed that salivary biochemical components changed uniquely with each exercise protocol. Specific spectral vibrational modes were identified as potential biomarkers for each exercise type. Notably, the salivary spectrum pattern of CE closely resembled that of HIIE, whereas RE showed minor alterations. Furthermore, we attempted to apply an algorithm capable of distinguishing the spectral range that differentiates the exercise modalities. This pioneering study is the first to compare changes in saliva spectra following different exercise protocols and to suggest spectrum peaks of vibrational modes as markers for specific types of exercises. We emphasize that the spectral wavenumbers identified by FTIR could serve as practical markers in distinguishing between different exercise modalities, with sensitivity, specificity, and accuracy correlating with the metabolic changes induced by exercise. Therefore, this study contributes a panel of ATR-FTIR spectral wavenumbers that can be referenced as a spectral signature capable of distinguishing between resistance and endurance exercises., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Meconium, the first stool produced by neonates, has been used as an analyte for exogenous fetal exposures. However, few studies have investigated the relationship between meconium and androgen exposure in utero. Here, we examine the associations of testosterone and dehydroepiandrosterone (DHEA) across maternal antenatal salivary testosterone, cord blood, meconium, and infant salivary testosterone. A total of 47 women with singleton, uncomplicated pregnancies, and their infants were included in this study. Participants were recruited from an academic obstetric clinic. Maternal saliva was collected at 36-weeks' gestation. Cord blood and meconium were collected at birth. Infant salivary testosterone was collected at 1 and 4 weeks of age. Multivariate model results showed that meconium testosterone was associated with neonatal testosterone at 1 (F = 5.62, p = 0.029) and 4 weeks (F = 4.28, p = 0.048) postnatal age; no sex differences were detected. This study suggests meconium is a valuable tool for evaluating endogenous androgen exposure and should be used in future studies to investigate the fetal hormonal milieu., (© 2024 Wiley Periodicals LLC.)

Research and theory suggest an important role of neuroendocrine function in emotional development, particularly under conditions of elevated stress. We provide empirical data to clarify associations between alpha-amylase (AA) and cortisol as well as test the differential linkages among AA, cortisol, and symptoms of anxiety, depression, and posttraumatic stress in children. Children recruited from a low-income elevated violence community (n = 100; mean age = 10, SD = 0.64; 79% Latino; 67% received free or reduced lunch) were assessed on diurnal levels of AA and cortisol along with assessments of anxiety, depression, and posttraumatic stress symptoms (PTSS). Elevated anxiety symptoms were associated with steeper linear slopes of AA with higher levels of AA in the morning but lower levels of AA in the evening. Depression was associated with differential cubic trajectories of AA when PTSSs were included in the model. Anxiety also predicted differential cubic diurnal trends in cortisol, such that greater anxiety symptoms were associated with relatively higher levels of cortisol in the evening. Again, depression symptoms when PTSS were included predicted diurnal cubic trends with elevated depression associated with lower awakening and midday cortisol that reversed to higher evening cortisol compared to youth with fewer self-reported depression symptoms., (© 2024 The Author(s). Developmental Psychobiology published by Wiley Periodicals LLC.)

Background: Previous studies have revealed the importance of CYFRA 21-1 in the diagnosis of oral squamous cell cancer (OSCC). However, the results are inconsistent. This meta-analysis is to evaluate CYFRA 21-1's efficacy in distinguishing OSCC., Methods: Systematic searches of Web of Science, PubMed, and CNKI (1996-2024) were conducted following the Preferred Reporting ltems for Systematic Reviews and Meta-Analyses (PRISMA) guidelines., Results: Analysis of 693 patients and 548 controls yielded combined sensitivity (SEN) of 0.71 (95% CI: 0.68, 0.75), specificity (SPE) of 0.88 (95% CI: 0.85, 0.90), and area under the curve (AUC) of 0.927. Subgroup analysis showed higher SEN (0.88), SPE (0.93), and AUC (0.962) in saliva versus serum. Enzyme-linked immunosorbent assay (ELISA) demonstrated superior performance over electrochemiluminescent immunoassay (ECLIA) (AUC: 0.968 vs. 0.868)., Conclusion: CYFRA 21-1 is effective in OSCC diagnosis, with ELISA showing better sensitivity. Saliva emerges as a promising diagnostic medium compared to serum., Registration: PROSPERO CRD42024566835.

Background and Purpose: The definitive diagnosis of gastroesophageal reflux disease (GERD) often requires invasive investigations like upper gastrointestinal endoscopy or reflux monitoring. We aimed to explore the relationship between salivary pepsin and GERD and its value as a non-invasive diagnostic tool., Methods: Databases (PubMed, Web of Science, Cochran Library, and EMBASE) were searched from their inception to January 22, 2024 to explore the correlation of salivary pepsin with GERD. The meta-analysis data retrieved were summarized, including the salivary pepsin concentration, sensitivity of diagnosis (SEN), specificity of diagnosis (SPE), negative likelihood ratio, positive likelihood ratio, diagnostic odds ratio, and receiver operating characteristic (ROC) curve., Results: The meta-analysis comparing salivary pepsin concentration in two groups (proven GERD and non-GERD) with 18 studies revealed that the proven GERD group had higher salivary pepsin concentration than the non-GERD group (SMD = 1.74 [95% CI 1.14-2.34]). The meta-analysis of salivary pepsin diagnostic value for proven GERD incorporated 23 studies. The results showed pooled SEN (0.73 [95% CI 0.66-0.80]), SPE (0.72 [95% CI 0.65-0.78]), positive likelihood ratio (2.61 [95% CI 2.02-3.39]), negative likelihood ratio (0.37 [95% CI 0.28-0.50]), diagnostic odds ratio (7.03 [95% CI 4.24-11.66]) and area under the SROC curve (0.79 [95% CI 0.75-0.82])., Conclusion: GERD patients presented a higher salivary pepsin concentration. Salivary pepsin is both sensitive and specific in identifying GERD, making it a promising non-invasive marker for diagnosis., (© 2024 John Wiley & Sons Ltd.)

Introduction: The association between dental caries, salivary alterations, and type 2 diabetes (T2D) is inconclusive. This systematic review evaluated the caries status and salivary alterations in adults with T2D., Data: Observational studies (cross-sectional, case-control, or cohort studies) that evaluated the association between type 2 diabetes and dental caries and/or salivary changes were included., Sources: Systematic searches of PubMed/Medline, Web of Science, Embase, and the Cochrane Library for relevant literature published up to January 2024 were performed., Study Selection: Ninety-eight studies were selected from the 1981 records obtained after screening the databases. Thirty-nine studies that met the eligibility criteria were included in the qualitative synthesis, and twenty-eight of the included studies were suitable for the quantitative synthesis (meta-analysis)., Results: The meta-analysis showed that compared to non-diabetic individuals, type 2 diabetic individuals had higher DMFT (mean difference: 2.27; 95 % CI, 1.31 to 3.22; P < 0.00001). Moreover, type 2 diabetic individuals had lower salivary flow rate (standardized mean difference: -2.62; 95 % CI, -4.38 to -0.86; P = 0.003), salivary pH (standardized mean difference: -1.35; 95 % CI, -1.89 to -0.80; P < 0.00001) and salivary buffering capacity (standardized mean difference: -0.60; 95 % CI, -0.93 to -0.27; P = 0.0004). Whereas DMFT (mean difference: -2.15; 95 % CI, -6.47 to 2.17; P = 0.33) and salivary buffering capacity (standardized mean difference: -0.05; 95 % CI, -0.48 to 0.37; P = 0.80) were not significantly different between individuals with well-controlled diabetes and those with poorly-controlled diabetes., Conclusions: Type 2 diabetes may lead to an increased index of dental caries and a reduction in salivary flow rate, salivary pH, and salivary buffering capacity in adults., Clinical Significance: Individuals with type 2 diabetes exhibit an elevated risk of dental caries and salivary alterations that facilitate the progression of dental caries. It is recommended that dentists and endocrinologists focus their attention on the oral health of this population., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objective: To investigate the association between interleukin-8 (IL-8) levels in gingival crevicular fluid (GCF) and total oral fluid (TOF) and the responsiveness to steps 1 and 2 of periodontal therapy., Materials and Methods: One-hundred and fifty-nine patients affected by periodontitis received steps 1 and 2 of periodontal therapy. At baseline, TOF and GCF samples were collected and analysed for IL-8 (Il-8 TOF /IL-8 GCF ) using flow cytometry. Therapy outcomes were relative proportions of residual periodontal pockets (PPD%), pocket closure (PC) rates and pocket probing depth (PPD) reductions; these were associated with IL-8 TOF /IL-8 GCF ., Results: High IL-8 TOF was significantly associated with higher residual PPD% (p = 0.044) and lower PPD reduction compared to low IL-8 TOF (high 0.79 ± 1.20 mm vs. low 1.20 ± 1.20 mm, p < 0.001) in non-smokers, while in smokers high IL-8 GCF was related to lower PPD reduction (high 0.62 ± 1.22 mm vs. low 0.84 ± 1.12 mm, p = 0.009). Furthermore, high baseline IL-8 TOF was significantly associated with poorer PC rates compared to medium and low concentrations in both non-smokers (high 41% vs. medium 55% vs. low 58%, p < 0.001) and smokers (high 34% vs. medium 44% vs. low 46%, p < 0.001)., Conclusion: High IL-8 concentrations at baseline had a significant impact on residual PPD%, PC rates and PPD reduction. The findings suggest that, especially in non-smokers, baseline IL-8 levels collected from the TOF could serve as a component in the estimation of responsiveness to steps 1 and 2 of periodontal therapy., (© 2024 The Author(s). Journal of Clinical Periodontology published by John Wiley & Sons Ltd.)

Oxytocin pathways are hypothesized to play important roles in human-animal interactions and may contribute to some benefits of these interspecific social relationships. We explored the effects of naturalistic interactions between children and dogs on oxytocin release in both species, as well as associations between methylation of the oxytocin receptor gene (OXTRm), social behavior, and oxytocin response in this context. Children (N = 55) participated in a within-subjects design involving a) interaction with their pet dog, b) interaction with an unfamiliar dog, and c) a nonsocial control condition (solitary play). We used immunoassays to measure salivary and urinary oxytocin in both the children and dogs, behavioral coding to characterize dog-child interactions, and bisulfite sequencing to quantify methylation of the oxytocin receptor gene (N = 32 children). Child salivary oxytocin decreased moderately across time in all conditions, but the extent of this effect varied between conditions, with greater oxytocin output during interactions with dogs than the control condition. In the pet dog condition, children's salivary oxytocin response was positively associated with the duration of visual co-orientation between the child and dog. Child urinary oxytocin did not deviate substantially from baseline in any condition. Children with higher levels of OXTRm had greater oxytocin output during interactions with their pet dogs, but lower oxytocin output in the control condition, and engaged in lower levels of affectionate interaction with dogs across conditions. Children's pet dogs exhibited increases in salivary oxytocin, but we observed the opposite pattern in the unfamiliar dog, who exhibited decreases in both urinary and salivary oxytocin on average. Collectively, our results support the hypothesis that oxytocin pathways may shape and respond to social interactions between children and dogs, highlighting an important role for companion animals in child development., Competing Interests: Declaration of Competing Interest None., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Difficulties in fear regulation can sometimes result in maladaptive fear responses. To better understand how to improve fear regulation, it is important to determine how known factors, such as sex hormone status and stress, might interact to influence fear memory. Research has shown that women with high estradiol levels (mid-cycle) and men exhibit better extinction retention compared to women with low estradiol levels (women in the early follicular cycle or using oral contraceptives). Stress has also been demonstrated to affect both the learning and retention of extinction. Despite documented interactions between stress and sex hormones, their combined effects have not been thoroughly studied. This study aims to examine the impact of stress as a function of sex hormone status on extinction learning and retention. A total of 168 non-clinical participants were studied, including men (n = 46), women using oral contraceptives (n = 38), women in the early follicular phase (n = 40), and women in mid-cycle (n = 44). On Day 1, fear acquisition training was performed. On day 2, prior to extinction training, half of the participants were exposed to a psychosocial stressor, while the other half performed a non-stressful control task. On day 3, extinction retention was tested. Fear was quantified using skin conductance responses, while stress hormones were quantified through saliva samples. Exposure to stress prior to extinction training did not affect extinction learning, regardless of sex hormone status. In contrast, pre-extinction stress exposure had different effects on extinction retention depending on hormone status. Stressed men showed impairment in extinction retention compared to controls, while the experimental condition had no effect on naturally cycling women. Regardless of stress exposure, early follicular women exhibited a deficit in fear regulation, while mid-cycle women showed effective fear regulation. Among women using oral contraceptives, the stress group demonstrated better extinction retention compared to the control group. These results demonstrate the importance of considering sex hormone status and stress exposure during extinction learning, as both components may modulate extinction retention. These results could help identifying hormonal conditions that may enhance the effectiveness of extinction-based psychological therapies used in the treatment of fear-related disorders., Competing Interests: Declaration of Competing Interest none, (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The inference of body fluids and tissues is critical in reconstructing crime scenes and inferring criminal behaviors. Nevertheless, present methods are incompatible with conventional DNA genotyping, and additional testing might result in excessive consumption of forensic scene materials. This study aims to investigate the feasibility of distinguishing common body fluids/tissues through the difference in mitochondrial DNA copy number (mtDNAcn). Four types of body fluids/tissues were analyzed in this study - hair, saliva, semen, and skeletal muscle. MtDNAcn was estimated by dividing the read counts of mitochondrial DNA to that of nuclear DNA (RR mt/nu ). Results indicated that there were significant differences in RR mt/nu between different body fluids/tissues. Specifically, hair samples exhibited the highest RR mt/nu (log 10 RR mt/nu : 4.3 ± 0.28), while semen samples showed the lowest RR mt/nu (log 10 RR mt/nu : -0.1 ± 0.28). RR mt/nu values for DNA samples without extraction were notably higher (approximately 2.9 times) than those obtained after extraction. However, no significant difference in RR mt/nu was observed between various age and gender groups. Hierarchical clustering and Kmeans clustering analyses showed that body fluids/tissues of the same type clustered closely to each other and could be inferred with high accuracy. In conclusion, this study demonstrated that the simultaneous detection of nuclear and mitochondrial DNA made it possible to perform conventional DNA analyses and body fluid/tissue inference at the same time, thus killing two birds with one stone. Furthermore, mtDNAcn has the potential to serve as a novel and promising biomarker for the identification of body fluids/tissues., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Situational factors can increase people's vulnerability to intergroup bias, including prejudicial attitudes, negative stereotyping, and discrimination. We proposed that increases in inflammatory activity that coincide with acute illness may represent a hitherto unstudied situational factor that increases intergroup bias. The current study experimentally manipulated increases in inflammatory activity by administering the seasonal influenza vaccine or a saline placebo. We quantified inflammatory activity by assessing change in salivary pro-inflammatory cytokines and assessed intergroup bias using a resume evaluation task and self-reported ethnocentrism. Primary analyses focused on a subsample of 117 participants who provided high quality data; robustness analyses included various permutations of lower quality participants. Findings revealed that changes in the cytokine interleukin-1β (IL-1β) in response to the vaccine were associated with greater intergroup bias. Among participants who received the vaccine, IL-1β change was negatively associated with evaluation of a Latina (but not a White woman) applicant's competency and recommended starting salary. Moreover, IL-1β change was positively associated with ethnocentrism. Overall, results provide support for the hypothesis that acute illness, via the mechanistic role of inflammatory cytokines, affects social cognition in ways that can increase intergroup bias., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Determining the source of body fluids is crucial in forensic investigations, as it provides valuable information about suspects and the nature of the crime. Microbial markers that trace the source of tissues and body fluids based on site specificity and temporal stability are often used effectively for this purpose. In this study, a multiplex system comprising seven microbial markers (Finegoldia magna, Corynebacterium tuberculostearicum, Cutibacterium acnes, Haemophilus parainfluenzae, Streptococcus oralis, Prevotella melaninogenica and Faecalibacterium prausnitzii) was developed to distinguish between skin, saliva, and feces samples. Based on these markers, the system produces electropherograms that are specific for each sample type. We collected 492 samples from six different skin sites (palm, antecubital crease, inguinal crease, cheek, upper back, and toe web space), the buccal mucosa, and stool were collected to further test the system. Beta diversity analysis revealed distinct clustering among the three sample groups. Additionally, skin microenvironment cluster analysis was used to identify skin sites accurately. This analysis classified skin samples into four distinct microenvironments: dry, moist, oily, and foot. Finally, we established a machine learning prediction model based on random forest regression to identify the skin microenvironment, achieving an overall prediction accuracy of 79 %. The multiplex system developed in this study accurately identifies the sources of body fluids, and the skin microenvironment. These findings offer new insights into the application of microbial markers in forensic science., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Small molecules with enzyme-like properties have recently attracted considerable attention. Herein, we discovered that nitrite possesses intrinsic oxidase-mimicking activity upon visible light, catalyzing the oxidation of the typical chromogenic substrate in the absence of H 2 O 2 . Notably, nitrite exhibited a markedly high value of K cat , approximately 4, 7, and 4000-fold greater than that of Acr + -Mes, Eosin Y, and Diacetyl, respectively. Comprehensive investigation elucidated that O 2 • ⁻ and •OH are the primary reactive oxygen species (ROS) responsible for the oxidation of 3,3',5,5'-tetramethylbenzidine dihydrochloride hydrate (TMB). Leveraging the linear correlation between the absorbance of oxidized TMB (oxTMB) at 652 nm and nitrite concentration, a simple colorimetric approach for nitrite detection was successfully established in the range of 1-75 μM with a detection limit of 0.14 μM. Moreover, the proposed strategy could be applied to determine the nitrite concentration in saliva, exhibiting a great prospect for clinical diagnosis. This work contributes novel insights into the exploration of small-molecule enzyme mimics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In the past few years, the COVID-19 pandemic, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) seriously threatens global public health security due to its high contagiousness. It remains of vital importance to develop a rapid and sensitive assay for SARS-CoV-2. In this work, we proposed a sandwich-type assay based on poly(N-isopropylacrylamide) (PNIPAM), allowing efficient detection of the SARS-CoV-2 S1 protein in the homogeneous solution. Firstly, a direct sandwich-type assay was established with a linear range of 0.2-2 μg/mL and a limit of detection (LOD) of 0.11 μg/mL, which could realize rapid detection in about 1 h. Furthermore, the sandwich-type assay coupled with rolling circle amplification (RCA) obtained an increase in sensitivity of 5.9 × 10 4 folds with a wide linear range of 0.01 - 100 ng/mL and a LOD of 1.88 pg/mL. The average recoveries in unpretreated saliva were 90 %-113.0 %, indicating the potential of the developed method for application in practical samples. Given the high selectivity and sensitivity of the developed method, it has a significant potential for rapid and early detection of SARS-CoV-2., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Halitosis in children implies psychosocial repercussions. Risk factors associated with this condition are unclear, and detection methods are inaccurate., Aim: To quantify the levels of sulfur-like compounds in children with asthma and healthy children from a novel validated assay, and to establish the risk factors related to halitosis., Design: One hundred and twenty-eight individuals (63 healthy and 65 asthmatic) from 3 to 17 years of age were tested using a passive colorimetric sensor to measure the levels of sulfur-like compounds in breath and saliva. Information was collected on oral hygiene habits, gingival and dental health, breathing type, and dental malocclusion., Results: The mean values of hydrogen sulfide were 4.0 ± 6.8 and 19.7 ± 12.2 ppbv (parts per billion in volume) in the control and asthmatic groups, respectively (p < .001). The presence of higher concentrations of sulfur compounds was significantly associated (p < .05) with the presence of gingival inflammation, tongue coating, dental plaque, mouth breathing, hypomineralization, age, tongue brushing, and the use of dental floss., Conclusion: The level of sulfur in breath and saliva was significantly higher in patients with asthma. These results can serve as a precedent to raise awareness among paediatricians and parents about oral hygiene care in children and adolescents., (© 2024 The Authors. International Journal of Paediatric Dentistry published by BSPD, IAPD and John Wiley & Sons Ltd.)

Forensic genetic analyses aim to retrieve as much information as possible from biological trace material recovered from crime scenes. While standard short tandem repeat (STR) profiling is essential to individualize biological traces, its significance is diminished in crime scenarios where the presence of a suspect's DNA is acknowledged by all parties. In such cases, forensic (m)RNA analysis can provide crucial contextualizing information on the source level about a trace's composition, i.e., body fluids/tissues, and has therefore emerged as a powerful tool for modern forensic investigations. However, the question which of several suspects contributed a specific component (body fluid) to a mixed trace cannot be answered by RNA analysis using conventional methods. This individualizing information is stored within the sequence of the mRNA transcripts. Massively parallel sequencing (MPS) represents a promising alternative, offering not only higher multiplex capacity, but also the typing of individual coding region SNPs (cSNPs) to enable the assignment of contributors to mixture components, thereby reducing the risk of association fallacies. Herein, we describe the development of an extensive mRNA/cSNP panel for targeted sequencing on the IonTorrent S5 platform. Our panel comprises 30 markers for the detection of six body fluids/tissues (blood, saliva, semen, skin, vaginal and menstrual secretion), along with 70 linkage-controlled cSNPs for contributor assignment. It exhibited high reliable detection sensitivity with RNA inputs down to 0.75 ng and a conservatively calculated probability of identity of 0.03 - 6 % for individual body fluid-specific cSNP profiles. Limitations and areas for future work include RNA-related allele imbalances, inclusion of markers to correctly identify rectal mucosa and the optimization of specific markers. In summary, our new panel is intended to be a major step forward to interpret biological evidence at sub-source and source level based on cSNP attribution of a body fluid component to a suspect and victim, respectively., Competing Interests: Declaration of Competing Interest All authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Home testing technology strategy is critical for early screening of disease. However, current home testing technologies often require complex processes, which limits their application. In this study, a time-resolved cascade logic gate microfluidic chip (TCLMC) was revealed to enable capillary force-based one-step operation without manual intervention or professional equipment. By analogy with logic gates in the circuit, TCLMC could automatically control the fluid flow and regulate the incubation time to optimize the immunoassay. The limit of detection of TCLMC for SARS-CoV-2 and influenza B virus (Flu B) was 134.94 and 79.17 pg mL -1 within 10 min. Additionally, this study tested saliva samples from 12 Flu B patients and 24 healthy controls to verify its clinical application. The results showed that TCLMC had high sensitivity (100%), specificity (100%), and accuracy (100%). This study provides a new one-step strategy for home testing and demonstrates its great potential in the diagnosis field., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Objectives: To investigate the effects of a smartphone-based oral and whole-body exercise programme on oral function in older adults., Methods: Individuals aged 65 years or above were randomized into three groups (non-app use, app use and control group), and a combined oral and whole-body exercise programme was conducted for 5 weeks. Oral muscle strength, saliva flow rate, Oral Health Impact Profile-14 (OHIP-14), and Geriatric Oral Health Assessment Index (GOHAI) were measured before and after the intervention. The changes in each group were analysed at the end of the programme using paired sample t-tests, and the differences among the groups were analysed using the chi-square test, Fisher's exact test and ANOVA., Results: The anterior tongue strength increased by 2.80 kPa after the intervention in the non-app use group; however, the change was not statistically significant. In the app use group, the anterior tongue strength significantly increased by 4.48 kPa. The saliva flow rate increased by 0.54 and 0.71 g/min in the non-app and app use groups, respectively, after the intervention (p < 0.05), and the change was greater in the app use group than in the other groups (p < 0.01). There were no significant changes in the posterior tongue strength, cheek strength, OHIP-14 or GOHAI scores over the course of the study., Conclusions: A smartphone-based combined oral and whole-body exercise programme can improve anterior tongue strength and saliva flow rate in older participants. The programme, however, did not result in significant changes in posterior tongue strength, cheek strength and perceived oral health., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Burning mouth syndrome (BMS) is a chronic pain condition affecting the oral cavity. This condition mostly affects peri- or postmenopausal women; for this reason, sexual hormonal changes have been implicated in BMS pathogenesis., Methods: A systematic review was performed in MEDLINE/PubMed, Scopus, Web of Science, Cochrane Library and EMBASE without restriction for language or year. Eligibility criteria were controlled studies addressing the PICO question: (P) patients with BMS; (I) detection of the sex hormones; (C) patients without BMS; (O) changes on sexual hormones as a risk factor for BMS severity. Risk of bias was performed with Newcastle-Ottawa Quality Assessment Scale., Results: Four studies were included. Salivary levels were evaluated in three studies and serum blood was used in one. Three studies analysed oestradiol and/or dehydroepiandrosterone (DHEA), two assessed progesterone and one evaluated follicle-stimulating hormone (FSH). Oestradiol results were contradictory, with two studies reporting lower levels in BMS patients compared to controls and one finding the opposite. DHEA was statistically lower in the BMS group in one study. Progesterone showed opposite results in two studies, although none with statistical significance. FSH was statistically higher in the BMS group compared to controls. Correlation of hormones with quality of life was performed in three studies and there was no significant correlation with self-perceived symptoms severity., Conclusion: Sexual hormones can be altered in BMS, especially oestradiol. Despite these changes, we did not find correlation between hormone fluctuation and BMS symptoms intensity affecting quality of life. These findings suggested the need for further investigation on hormonal alterations, which may be a promising target on BMS management., (© 2024 John Wiley & Sons Ltd.)

C-reactive protein is a systemic inflammatory biomarker that is positively associated with the development of disease. Salivary C-reactive protein (sCRP) has previously been reported to have a diurnal rhythm with higher levels upon awakening and lower levels thereafter. The aims of this study were to evaluate the stability of sCRP across two days, characterize the daily sCRP pattern, compute morning sCRP parameters, and evaluate associations with biobehavioral health in US Navy Explosive Ordnance Disposal (EOD) technicians. Seventy male EOD technicians (age = 34.9 ± 6.5 years) participated in this study, which included a tablet-based survey, measures of health and fitness, and saliva collection. In a free-living setting, participants self-collected saliva on 2 consecutive days at WAKE, WAKE+30, WAKE+60, 4p.m., and 9p.m., for a total of 10 samples. Parameters (e.g., area under the curve) were computed to characterize the morning sCRP magnitude and pattern. Pearson product-moment correlation analyses were used to assess the stability of sCRP samples and parameters across the study period and to examine associations with biobehavioral health. Average sCRP concentrations for the 2-day period were evaluated using an analysis of variance with repeated measures. The stabilities between corresponding time points on Days 1 and 2 were very high (rs = 0.87-.94, all ps ≤ 0.001). sCRP concentrations were highest at WAKE, decreased by 73.6 % at WAKE+30, and then plateaued for the rest of the day. Parameter stabilities were good to excellent (rs = 0.77-.98, all ps ≤ 0.001). We also observed associations between sCRP parameters, self-reported health behaviors, and objective measures of health and fitness. In this study of a military population, we characterized sCRP as diurnal with robust stability across 2 consecutive days, which demonstrates the feasibility of sCRP as a biomarker. These results have significant implications for study methodology and for using sCRP as a marker of dysfunction or disease., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Clozapine (CLZ) -related accidents or crimes are common in the world. Oral fluid drug detection is a convenient measure of dealing with things like that. There has not been any literature reported detailedly the representation rule of clozapine and its metabolites in oral fluid so far. The study aimed to describe the pharmacokinetics of CLZ and its metabolites N-desmethylclozapine and clozapine-N-oxide in human oral fluid after a single 12.5 mg oral dose of CLZ. Twenty-nine volunteers, including 20 males and 9 females, were recruited, and 2 mL oral fluid was collected from each participant at post-consumption time-points of prior (zero), 0.5, 1.5, 3, 5, 8, 12, 24, 36, 51, 82, and 130 h, respectively. Analytes of interest were extracted with solid-phase extraction and analyzed with liquid chromatography tandem mass spectrometry method. Pharmacokinetic parameters were calculated using the pharmacokinetic software DAS according to the non-compartment model. The maximum concentration, the time of maximum concentration, oral clearance, and the elimination half-life of clozapine were 16.57 ± 9.63 ng/mL, 4.53 ± 3.61 h, 57.65 ± 23.77 L/h and 53.58 ± 52.28 h, respectively. The maximum concentration, the time of maximum concentration, and the elimination half-life of the metabolite N-desmethylclozapine were 3.08 ± 1.19 ng/mL, 9.38 ± 9.33 h and 62.67 ± 82.57 h, respectively; of clozapine-N-oxide were 1.15 ± 0.36 ng/mL, 4.53 ± 2.19 h and 19.15 ± 23.11 h, respectively. It was the first study on the pharmacokinetics of CLZ and its metabolites in the oral fluid of Chinese healthy volunteers, and it provided a basis for the therapeutic drug monitoring and toxicological interpretation in clozapine-related cases., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A rapid, sensitive and specific test for blood is reported based on a novel application of recombinase polymerase amplification integrated with CRISPR-Cas and lateral flow assay (LFA). The blood specific marker ALAS2 was used as the target to record the presence of blood. The assay used either RNA extracted from a body fluid as a template, or omitting this extraction step and using a direct approach where the questioned body fluid was added directly to the assay. The assay only detected blood (all peripheral blood and some menstrual blood samples) and no other body fluid (semen, saliva, or vaginal fluid). The limit of detection varied from an initial template of 0.195 ng extracted RNA (2 7 dilution) or 0.0218 μL (2 6 dilution) liquid peripheral blood. The assay gave the expected result when peripheral blood was mixed with saliva: ratios of peripheral blood/saliva at 19:1, 3:1, 1:1, 1:3 and 1:19 all gave a positive result using extracted RNA. By contrast, only three ratios of peripheral blood and saliva gave a positive result for blood (19:1, 3:1 and 1:1) when adding these two body fluids directly. When peripheral blood was mixed with semen there was a strong inhibition of the assay and ALAS2 could only be detected at ratio of 19:1 using RNA. Using reconstituted peripheral bloodstains gave comparable results to liquid peripheral blood. This is the first application of RT-RPA integrated CRISPR and combined with a LFA assay to detect body fluid-specific RNA. The proposed method opens up the potential to perform this method remote from laboratories such as at crime scenes., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study explores the impact of various types of carbonation on sensory stimulation in the mouth, salivary secretion and the neurotransmitter substance P (SP), as well as body responses such as heart rate (HR) and Galvanic Skin Response (GSR). Three types of carbonation (one made using a soda machine, another carbonated with a gasifier, and the last commercial sparkling water) were used to produce different bubbles resulting in distinct sensory characteristics assessed by a trained panel. The impact of carbonation was measured by recording changes in salivary flow rate, SP levels, salivary secretory immunoglobulin A (SIgA), HR, and GSR in fifteen healthy participants. The results showed that the bubble type only affected the sensory perception of carbonation. Regardless of bubble type, carbonation increased salivary flow rate and SP values, SigA and HR. These characteristics are being sought to improve treatments for dysphagia or dry mouth. Therefore, these findings highlight the potential therapeutic application of carbonation in these situations., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Introduction: Background: stevia and D-tagatose have shown a reduction in total calorie and carbohydrate intake as a substitute for sucrose, demonstrating a stabilizing effect on pH and bacterial proliferation. Objective: to evaluate the effect of D-tagatose, stevia and sucrose on salivary pH and bacterial activity in odontology students. Methodology: a controlled study of parallel and randomized groups with a single blind, whose sample considered three groups subjected to a mouthwash of D-tagatose (n = 10), stevia (n = 10) and sucrose (n = 10). These solutions were administered over 1 minute in a single 6.4 % concentrated dose. Data collection and analysis considered the recording of salivary pH 5 min before exposure to the sweetener, immediately after expulsion of the mouthwash and 15 min later, 30 min, 45 min and 48 hours. The counting of the final number of colony-forming units per mL (CFU/mL) was counted using the salivary samples obtained immediately after exposure of the sweetener together with the sample obtained 30 minutes later, with the cultures performed on agar plates. Results: D-tagatose, stevia and sucrose presented significant differences in total CFU/mL at 30 minutes (p < 0.001), while salivary pH showed significant differences at 48 hours after administration (p < 0.001). Conclusion: D-tagatose, stevia and sucrose present significant differences in total CFU/mL and salivary pH, these findings being a possible indication of a partial inhibitory effect on bacterial metabolism.

Aim: This study aimed to investigate the effects of the two pre-ovulatory and mid-luteal phases of the menstrual cycle on cognitive function, as well as possible mediators of metabolism and salivary cortisol, at rest and after an aerobic exercise session., Study Design: Twelve active young unmarried women aged 22-30 years volunteered to participate in the study. The participants performed a 20-min exercise session on a cycle ergometer at 60-70% of their reserve heart rate twice, during the follicular (pre-ovulation: days 7-10) and luteal (mid-luteal: days 21-24) phases of the menstrual cycle. Saliva samples were collected to measure cortisol. Fat utilization, respiratory exchange ratio (RER), and energy expenditure (during exercise) were measured using a spiroergometer. Cognitive function was assessed using the Stroop test. Cognitive function and cortisol levels were measured before and after each exercise session., Results: The findings of this study indicated no significant differences in variables during the resting follicular and luteal phases. Cortisol levels and cognitive function were increased after exercise compared with before exercise in both the follicular and luteal phases. Cortisol and fat utilization after exercise were significantly higher in the follicular phase than in the luteal phase. There were no significant differences between the follicular and luteal phasesregarding the effects of exercise on cognitive function, energy expenditure, and RER., Conclusion: In general, the follicular and luteal phases of menstruation may not affect cognitive function in response to a single aerobic exercise session, although they change some metabolic factors and cortisol., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Mirzaei Khalil Abadi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

A simple and rapid qualitative chromatographic method with a unique extraction approach was developed and validated to screen oral fluid samples for 31 compounds in driving under the influence of drugs investigations. The scope and sensitivity of the method meets or exceeds Tier I recommendations established by the National Safety Council's Alcohol, Drugs and Impairment Division. Since this is a targeted chromatographic screen (rather than an immunoassay), cutoffs were set to match the confirmation levels in the recommendations. Sample preparation involved a single-step liquid-liquid extraction procedure, using a mixture of methyl tert-butyl ether, isopropanol, and hexane and was applied to samples collected with the Quantisal™ device. Instrument analysis was conducted by liquid chromatography-tandem mass spectrometry, using a Restek Raptor™ biphenyl column for chromatographic separations and a total run time of 8 min. Validation results met all requirements of ANSI/ASB Standard 036 (1st edition)-Standard Practices for Method Validation in Forensic Toxicology., (© The Author(s) 2024. Published by Oxford University Press. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site–for further information please contact journals.permissions@oup.com.)

Background: The frequent occurrence of dental caries and periodontal diseases in women during pregnancy may be due to many factors, such as salivary variables. The aim of this study was to evaluate the effects of pregnancy on salivary pH, flow rate, the DMFT index, and CPI sores., Methods: A total of 198 volunteers (pregnant in different trimesters and non- pregnant) were included for the present observational cross-sectional study. Data about sociodemographic characteristics and dental and systemic health conditions were recorded. Unstimulated saliva samples were collected for 5 min via the spitting method. The pH of the saliva was measured by a portable pH meter. The salivary flow rate was determined by the weight measurement method. The DMFT index and CPI were determined. The statistical evaluation was performed using Robust ve Poisson Regression analyses (p < 0.05)., Results: It was determined that there was a gradually significant decrease in the Ph value from the first trimester to the third trimester during pregnancy, which was lower than the control group. (p < 0.001) The flow rate of pregnants in their third trimester was lower than that of first trimester (p = 0.017). The CPI scores of pregnant women were significantly greater than that of non-pregnants (p = 0.042), while the DMFTs were similar among all groups., Conclusion: Our findings indicate that pregnancy leads to a notable reduction in unstimulated salivary pH and flow rate, which also has a detrimental impact on periodontal health., Trial Registration: Clinical Trials-ID: NCT06343337; Registration Date: 04.01.2024., (© 2024. The Author(s).)

Background and Purpose: Despite substantial evidence supporting the role of resident bacterial communities in therapeutic fasting outcomes, research has primarily focused on gut microbiota, leaving changes in oral microbiota largely unexplored. The clinical significance of oral health changes during fasting is nonetheless underscored by the documented development of halitosis in fasting individuals. However, no scientific studies have comprehensively examined the interplay between salivary microbiota alterations, inflammatory changes in the gingival crevice, and the production of malodorous volatile compounds. We examined volatile sulphur compounds (VSC) in breath during fasting, cytokine levels in the gingival crevice, and oral microbiota composition of the saliva in a single-arm interventional study involving 36 subjects who fasted for 10 ± 3 days., Materials and Methods: Participants fasted according to Buchinger fasting guidelines. VSC were evaluated every morning before any food or drink intake using the OralChroma gas chromatography device. Saliva and gingival crevicular fluid (GCF) samples were collected at the clinical site before fasting, at the end of fasting, and at the end of food reintroduction. Follow-up saliva samples were sent to the patients after 1 and 3 months. Saliva samples were processed and analysed by targeted sequencing of 16S rRNA gene amplicons, whereas the expression of 6 inflammatory markers in the GCF were analysed using a multiplex fluorescent bead-based immunoassay., Results: The quantification of volatile compounds in the breath demonstrated a statistically significant increase in dimethylsulfide levels during fasting, which corroborates the occurrence of bad breath as a common side effect of fasting. Salivary microbiota profiling showed a shift in microbial composition, including reduction in the levels of Neisseria, Gemella and Porphyromonas spp., concomitant with an increase in the levels of Megasphaera, Dialister, Prevotella, Veillonella, Bifidobacteria, Leptotrichia, Selenomonas, Alloprevotella, and Atopobium. We further demonstrated a reduction in the levels of the pro-inflammatory cytokine interleukin-8 in the GCF., Conclusion: Dimethylsulfide concentrations in the breath increased during fasting, and this was correlated to changes in the oral microbiota. Future studies are needed to illuminate the possible impact of these changes on oral and general health status.

Seeding activities of disease-associated α-synuclein aggregates (αSyn D ), a hallmark of Parkinson's disease (PD), are detectable by seed amplification assay (αSyn-SAA) and being developed as a diagnostic biomarker for PD. Sensitive and accurate αSyn-SAA for blood or saliva would greatly facilitate PD diagnosis. This prospective diagnostic study conducted αSyn-SAA analyses on serum and saliva samples collected from patients clinically diagnosed with PD or healthy controls (HC). 124 subjects (82 PD, 42 HC) donated blood and had extensive clinical assessments, of whom 74 subjects (48 PD, 26 HC) also donated saliva at the same visits. An additional 57 subjects (35 PD, 22 HC) donated saliva and had more limited clinical assessments. The mean ages were 69.21, 66.55, 69.58, and 64.71 years for PD serum donors, HC serum donors, PD saliva donors, and HC saliva donors, respectively. αSyn D seeding activities in either sample type alone or both sample types together were evaluated for PD diagnosis. Serum αSyn-SAA data from 124 subjects showed 80.49% sensitivity, 90.48% specificity, and 0.9006 accuracy (AUC of ROC); saliva αSyn-SAA data from 131 subjects attained 74.70% sensitivity, 97.92% specificity, and 0.8966 accuracy. Remarkably, the combined serum and saliva αSyn-SAA from 74 subjects with both sample types achieved better diagnostic performance: 95.83% sensitivity, 96.15% specificity, and 0.98 accuracy. In addition, serum αSyn D seeding activities correlated inversely with Montreal Cognitive Assessment in males and positively with Hamilton Depression Rating Scale in females and in the < 70 age group, whereas saliva αSyn D seeding activities correlated inversely with age at diagnosis in males and in the < 70 age group. Our data indicate that serum and saliva αSyn-SAA together can achieve high diagnostic accuracy for PD comparable to that of CSF αSyn-SAA, suggesting their potential utility for highly sensitive, accurate, and minimally invasive diagnosis of PD in routine clinical practice and clinical studies., (© 2024. The Author(s).)

Introduction: Exposure to secondhand smoke (SHS) risks children's health. However, biomarkers are rarely used to study SHS exposure among children in low- and middle-income countries., Aims and Methods: We analyzed cross-sectional data collected between March and November 2022 for a cluster-randomized controlled trial investigating a Smoke-Free Intervention in 2769 children aged 9-15 in 74 schools (34 in Dhaka, Bangladesh, and 40 in Karachi, Pakistan). Children's saliva was tested for the concentration of cotinine-a highly sensitive and specific biomarker for SHS exposure. Based on their reports, children's homes were categorized as Nonsmoking Homes (NSH) when residents were nonsmokers; Smoke-free Homes (SFH) when residents and visitors smoked outdoors only; and Smoke-permitted Homes (SPH) when either residents or visitors smoked indoors. We compared cotinine concentrations across these home types and the two cities using a proportional odds model., Results: Overall, 95.7% of children (92% in Dhaka; and 99.4% in Karachi) had cotinine levels between 0.1 and 12 ng/mL, indicating SHS exposure. Median cotinine levels were higher in Karachi (0.58 ng/mL, IQR 0.37 to 0.93) than in Dhaka (0.27 ng/mL, IQR 0.16 to 0.49). Median cotinine concentration was also higher among children living in SPH than those in either NSH or SFH; with absolute differences of approximately 0.1-0.3 and 0.05 ng/mL, respectively., Conclusions: The level of SHS exposure in Dhaka and Karachi indicates widespread and unrestricted smoking. Smoking restrictions in households and enforcement of smoking bans are urgently needed., Implications: The high levels of SHS exposure in children living in SFH suggest parental behavior to hide their smoking and/or exposure in private vehicles or public spaces. It is important to advocate for SFH and cars to protect children from SHS exposure. However, these initiatives alone may not be enough. There is a need to enforce smoking bans in enclosed public places and transportation, as well as extend these bans to playgrounds, parks, fairgrounds, and other public spaces that children frequently visit. It is essential to complement smoking restrictions with tobacco cessation advice and support in these settings., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Research on Nicotine and Tobacco.)

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.

Background: Poor physical fitness, stress, and fatigue are factors impacting military readiness, national security, and economic burden for the United States Department of Defense. Improved accuracy of wearable biosensors and remote field biologic sample collection strategies could make critical contributions to understanding how physical readiness and occupational stressors result in on-the-job and environment-related injury, sleep impairments, diagnosis of mental health disorders, and reductions in performance in war-fighters., Objective: This study aimed to evaluate the feasibility and acceptability of intensive biomarker and biometric data collection to understand physiological and psychological stress in Army Reserved Officer Training Corps cadets before, during, and after a 96-hour field training exercise (FTX)., Methods: A prospective pilot study evaluated the feasibility and acceptability of multimodal field data collection using passive drool saliva sampling, sweat sensors, accelerometry, actigraphy, and photoplethysmography. In addition, physical fitness (Army Combat Fitness Test), self-reported injury, and psychological resilience (Brief Resilience Scale) were measured., Results: A total of 22 cadets were included. Two were lost to follow-up due to injury during FTX, for a retention rate of 91%. Assessments of performance and psychological resilience were completed for all remaining participants, resulting in 100% testing adherence. All participants provided saliva samples before the FTX, with 98% adherence at the second time point and 91% at the third. For sweat, data collection was not possible. Average daily wear time for photoplethysmography devices was good to excellent, meeting a 70% threshold with data collected for ≥80% of person-days at all time points. Of the participants who completed the FTX and 12 completed a post-FTX acceptability survey for a response rate of 60%. Overall, participant acceptance was high (≥80%) for all metrics and devices., Conclusions: This study demonstrates that wearable biosensors and remote field biologic sample collection strategies during a military FTX have the potential to be used in higher stakes tactical environments in the future for some, but not all, of the strategies. Overall, real-time biometric and biomarker sampling is feasible and acceptable during field-based training and provides insights and strategies for future interventions on military cadet and active-duty readiness, environmental stress, and recovery., (©Rachele Pojednic, Amy Welch, Margaret Thornton, Meghan Garvey, Tara Grogan, Walter Roberts, Garrett Ash. Originally published in JMIR Formative Research (https://formative.jmir.org), 18.10.2024.)

Background: It is well-recognized that maintaining a regular yogic practice is associated with numerous physical and psychological health benefits. However, few studies have explored the possible psychological and neurophysiological mediators through which the component parts of yoga-ethics, breath regulation, postures, and meditation-work to produce salutary effects., Purpose: To address this gap, we conducted a cluster randomized trial to test the following set of theory-based mechanisms: emotion regulation for ethics, self-control for breath regulation, discomfort and distress tolerance for postures, and mindfulness for meditation. We also explored yoga's effects on the autonomic nervous system by examining salivary acetylcholinesterase levels., Methods: Participants (N = 260) were randomly assigned in clusters (n = 37) to a single, hour-long standard or postures-only yoga class., Results: Findings suggest that a single yoga class may confer both psychological and neurophysiological benefits, yet there were few differences between the two types of yoga classes. Pre- to post-session main effects of time, all in the expected direction, emerged for five of eight theoretical mediators, as well as for salivary acetylcholinesterase levels. Time X condition interactions observed for three of the mediators-cognitive reappraisal, discomfort tolerance, and expressive suppression-along with findings from the exploratory mediation analysis suggest potential unique benefits of the two yoga sessions for certain outcomes., Conclusions: Additional studies are needed to replicate these results and to test other potential mediators and/or primary outcomes through which yoga might work to promote health., (© Society of Behavioral Medicine 2024. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The aim of the present study was to investigate whether the wearing of hearing protection has a positive influence on stress levels during an MRI examination in dogs under anaesthesia. To this end, the stress hormones cortisol and arginine vasopressin (AVP) were measured in the saliva of patients wearing hearing protection during an MRI scan, as well as in the control group without hearing protection, before and after the scan. Pulse rate and noise level were also measured during the MRI. It was shown that salivary cortisol concentrations in dogs without hearing protection increased during the MRI examination, while those in the control group with hearing protection remained the same (p<0.05). The pulse range was greater in the control group without hearing protection, although not statistically significant. The maximum loudness during the MRI examination, measured at 200 cm from the gantry, was 87 dB(A). The results on the loudness during mri scan highlight that hearing protection should always be used to minimise hearing damage and possibly the stress level for dogs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Schroers et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Successful reproductive management of domestic mammals depends primarily upon timely identification of oestrous cycle stages. There is a need to develop an alternative non-invasive, welfare-friendly, accurate and reliable method to identify reproductive cycle stages. This is of particular interest for horse breeders, because horses are high-value farm animals that require careful management and individual monitoring. Saliva sampling is non-invasive, painless and welfare-friendly. Thus, we performed a metabolomic analysis of equine saliva during different reproductive stages to identify changes in the salivary metabolome during anoestrus, the oestrous cycle and early gestation. We compared the saliva and plasma metabolomes to investigate the relationship between the two fluids according to the physiological stage. We collected saliva and plasma samples from six mares during seasonal anoestrus, during the follicular phase 3 days, 2 days and 1 day before ovulation and the day when ovulation was detected, during the luteal phase 6 days after ovulation, and during early gestation 18 days after ovulation and insemination. Metabolome analysis was performed by proton-nuclear magnetic resonance spectroscopy. We identified 58 and 51 metabolites in saliva and plasma, respectively. The levels of four metabolites or groups of metabolites in saliva and five metabolites or groups of metabolites in plasma showed significant modifications during the 4 days until ovulation, ie 3 days prior to and on the day of ovulation. The levels of 11 metabolites or groups of metabolites in saliva and 17 metabolites or groups of metabolites in plasma were significantly different between the seasonal anoestrus and the ovarian cyclicity period. The physiological mechanisms involved in the onset of ovarian cyclicity and in ovulation induced modifications of the metabolome both in plasma and saliva. The metabolites whose salivary levels changed during the reproductive cycle could be potential salivary biomarkers to detect the reproductive stage in a welfare friendly production system. In particular, we propose creatine and alanine as candidate salivary biomarkers of ovulation and of the onset of ovarian cyclicity, respectively. However, extensive validation of their reliability is required. Our study contributes to extend to domestic mammals the use of saliva as a non-invasive alternative diagnostic fluid for reproduction in a welfare-friendly production system., Competing Interests: Declaration of competing interest None., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Thermal comfort in an office impacts physical health, stress, and productivity. Humidity affects thermal comfort; however, the underlying mechanism remains unclear. This study assessed the influence of humidity on body temperature, thermal comfort, stress, and their relationship in working individuals. Thirteen participants performed three sets of 20-min calculation tasks followed by a 10-min rest in 26 °C or 33 °C with relative humidity (RH) of 30 % or 60 %. Core body temperature (T core ), mean skin surface temperature (T skin ), and electrocardiogram were continuously recorded. Subjective thermal sensations and comfort were assessed with visual analog scales. Stress level was estimated based on α-amylase activity and immunoglobulin A level in saliva and heart rate variability. Mean T skin and T core elevated at 33 °C with 60 % RH, where warm sensation and thermal discomfort also increased. Heart rate variability reflecting parasympathetic nerve activity decreased. There was a negative linear relationship between weighted body temperature and thermal comfort. However, thermal discomfort was augmented at a given weighted body temperature at 60 % RH. Thus, under indoor working conditions, high humidity may augment thermal discomfort and become a stress factor. Increases in T skin and T core are involved in the mechanism, alongside other factors., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this study., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

The stereochemical stability of the popular drugs of abuse 2-, 3- and 4-chloromethcathinone was studied in the mobile phase used for the isolation of their enantiomers by high-performance liquid chromatography, as well as in various biological matrixes such as whole blood, saliva and urine. For 2-, 3-, and 4-chloromethcathinones the rate constants and half-lives of their first order racemization reaction was assessed. It was found that at 25 °C the racemization rate constant decreases in the order 2-CMC > 3-CMC > 4-CMC while their stereochemical stability in biological matrixes decreases in the order urine > saliva > whole blood. This information must be considered for the adequate storage of purified enantiomers in the collected fractions, as well as in the studies focused on their enantioselective transformation in the human body., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: 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., Methods: 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., Results: 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., Conclusions: 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., Competing Interests: Manuel Jimenez is an Academic Editor for PeerJ., (©2024 Ferreira et al.)

Background: The aim of this study was to investigate the microbiome shift of denture biofilm formation, the incidence of Candida and changes of salivary inflammatory markers following insertion of 3-dimensional printed PMMA denture., Methods: This was a pilot study of 6 patients provided with 3D printed (test group) or conventionally heat-cured (control group) removable partial acrylic dentures followed up for 6 weeks. 3 denture swabs were collected at week 1, 3 and 6; and saliva samples were collected at baseline, week 1, 3 and 6 following denture insertion. Microbial DNA was isolated, and the 16S rRNA gene was amplified and sequenced to assess the denture microbiota. The presence of Candida was determined using PCR assay. The levels of salivary biomarkers lactoferrin (LTF) and histatin (HTN) were determined using ELISA., Results: Denture microbiome of both groups exhibited similar microbial compositions at weeks 1, 3 and 6. However, the percentage of Streptococcus in the test group was significantly lower at week 1 (p < 0.05). Greater bacterial diversity was detected in the test group (p < 0.05; pairwise Wilcoxon) after 6 weeks. The presence of Candida was only detected in one sample of the 3D printed denture (test group) after 6 weeks. The level of LTF biomarkers increased in both groups after 6 weeks, however, the level of HTN increased only in the control group., Conclusions: Denture microbiome of both groups demonstrated similar microbial compositions. After 6 weeks, 3D printed denture demonstrated higher diversity with delayed microbiome shift compared to conventional heat cure dentures. 3D printed denture may be a viable alternative to conventional denture and it has a potential to delay microbial dysbiosis., Clinical Relevance: Among wearers, 3D printed denture can potentially reduce the risk of denture stomatitis. A greater diversity may delay or slow the transition of denture microbiome to dysbiosis., (© 2024. The Author(s).)

Objectives: 8-hydroxydeoxyguanosine (8-OHdG) and Malondialdehyde (MDA) are commonly used as markers to evaluate oxidative DNA and Lipid damage in disorders including chronic inflammatory diseases. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) protect tissues against oxidative injury from free oxygen radicals generated by various metabolic processes. The aim of this study was to evaluate 8-OHdG and MDA levels, and SOD and GPx activities in whole saliva of patients with peri-implant diseases., Materials and Methods: A cross-sectional study was conducted on a sum of 60 age gender balanced; peri-implantitis (n = 20), peri-mucositis (n = 20) and healthy (n = 20) individuals. Unstimulated whole saliva samples were collected and to determine the clinical condition of each subject; the plaque index (PI), gingival index (GI), peri-implant probing pocket depth (PIPD), peri-implant presence of bleeding on probing (BOP) (with/without suppuration) and radiographic signs of crestal bone loss (BL) were measured. The salivary 8-OHdG level was measured using the ELISA method. SOD, GPx activities and MDA levels were determined spectrophotometrically., Results: A total of 60 individuals had evaluations of 318 implants. In comparison to the peri-mucositis and peri-implantitis groups, the healthy group had significantly lower PI and GI scores (p < 0.001). The PIPD value differed amongst the groups, with the peri-implantitis group having the highest value (p < 0.001). Compared to the peri-mucositis and control groups, the peri-implantitis group had a significantly higher BL score (p < 0.001 and p < 0.001, respectively). The peri-implantitis group showed a significantly higher 8-OHdG level (p < 0.001; p < 0.001 respectively) than the peri-mucositis and control groups. Compared to the peri-mucositis and control groups, the peri-implantitis group had a significantly higher MDA level (p < 0.001 and p < 0.001, respectively). The peri-implantitis group had a significantly higher SOD level (p < 0.001 and p < 0.001, respectively) in comparison to the peri-mucositis and control groups. There was no significant difference in GPx levels between the peri-mucositis and control groups (p > 0.05), while the peri-implantitis group had significantly lower GPx levels than the peri-mucositis and control groups (p < 0.001 and p < 0.001, respectively)., Conclusions: Elevated levels of oxidative stress in saliva may indicate the onset of pathological bone loss surrounding the implant and may be an indication of peri-implantitis., Clinical Relevance: In peri-implant diseases, changes may occur in the levels of 8-OHdG, MDA, SOD and GPx in saliva, which may lead to a deterioration in the oxidant/antioxidant balance., (© 2024. The Author(s).)

Background: Enhancing the prognosis and treatment outcomes of oral cancer relies heavily on its early detection. This study aims to establish a connection between serum and salivary dipeptidyl peptidase-4 (DPP-4) levels and oral squamous cell carcinoma (OSCC), comparing them with oral potentially malignant lesions (OPMLs) and control subjects and validating salivary DPP-4 as a diagnostic marker for the early detection of oral cancer., Methodology: Forty-five systemically healthy individuals were categorized into three groups: Group I consisted of 15 patients diagnosed with OSCC, Group II comprised 15 patients with OPMLs (including leukoplakia and oral lichen planus), and Group III included 15 participants without any oral mucosal lesions. Serum and whole unstimulated salivary samples were collected from all participants to assess DPP-4 levels using an enzyme-linked immunosorbent assay (ELISA) kit. Receiver operating characteristic (ROC) analysis was conducted to determine the area under the curve (AUC), sensitivity, specificity, and diagnostic accuracy of DPP-4., Results: Both serum and salivary DPP-4 levels were highest in the healthy group, followed by OPMLs, and lowest in the OSCC group, with statistically significant differences observed. ROC analysis demonstrated excellent diagnostic accuracy of salivary DPP-4 in distinguishing OSCC from healthy individuals, OPMLs from healthy individuals, and OSCC from OPMLs, with accuracies of 100%, 100%, and 96.67%, respectively. Salivary DPP-4 levels also exhibited a statistically significant correlation with OSCC grades., Conclusions: DPP-4 appears to play a protective, anti-oncogenic role in maintaining oral tissue health. The remarkable diagnostic accuracy of both serum and salivary DPP-4 in discriminating between OSCC, OPMLs, and healthy controls suggests its potential utility as a well-established marker for early oral cancer diagnosis. Salivary DPP-4, being non-invasive, could serve as a convenient chair-side diagnostic tool for the early detection of OSCC., Clinical Trial Registration: The study was retrospectively registered on clinicaltrial.gov with NCT06087042, date of registration (first posted date): 17/10/2023., (© 2024. The Author(s).)

Background: Individuals who provide critical emergency care mount rapid psychobiological responses when faced with an incident. These responses are adaptive and ensure resources at time of demand; however, frequent activation with minimal opportunity for recovery can have negative consequences for health and wellbeing. Monitoring individuals in real emergency situations would provide an understanding of their stress responses during the provision of critical care; however, this presents logistical challenges. An alternative is to assess individuals during high-fidelity training scenarios. This is the first comprehensive assessment of psychobiological responding during continuous high-fidelity training in pre-hospital emergency medicine., Methods: A sample of doctors and paramedics (N = 27) participated during 10 days of training and a weekend of no activities. Training involved the acquisition of human factors, non-technical and surgical skills, and their application in complex high-fidelity scenarios including road-traffic accidents, firearms incidents, and swift water rescue operations. On each day participants reported levels of state, cognitive, and somatic anxiety, and self-confidence following waking and before sleep, and their anticipated (at wake) and experienced (before sleep) demands of the day. Saliva samples were obtained each day for assessment of diurnal cortisol indices and the Cortisol Awakening Response (CAR). Garmin smartwatches were worn throughout for the collection of heart rate and HRV-derived stress., Results: There were significant (p < 0.001) differences across days for state, cognitive, and somatic anxiety; self-confidence; anticipated and experienced demands; aggregated measures of heart rate and HRV-derived stress; levels of cortisol at waking (p = 0.002) and for the CAR (p < 0.001). Measures of psychobiological responding during training were distinct from the weekend and the highest levels of psychobiological responding occurred on days characterised by greater anticipated and experienced demands., Discussion: This high-fidelity training is typical of the day-to-day requirements of emergency services and these observations are representative of functioning during real-life critical care emergencies. Increased responding during times of demand is adaptive; however, frequent and sustained responding increases allostatic load and is a contributor to burnout. As burnout is a significant concern in emergency medicine, this study identifies patterns of responding and recovery that may impact upon longer-term health and wellbeing., (© 2024. The Author(s).)

Emerging technology in the new era of sensors to detect and quantify neurological reaction-based research has demanded the development of sensors for the neurotransmitter dopamine (DA). In recent decades, electrochemical sensors have offered rapid and sensitive detection of DA, but the presence of interfering compounds, such as uric acid (UA) and ascorbic acid (AA), poses a great threat to the development of DA sensors. Additionally, reusing traditional methods leads to challenges like prolonged preparation and expensive instruments. This research work offers a nanohybrid two-dimensional (2D) paper-like graphene oxide (GO) and three-dimensional (3D) cerium oxide nanosphere (CeONS) heterostructure composite (G-CeONS) created via stoichiometric synthesis for the non-enzymatic detection of DA oxidation in the presence of other complex biological compounds. The constructed G-CeONS nanohybrid composite enables enhanced selectivity and sensitivity towards DA detection through its interfacial engineering. The heterostructure formation of a 2D nanosheet draped over 3D nanospheres exhibits a wide linear concentration range of 100-30 800 nM with a low detection limit of 20.98 nM. Further investigation of the real-time performance on human saliva and DA injections afforded prominent results. In addition, the synergetic effect of G-CeONS improves DA detection accuracy and reliability towards enabling transformational neurochemical and medicinal applications.

A Raman-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, H 2 O 2 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 882 cm -1 . Meanwhile, the original characteristic peak at 998 cm -1 remained unchanged. Therefore, the I 882 /I 998 ratio increased with increasing content of D-AAs in the sample to be tested, allowing D-AAs to be quantitatively detected. The Au/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 10.1 µM and 13.7 µM throughout the linear range of 20-500 µM. The good results of the saliva detection suggested that this SERS sensor could rapidly differentiate between early-stage gastric cancer patients and healthy individuals., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Objectives: The purpose of this review was to present the basic concepts of metabolomics methodology and the use of saliva for diagnostic, prognostic, and predictive strategies., Material and Methods: This review followed the focus in: "saliva metabolomics" and "oral diseases". The authors searched studies on PubMed database. The inclusion criteria were original studies and reviews that assessed metabolomics techniques. A descriptive analysis was performed considering the study design, approach system, clinical steps, and tools for the determination of profile or biomarkers metabolites, and the advantages and disadvantages., Results: Metabolomic analyses use a combination of analytical instrumentation and informatic tools to provide information on metabolite characteristics. In this review we described different technologies applied and the advantages and limitations of each technique. Furthermore, in the literature search, we retrieved 25 studies that investigated saliva metabolites in oral diseases: 8 studies used targeted analysis and 17 untargeted metabolomics approaches. Most studies included patients with periodontal diseases, oral squamous cell carcinoma, and Sjögren Syndrome. The most frequently reported metabolites were glycine, leucine, phenylalanine, dipeptides, linoleic acid, arachidonic acid, tyrosine, choline, taurine, lactate, valine, and proline., Conclusions: Metabolomics analysis has emerged as a powerful tool for tumor diagnosis and to enhance tumor classification, including salivary gland tumors (SGTs). It also holds promise for developing personalized treatment plans and defining more precise prognostic categories., Clinical Relevance: Metabolomics is the most functional and comprehensive technique for monitoring and understanding gene functions and identifying the biochemical state of an organism in response to genetic and environmental changes., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: To investigate the preventive and discoloring effects of a single and two weekly applications of 38% silver diamine fluoride (SDF) against dentin erosion., Materials and Methods: 180 dentin blocks were divided into four groups. Group 1 (SDF2) received two weekly applications of 38% SDF. Group 2 (SDF1) received a single application of 38% SDF. Group 3 (SNF) received a daily application of stannous chloride/amine fluoride/sodium fluoride (standard of care for dental erosion). Group 4 (DW) received a daily application of deionized water. The treated blocks were subjected to a 14-day erosive challenge. Crystal characteristics, elemental composition, surface morphology, percentage of surface microhardness loss (%SMHL), surface loss, and color change (ΔE) were investigated using X-ray diffraction (XRD), energy-dispersive spectrometry (EDS), scanning electron microscopy (SEM), hardness testing, profilometry, and digital spectrophotometry, respectively., Results: XRD and EDS showed dentin surfaces had silver compounds in SDF2 and SDF1, and stannous chloride in SNF. SEM revealed less dentin demineralization with tubular occlusion in SDF2, SDF1, and SNF, but severe demineralization in DW. The %SMHL of SDF2, SDF1, SNF, and DW were 10.8 ± 2.1, 15.7 ± 2.1, 17.9 ± 2.1, and 28.7 ± 2.0 (SDF2 < SDF1 < SNF < DW, p < 0.05). Surface loss (µm) of SDF2, SDF1, SNF, and DW were 5.0 ± 0.6, 6.0 ± 0.6, 6.0 ± 0.7, and 9.0 ± 0.5 (SDF2 < SDF1 = SNF < DW, p < 0.001). ΔE of SDF2, SDF1, SNF, and DW were 26.0 ± 3.4, 12.1 ± 3.8, 6.9 ± 3.5, and 3.9 ± 3.6 (SDF2 > SDF1 > SNF = DW, p < 0.001)., Conclusion: 38% SDF with two weekly applications provided better preventive effects against dentin erosion, but it might discolor dentin., Clinical Relevance: The increased 38% SDF application showed a better anti-erosive potential against dentin erosion. However, SDF caused black staining on the dentin., (© 2024. The Author(s).)