Salivary immunoglobulin levels and periodontal indices in Brazilian children with and without type 1 diabetes.

This cross-sectional study evaluated the association between salivary immunoglobulins, plaque index, and gingival index in Brazilian children with and without type 1 diabetes mellitus (DM1). The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklist for the reporting of observational studies was followed. The DM1 group had 38 children, and an equal number of volunteers matched by sex and age were recruited as controls. Clinical examination was performed for plaque index and gingival index determination. Non-stimulated whole saliva was collected. Concentrations of IgA, IgG, and IgM were determined by ELISA test. Data were tested by the Kolmogorov-Smirnov, Mann-Whitney, and Spearman tests and a multiple linear regression model (p<0.05) was performed. Gingival index was higher in the Control (DM1: 0.16±0.17; Control: 0.24±0.23, p=0.040). In DM1, there was a correlation between IgA and age (rho=0.371, p=0.024), IgM and IgG (rho=0.459, p=0.007), and IgM and gingival index (rho=0.394, p=0.014). In DM1, multiple linear regression showed that age (p=0.041; ß=0.363), gingival index (p=0.041; ß=0.398), and plaque index (p=0.008; ß=-0.506) were good predictors of IgA levels in saliva. Thus, IgA was the only researched immunoglobulin that was directly associated with plaque and gingival indices in Brazilian children with DM1, but not in control subjects.

[The effect of vitamin D deficiency on the content of some soluble signaling molecules in the oral fluid in individuals with dental caries]. / Vliyanie nedostatka vitamina D v organizme na soderzhanie nekotorykh rastvorimykh signal'nykh molekul v rotovoi zhidkosti pri kariese zubov.

OBJECTIVE: The aim of the sthudy is to sthudy the level of soluble Immune Checkpoint Molecules (B7.2, CTLA-4, Tim-3, Lag-3, PD-1) in the oral fluid during dental caries with the background of a lack and/or deficiency of 25-hydroxy-vitamin D in body. MATERIALS AND METHODS: During the research 3 groups of people were formed, each one of them included 17 people aged from 20 to 24 years. The first group included students with high-intensity caries (above 9 DMFt index) and 25-hydroxy-vitamin D levels in blood serum >30 ng/ml, the second included students with high caries intensity and 25-hydroxy-vitamin D levels <30 ng/ml. The control group consisted of students with an average DMFt index of 1.5 (from 0 to 3) and a level of 25(OH)D in the blood more than 30 ng/ml. To determine the content of B7.2 (CD86), CTLA-4, Tim-3, Lag-3, PD-1, the Human Vascular Inflammation Panel 1 multiplex analysis kit from Biolegend (USA) was used. RESULTS: The results of the research showed that during dental caries with a normal level of 25-hydroxy-vitamin D there are no significant changes in the content of Immune Checkpoint Molecules. With the background of deficiency and lack of 25-hydroxy-vitamin D there is a decrease in the amount of B7.2, LAG-3, Tim-3 and PD-1. These changes are being aggravated with an increase of the caries intensity. CONCLUSION: Vitamin D deficiency leads to a decrease in mucosal immunity of the oral cavity, the multiplication of pathogenic microorganisms, which in turn, releasing various metabolites, including cytokine-like substances, aggravate the pathological process and intensify carious lesions.

Cold pressor stress effects on cardiac repolarization.

The cold pressor test (CPT) elicits strong cardiovascular reactions via activation of the sympathetic nervous system (SNS), yielding subsequent increases in heart rate (HR) and blood pressure (BP). However, little is known on how exposure to the CPT affects cardiac ventricular repolarization. Twenty-eight healthy males underwent both a bilateral feet CPT and a warm water (WW) control condition on two separate days, one week apart. During pre-stress baseline and stress induction cardiovascular signals (ECG lead II, Finometer BP) were monitored continuously. Salivary cortisol and subjective stress ratings were assessed intermittently. Corrected QT (QTc) interval length and T-wave amplitude (TWA) were assessed for each heartbeat and subsequently aggregated individually over baseline and stress phases, respectively. CPT increases QTc interval length and elevates the TWA. Stress-induced changes in cardiac repolarization are only in part and weakly correlated with cardiovascular and cortisol stress-reactivity. Besides its already well-established effects on cardiovascular, endocrine, and subjective responses, CPT also impacts on cardiac repolarization by elongation of QTc interval length and elevation of TWA. CPT effects on cardiac repolarization share little variance with the other indices of stress reactivity, suggesting a potentially incremental value of this parameter for understanding psychobiological adaptation to acute CPT stress.

Exogenous melatonin's effect on salivary cortisol and amylase: A randomized controlled trial.

This study aimed to examine the effect of acute exogenous melatonin administration on salivary cortisol and alpha-amylase (sCort and sAA) as representatives of the HPA axis and the sympathetic nervous system, respectively. A single-dose prolonged-release melatonin (2 mg) or a placebo tablet was given to healthy volunteers (n = 64) at 20:00 h in a crossover design. The saliva was collected at six time points (20:00, 21:00, awakening, 30 min after awakening, 10:00, and 12:00 h) and was measured for sCort, sAA, and salivary melatonin (sMT) levels. Pulse rates and sleep parameters were also collected. Melatonin was effective in improving sleep onset latency by 7:04 min (p = .037) and increasing total sleep time by 24 min (p = .006). Participants with poor baseline sleep quality responded more strongly to melatonin than participants with normal baseline sleep quality as they reported more satisfaction in having adequate sleep (p = .017). Melatonin administration resulted in higher sCort levels at awakening time point (p = .023) and a tendency of lower sAA levels but these were not significant. Melatonin ingestion at 20:00 h resulted in a marked increase in sMT levels at 21:00 h and remained higher than baseline up to at least 10:00 h (p < .001). Melatonin increases sCort levels at certain time point with a tendency to lower sAA levels. These opposing effects of melatonin suggested a complex interplay between melatonin and these biomarkers. Also, the results confirmed the positive acute effect of a single-dose melatonin on sleep quality.

High-throughput proteomic analysis of extracellular vesicles from saliva by chemical probe-based array.

Extracellular vesicles (EVs) are cell-released, nucleus-free particles with a double-membrane structure that effectively prevents degradation of internal components by a variety of salivary enzymes. Saliva is an easily accessible biofluid that contains a wealth of valuable information for disease diagnosis and monitoring and especially reflect respiratory and digestive tract diseases. However, the lack of efficient and high-throughput methods for proteomic analysis of salivary biomarkers poses a significant challenge. Herein, we designed a salivary EV amphiphile-dendrimer supramolecular probe (SEASP) array which enables efficient enrichment and in situ detection of EVs protein biomarkers. Detergent Tween-20 washing of SEASP arrays removes high abundance of heteroproteins from saliva well. This array shows good analytical performance in the linear range of 10 µL-150 µL (LOD = 0.4 µg protein, or 10 µL saliva), exhibiting a good recovery (80.0 %). Compared to ultracentrifugation (UC), this procedure provides simple and convenient access to high-purity EVs (1.3 × 109 particles per mg protein) with good physiological status and structure. Coupling with mass spectrometry based proteomic analysis, differentially expressed proteins as selected asthma biomarkers have been screened. Then, we validated the proteomics primary screening results through clinical samples (100 µL each) using the SEASP array. Utilizing the dual antibody fluorescence technology, SEASP enables the simultaneous high-throughput detection of two proteins. Therefore, the EVs marker protein CD81 could be used as an internal standard to normalize the number of EVs, which was stably expressed in EVs. Proteomics and array results suggested that HNRNPU (P = 4.9 * 10-6) and MUC5B (P = 4.7 * 10-11) are promising protein biomarkers for infantile asthma. HNRNPU and MUC5B may be associated with disease onset and subtypes. The SEASP arrays provide a significant advancement in the field of salivary biomarker. The array enables high-throughput in situ protein detection for highly viscous and complex biological samples. It provides a rapid, low-cost, highly specific screening procedure and experimental basis for early disease screening and diagnosis in the field of liquid biopsy.

Interplay of hormones and metabolite excretion with fern pattern prove saliva as a potent indicator of male reproductive status in Kangayam breed cattle.

Kangayam cattle are one of the drought breeds in India with distinct attributes. Agricultural transformation has led to a decline in many pure-breed indigenous cattle, including the Kangayam breed. Hence, a study on the reproductive physiology of male Kangayam breed cattle is necessary to disentangle problems in the area of livestock improvement. In this study, we investigated the relationship between serum hormones and bio-constituents and ascertained the potential of saliva as an indicator of the reproductive status of Kangayam cattle (Bos indicus). The present study confirms that cholesterol was higher in intact males and lower in prepubertal and castrated males. Testosterone levels were also higher in intact males than in castrated or prepubertal males. Hence, it can be inferred that high cholesterol levels contribute to active derivatization of testosterone in intact males. In contrast, reduced cholesterol availability leads to decreased testosterone synthesis in castrated and prepubertal males. Furthermore, it is reasonable to speculate that testosterone could have influenced salivary fern patterns in intact males, and thus, fern-like crystallization in the saliva was apparent. The unique salivary compounds identified through GC-MS across various reproductive statuses of Kangayam males may advertise their physiological status to conspecifics. In addition, the presence of odorant-binding protein (OBP) in saliva further supports its role in olfactory communication. This study attested to a posssible interlink between gonadal status and serum biochemical profiles. The salivary fern pattern revealed in this study can be used as a predictive tool, and the presence of putative volatiles and OBP adds evidence to the role of saliva in chemical communication.

Salivary metabolites are promising noninvasive biomarkers of drug-induced liver injury.

BACKGROUND: Drug-induced liver injury (DILI) is one of the most common adverse events of medication use, and its incidence is increasing. However, early detection of DILI is a crucial challenge due to a lack of biomarkers and noninvasive tests. AIM: To identify salivary metabolic biomarkers of DILI for the future development of noninvasive diagnostic tools. METHODS: Saliva samples from 31 DILI patients and 35 healthy controls (HCs) were subjected to untargeted metabolomics using ultrahigh-pressure liquid chromatography coupled with tandem mass spectrometry. Subsequent analyses, including partial least squares-discriminant analysis modeling, t tests and weighted metabolite coexpression network analysis (WMCNA), were conducted to identify key differentially expressed metabolites (DEMs) and metabolite sets. Furthermore, we utilized least absolute shrinkage and selection operato and random fores analyses for biomarker prediction. The use of each metabolite and metabolite set to detect DILI was evaluated with area under the receiver operating characteristic curves. RESULTS: We found 247 differentially expressed salivary metabolites between the DILI group and the HC group. Using WMCNA, we identified a set of 8 DEMs closely related to liver injury for further prediction testing. Interestingly, the distinct separation of DILI patients and HCs was achieved with five metabolites, namely, 12-hydroxydodecanoic acid, 3-hydroxydecanoic acid, tetradecanedioic acid, hypoxanthine, and inosine (area under the curve: 0.733-1). CONCLUSION: Salivary metabolomics revealed previously unreported metabolic alterations and diagnostic biomarkers in the saliva of DILI patients. Our study may provide a potentially feasible and noninvasive diagnostic method for DILI, but further validation is needed.

The use of bioluminescent enzyme bioassay for the analysis of human saliva: Advantages and disadvantages.

The purpose of the work was to find optimal conditions for bioluminescent enzymatic analysis of saliva (based on the use of NADH:FMN oxidoreductase + luciferase) and then to determine the biological effect of using bioluminescence assay of saliva to study the physiological state of the body under normal and pathological conditions. The saliva of snowboarders and students were studied in the "rest-training" model. The saliva of patients diagnosed with acute pharyngitis was examined in the "sick-healthy" model. Bioluminescence assay was performed with a lyophilized and immobilized bi-enzyme system using cuvette, plate, and portable luminometers. The concentrations of secretory immunoglobulin A (sIgA) and cortisol were determined by enzyme immunoassay, and the total protein content was measured by spectrophotometric method. The activity of the bioluminescent system enzymes increased as the amount and volume of saliva in the sample was decreased. The cuvette and plate luminometers were sensitive to changes in the luminescence intensity in saliva assay. Luminescence intensity correlated with the concentrations of sIgA and cortisol. The integrated bioluminescent index for saliva was reduced in the "rest-training" model and increased in the "sick-healthy" model. Thus, the non-invasive bioluminescent saliva analysis may be a promising tool for assessing the health of the population.

Assessment of severity and mineral composition of saliva in schoolchildren with molar-incisor hypomineralization (MIH).

The aim of the study was to evaluate the severity of molar incisor hypomineralisation (MIH), related oral health and investigate salivary mineral composition. The study was conducted with 50 participants aged between 6-15 years who were effected with MIH and 50 without MIH. The International Caries Detection and Assessment System (ICDAS) scores, Decayed, Missing, Filled Teeth/Surface (DMFT/S), dft/s and gingival/plaque indices were evaluated. The pH, flow rate, buffering capacity and mineral composition of saliva was measured. "Student t" test, one-way analysis of variance in repeated measurements of groups, and Tukey multiplex in subgroup comparisons was used. Kruskal-Wallis, Mann-Whitney U, Wilcoxon and chi-square tests were used to analyze qualitative data and compare groups. A total of 100 children (57 females 43 males, mean age 10.12 ± 1.85) participated in the study. There was no difference between ICDAS, DMFT/S scores, but dft/s index values were statistically significant (p = 0.001). The simplified oral hygiene index of MIH patients were statistically higher, but no significant differences were found in modified gingival indices (p = 0.52). Although the salivary pH and flow rate of the patients in the study group were lower, the buffering capacity was higher than those in the control group, but no significant difference was observed (p = 0.64). The mean values of phosphorus, carbon and calcium content in the saliva samples of MIH patients were higher than those of patients without MIH, and this difference was low for phosphorus (p = 0.76) and carbon (p = 0.74), but significantly higher for calcium. To the best of our knowledge, this is the first study to evaluate the association between calcium, phosphate and carbon levels in saliva of children with MIH. The significantly high amount of calcium in the saliva of patients with MIH suggests that further investigations are needed.

Raman hyperspectroscopy of saliva and machine learning for Sjögren's disease diagnostics.

Sjögren's disease is an autoimmune disorder affecting exocrine glands, causing dry eyes and mouth and other morbidities. Polypharmacy or a history of radiation to the head and neck can also lead to dry mouth. Sjogren's disease is often underdiagnosed due to its non-specific symptoms, limited awareness among healthcare professionals, and the complexity of diagnostic criteria, limiting the ability to provide therapy early. Current diagnostic methods suffer from limitations including the variation in individuals, the absence of a single diagnostic marker, and the low sensitivity and specificity, high cost, complexity, and invasiveness of current procedures. Here we utilized Raman hyperspectroscopy combined with machine learning to develop a novel screening test for Sjögren's disease. The method effectively distinguished Sjögren's disease patients from healthy controls and radiation patients. This technique shows potential for development of a single non-invasive, efficient, rapid, and inexpensive medical screening test for Sjögren's disease using a Raman hyper-spectral signature.

Prolongation of the gastric residence time of caffeine after administration in fed state: Comparison of effervescent granules with an extended release tablet.

The aim of the present study was to investigate the gastroretentive capacity of different formulation principles. This was indirectly determined by the absorption behavior of caffeine from the dosage forms. A slow and continuous appearance of caffeine in the saliva of healthy volunteers was used as a parameter for a prolonged gastric retention time. For this purpose, a four-way study was conducted with twelve healthy volunteers using the following test procedures: (1) Effervescent granules with 240 mL of still water administered in fed state, (2) effervescent granules with 20 mL of still water in fed state, (3) extended release (ER) tablet with 240 mL of still water in fed state, and (4) effervescent granules with 240 mL of still water in fasted state. The initial rise of the caffeine concentrations was more pronounced after the intake of the effervescent granules in the fed state compared to that of the ER tablets. However, tmax tended to be shorter in the fed study arms following administration of the ER tablet compared to the granules. Overall, the application of active pharmaceutical ingredients formulated as effervescent granules seems to be a promising approach to increase their gastric residence time after intake in fed state.

Effects of noise exposure on stress hormone changes during task performance in young Korean men: quasi-experimental study.

Numerous studies have suggested that noise exposure might be associated with changes in stress hormone levels. However, quantitative evidence for these effects in humans is rare and remains controversial. This study aimed to investigate the acute effects of exposure to noise and its different levels on stress hormone changes in task performance. Quasi-experimental noise exposure environment was established for 90 male university student volunteers in their twenties, and each was exposed to different noise levels during task performance. The stress hormones tested included cortisol, adrenocorticotropic hormone (ACTH), adrenaline, and noradrenaline. A one-way ANOVA was performed to investigate differences in hormone levels measured in the three groups according to the noise exposure levels (35, 45, or 75 dB). Analysis of covariance (ANCOVA) was used to adjust for confounding factors that might affect hormone levels. After adjusting for confounders, significant exposure-dependent differences were found in hormone levels in salivary cortisol, serum cortisol, serum ACTH, and serum adrenaline. The amount of hormonal increase in 75 dB exposure group compared to 35 or 45 dB groups was detected. Similar results were also seen in the rate of change analysis. Our findings indicate that short-term noise exposure during task performance elevates stress hormone levels. Further, the extent of stress hormone alterations varies with noise exposure levels. Changes in hormone levels are an objective measure that may be used to identify health effects and stress responses in various noise environments.

Salivary analysis to unveil the paradigma of stress of domestic horses reared in the wild.

Horse welfare is the product of multiple factors, including behavioral and physiological adjustments to cope with stressful situation regarding environment and housing condition. Collectively, it is supposed that a horse kept in the wild has a lower level of stress than other housing system, and the aim of the present study was to investigate the level of stress in domestic horses reared in the wild and then moved to human controlled housing, through saliva analysis. Twelve clinically healthy Catria (Italian local breed) mares, usually reared in the wild, were moved into collective paddocks for a folkloric event. Saliva samples were obtained before and after the change of housing condition to evaluate stress biomarkers including salivary cortisol, salivary alpha-amylase, and butyrylcholinesterase (BChol). The mares were also scored using the Welfare Aggregation and Guidance (WAG) Tool to highlight the presence of abnormal behaviors. Despite the absence of differences in behavioral scores between wild and paddocks, salivary cortisol and BChol were found to be higher in the wild and lower when mares were moved to paddocks. The highest concentrations in stress biomarkers like salivary cortisol and BChol in the wild was unexpected, but the need for managing hierarchical relationships, and the exposure to feral animals, predators, and weather changes, might explain these findings. The overall results of the present study may provide further knowledge toward stress response in domesticated horses living in the wild moved to human controlled housing system.

Salivary, acidic and enzymatic degradation of resin composite subjected to different finishing and polishing systems.

PURPOSE: To evaluate the effect of different finishing and polishing systems on the surface roughness of a resin composite subjected to simulated saliva-, acid-, and enzyme-induced degradation. METHODS: 160 specimens (n= 40) were fabricated with Filtek Z350 XT nanofilled composite and analyzed for average surface roughness (Ra). The specimens were finished and polished using: AD - Al2O3-impreginated rubberized discs (medium, fine, and superfine grit, Sof-Lex); SD - silicon carbide and Al2O3-impregnated rubberized discs (coarse, medium and fine grit, Jiffy,); MB - 12- and 30-multiblade burs. The control group (CT) (n= 40) comprised specimens with a Mylar-strip-created surface. Specimens from each group were immersed in 1 mL of one of the degradation methods (n= 10): artificial saliva (ArS: pH 6.75), cariogenic challenge (CaC: pH 4.3), erosive challenge (ErC: 0.05M citric acid, pH 2.3) or enzymatic challenge (EzC: artificial saliva with 700 µg/mL of albumin, pH 6.75). The immersion period simulated a time frame of 180 days. Ra measurements were also performed at the post-polishing and post-degradation time points. The data were evaluated by three-way ANOVA for repeated measures and the Tukey tests. RESULTS: There was significant interaction between the finishing/polishing system and the degradation method (P= 0.001). AD presented the greatest smoothness, followed by SD. After degradation, CT, AD and SD groups became significantly rougher, but not the MB group, which presented no difference in roughness before or after degradation. CT and AD groups showed greater roughness in CaC, ErC and EzC than in ArS. The SD group showed no difference in roughness when the specimens were polished with CaC, EzC or ArS, but those treated with ErC had greater roughness. In the MB group, the lower roughness values were found after using CaC and EzC, while the higher values were found using ErC or ArS. CLINICAL SIGNIFICANCE: As far as degradation resistance of nanofilled composite to hydrolysis, bacterial and dietary acids and enzymatic reactions is concerned, restorations that had been finished and polished with Al2O3-impregnated discs had the smoothest surfaces.

Unique and interactive effects of threat and deprivation on latent trait cortisol among emerging adults.

Though considerable work supports the Dimensional Model of Adversity and Psychopathology, prior research has not tested whether the dimensions-threat (e.g., abuse) and deprivation (e.g., neglect)-are uniquely related to salivary trait indicators of hypothalamic pituitary adrenal (HPA) axis activity. We examined the unique and interactive effects of threat and deprivation on latent trait cortisol (LTC)-and whether these effects were modified by co-occurring adversities. Emerging adults (n = 90; Mage = 19.36 years; 99.88% cisgender women) provided salivary cortisol samples four times a day (waking, 30 min and 45 min postwaking, bedtime) over three 3-day measurement waves over 13 weeks. Contextual life stress interviews assessed early adversity. Though the effects varied according to the conceptualization of early adversity, overall, threat-but not deprivation, nor other co-occurring adversities-was uniquely associated with the across-wave LTC. Specifically, the incidence and frequency of threat were each negatively related to the across-wave LTC. Threat severity was also associated with the across-wave LTC, but only among those with no deprivation. Finally, the effects of threat were modified by other co-occurring adversities. Findings suggest that threat has unique implications for individual differences in HPA axis activity among emerging adults, and that co-occurring adversities modify such effects.

Female RNA concussion (FeRNAC) study: assessing hormone profiles and salivary RNA in females with concussion by emergency departments in New Zealand: a study protocol.

BACKGROUND: Females of reproductive age with concussion report a greater number of symptoms that can be more severe and continue for longer than age matched males. Underlying mechanisms for sex differences are not well understood. Short non-coding Ribonucleic Acids (sncRNAs) are candidate salivary biomarkers for concussion and have been studied primarily in male athletes. Female sex hormones influence expression of these biomarkers, and it remains unclear whether a similar pattern of sncRNA expression would be observed in females following concussion. This study aims to evaluate recovery time, the ratio of salivary sncRNAs and symptom severity across different hormone profiles in females presenting to emergency departments (ED) with concussion and, to investigate the presence of low energy availability (LEA) as a potential modifier of concussion symptoms. METHODS: This prospective cohort study recruits participants from New Zealand EDs who are biologically female, of reproductive age (16-50 years) and with a confirmed diagnosis of concussion from an ED healthcare professional. Participants are excluded by ED healthcare professionals from study recruitment as part of initial routine assessment if they have a pre-diagnosed psychiatric condition, neurological condition (i.e., epilepsy, cerebral palsy) or more than three previously diagnosed concussions. Participants provide a saliva sample for measurement of sncRNA's, and online survey responses relating to hormone profile and symptom recovery at 7-day intervals after injury until they report a full return to work/study. The study is being performed in accordance with ethical standards of the Declaration of Helsinki with ethics approval obtained from the Health and Disability Ethics Committee (HDEC #2021 EXP 11655), Auckland University of Technology Ethics Committee (AUTEC #22/110) and locality consent through Wellington hospital research office. DISCUSSION: If saliva samples confirm presence of sncRNAs in females with concussion, it will provide evidence of the potential of saliva sampling as an objective tool to aid in diagnosis of, and confirmation of recovery from, concussion. Findings will determine whether expression of sncRNAs is influenced by steroid hormones in females and may outline the need for sex specific application and interpretation of sncRNAs as a clinical and/or research tool. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry (ANZCTR) registration number ACTRN12623001129673.

Laser-printed paper ELISA and hydroxyapatite immobilization for colorimetric congenital anomalies screening in saliva.

BACKGROUND: Alpha-fetoprotein (AFP) is a fetal protein that can indicate congenital anomalies such as Down syndrome and spinal canal blockage when detected at abnormal levels in pregnant women. Current AFP detection methods rely on invasive blood or serum samples, which require sophisticated equipment. From the many solutions proposed, colorimetric paper-based assays excel in point-of-care settings. The concept of paper-based ELISA (p-ELISA) enhances traditional methods, aligning with the ASSURED criteria for diagnostics in resource-limited regions. Despite success in microfluidic paper-based assay devices, laser printing remains underexplored for p-ELISA. Additionally, modifying the paper surface provides an additional layer of sensitivity enhancement. RESULTS: In this study, we developed a novel laser-printed paper-based ELISA (LP-pELISA) for rapid, sensitive, and noninvasive detection of AFP in saliva samples. The LP-pELISA platform was fabricated by printing hydrophobic barriers on filter paper using a laser printer, followed by depositing hydroxyapatite (HAp) as an immobilization material for the antibodies. The colorimetric detection was achieved using AuNPs functionalized with anti-AFP antibodies and silver nitrate enhancement. The LP-pELISA exhibited a linear response for AFP detection in both buffer and saliva samples over a range of 1.0-800 ng mL-1, with a limit of detection (LOD) reaching 1.0 ng mL-1. The assay also demonstrated good selectivity, repeatability, reproducibility, and stability. The LP-pELISA was further validated by testing spiked human saliva samples, showing its potential for point-of-care diagnosis of congenital disabilities. SIGNIFICANCE: The LP-pELISA is a noninvasive platform showcasing simplicity, cost-effectiveness, and user-friendliness, utilizing laser printing, hydroxyapatite modification, and saliva samples to efficiently detect AFP. Beyond its application for AFP, this method's versatility extends to other biomarkers, positioning it as a catalyst for the evolution of paper-based biosensors. The LP-pELISA holds promise as a transformative tool for point-of-care diagnostics, fostering advancements in healthcare with its innovative technology.

Evaluation of salivary stress markers and inflammatory cytokine levels in peri-implantitis patients.

BACKGROUND AND OBJECTIVE: Psychological stress has been identified in some observational studies as a potential factor that may modify and affect periodontal diseases, but there are no similar data for peri-implantitis. The aim of this study was to determine the relationship between interleukin (IL)-1ß, IL-6, IL-10, interferon (IFN)α inflammatory cytokines and the psychological stress-related markers, glucocorticoid receptor-α (GRα), and salivary α-amylase (sAA) gene expression levels in saliva samples obtained from healthy implants and peri-implantitis patients. MATERIALS AND METHODS: The study included a total of 50 systemically healthy subjects. Peri-implant clinical parameters were recorded and psychological stress level was evaluated with the hospital anxiety and depression scale (HAD) and state-trait anxiety inventory (STAI) questionnaire forms. Following the evaluations, the patients were divided into 4 groups according their stress and clinical status (Ia, Ib, IIa, IIb). IL-1ß, IL-6, IL-10, IFNα, GRα, sAA gene expression levels in the saliva samples were quantified by quantitative polymerase chain reaction (qPCR). RESULTS: In the group of peri-implantitis who had a high score in stress level assessment scales, significantly higher IL-1ß, IL-6, sAA expression levels were observed (p < 0.001). The IL-10 gene expression levels were lower in the groups with a high score in the stress level assessment scales (p < 0.001). GRα gene was expressed at lower levels in the group of peri-implantitis who had a high score in stress level assessment scales but the difference was not statistically significant (p = 0.065). CONCLUSION: The study findings suggest that psychological stress may increase the inflammation associated with peri-implantitis by affecting cytokine expression levels. CLINICAL RELEVANCE: To prevent peri-implantitis or reduce its prevalence, it could be beneficial to evaluate stress levels and identify individuals experiencing stress.

Salivary Proteinase 3 as a Biomarker for Caries Severity in Children: A Cross-sectional Study.

AIM: This study aims to evaluate the relation between salivary proteinase 3 (PR3) concentration and caries severity in children. MATERIALS AND METHODS: Six-to-eight-year age group children, from the Outpatient Department of Pediatric and Preventive dentistry at PMS Dental College were selected for the study. From these children, three groups each consisting of 28 children were selected according to the dental caries severity. Three groups were: (1) No Dental Caries group, (2) Low Dental Caries group with DMFT/DEFT score of 1-4, and (3) High Dental Caries group with DMFT/DEFT score of 5-15. Thus, a total of 84 children who satisfied the inclusion criteria were selected. The concentration of PR3 in saliva of the donors were analyzed using an ELISA kit. One way ANOVA was used for finding the relation of salivary PR3 concentration with caries severity. Pairwise comparison of PR3 concentration and caries severity were analyzed using post hoc Tukey test. RESULTS: Severity of caries and concentration of salivary PR3 showed an inverse relation. As the caries severity increases there was a decrease in PR3 concentration and vice versa. CONCLUSION: The children with high caries severity showed lower concentration of PR3 in their saliva compared with those with lower caries severity which indicates that PR3 can be used as a biomarker for assessing caries severity and also paves way to use PR3 as a caries vaccine in future. Nowadays, interest toward noninvasive and personalized dentistry has been increased. Molecular assays using salivary biomarkers can be an effective tool in detecting the caries in earlier stages and assessing a patient's caries risk. CLINICAL SIGNIFICANCE: Salivary PR3 can be used as prognostic biomarker for assessing caries severity and after treatment the value of PR3 can be used as a assessment tool to confirm its relation with caries. How to cite this article: Karthika S, George S, Soman A, et al. Salivary Proteinase 3 as a Biomarker for Caries Severity in Children: A Cross-sectional Study. J Contemp Dent Pract 2024;25(3):236-240.

Turn-off enzyme activity of histidine-rich peptides for the detection of lysozyme.

An assay that integrates histidine-rich peptides (HisRPs) with high-affinity aptamers was developed enabling the specific and sensitive determination of the target lysozyme. The enzyme-like activity of HisRP is inhibited by its interaction with a target recognized by an aptamer. In the presence of the target, lysozyme molecules progressively assemble on the surface of HisRP in a concentration-dependent manner, resulting in the gradual suppression of enzyme-like activity. This inhibition of HisRP's enzyme-like activity can be visually observed through color changes in the reaction product or quantified using UV-visible absorption spectroscopy. Under optimal conditions, the proposed colorimetric assay for lysozyme had a detection limit as low as 1 nM and exhibited excellent selectivity against other nonspecific interferents. Furthermore, subsequent research validated the practical applicability of the developed colorimetric approach to saliva samples, indicating that the assay holds significant potential for the detection of lysozymes in samples derived from humans.