Oral Antiviral Defense: Saliva- and Beverage-like Hypotonicity Dynamically Regulate Formation of Membraneless Biomolecular Condensates of Antiviral Human MxA in Oral Epithelial Cells.

The oral mucosa represents a defensive barrier between the external environment and the rest of the body. Oral mucosal cells are constantly bathed in hypotonic saliva (normally one-third tonicity compared to plasma) and are repeatedly exposed to environmental stresses of tonicity, temperature, and pH by the drinks we imbibe (e.g., hypotonic: water, tea, and coffee; hypertonic: assorted fruit juices, and red wines). In the mouth, the broad-spectrum antiviral mediator MxA (a dynamin-family large GTPase) is constitutively expressed in healthy periodontal tissues and induced by Type III interferons (e.g., IFN-λ1/IL-29). Endogenously induced human MxA and exogenously expressed human GFP-MxA formed membraneless biomolecular condensates in the cytoplasm of oral carcinoma cells (OECM1 cell line). These condensates likely represent storage granules in equilibrium with antivirally active dispersed MxA. Remarkably, cytoplasmic MxA condensates were exquisitely sensitive sensors of hypotonicity-the condensates in oral epithelium disassembled within 1-2 min of exposure of cells to saliva-like one-third hypotonicity, and spontaneously reassembled in the next 4-7 min. Water, tea, and coffee enhanced this disassembly. Fluorescence changes in OECM1 cells preloaded with calcein-AM (a reporter of cytosolic "macromolecular crowding") confirmed that this process involved macromolecular uncrowding and subsequent recrowding secondary to changes in cell volume. However, hypertonicity had little effect on MxA condensates. The spontaneous reassembly of GFP-MxA condensates in oral epithelial cells, even under continuous saliva-like hypotonicity, was slowed by the protein-phosphatase-inhibitor cyclosporin A (CsA) and by the K-channel-blocker tetraethylammonium chloride (TEA); this is suggestive of the involvement of the volume-sensitive WNK kinase-protein phosphatase (PTP)-K-Cl cotransporter (KCC) pathway in the regulated volume decrease (RVD) during condensate reassembly in oral cells. The present study identifies a novel subcellular consequence of hypotonic stress in oral epithelial cells, in terms of the rapid and dynamic changes in the structure of one class of phase-separated biomolecular condensates in the cytoplasm-the antiviral MxA condensates. More generally, the data raise the possibility that hypotonicity-driven stresses likely affect other intracellular functions involving liquid-liquid phase separation (LLPS) in cells of the oral mucosa.

In vitro simulated saliva, gastric, and intestinal digestion followed by faecal fermentation reveals a potential modulatory activity of Epimedium on human gut microbiota.

Herba Epimedii, known for its rich array of bioactive ingredients and widespread use in ethnopharmacological practices, still lacks a comprehensive understanding of its gastrointestinal biotransformation. In this study, we qualitatively explored the dynamic changes in Epimedium sagittatum components during in vitro simulated digestions, with a quantitative focus on its five major flavonoids. Notably, significant metabolism of E. sagittatum constituents occurred in the simulated small intestinal fluid and colonic fermentation stages, yielding various low molecular weight metabolites. Flavonoids like kaempferol glycosides were fully metabolized in the simulated intestinal fluid, while hyperoside digestion occurred during simulated colon digestion. Colonic fermentation led to the production of two known bioactive isoflavones, genistein, and daidzein. The content and bioaccessibility of the five major epimedium flavonoids-icariin, epimedin A, epimedin B, epimedin C, and baohuoside I-significantly increased after intestinal digestion. During colon fermentation, these components gradually decreased but remained incompletely metabolized after 72 h. Faecal samples after E. sagittatum fermentation exhibited shift towards dominance by Lactobacillus (Firmicutes), Bifidobacterium (Actinobacteria), Streptococcus (Firmicutes), and Dialister (Firmicutes). These findings enhance our comprehension of diverse stages of Herba Epimedii constituents in the gut, suggesting that the primary constituents become bioaccessible in the colon, where new bioactive compounds may emerge.

Machine learning-based detection of acute psychosocial stress from body posture and movements.

Investigating acute stress responses is crucial to understanding the underlying mechanisms of stress. Current stress assessment methods include self-reports that can be biased and biomarkers that are often based on complex laboratory procedures. A promising additional modality for stress assessment might be the observation of body movements, which are affected by negative emotions and threatening situations. In this paper, we investigated the relationship between acute psychosocial stress induction and body posture and movements. We collected motion data from N = 59 individuals over two studies (Pilot Study: N = 20, Main Study: N = 39) using inertial measurement unit (IMU)-based motion capture suits. In both studies, individuals underwent the Trier Social Stress Test (TSST) and a stress-free control condition (friendly-TSST; f-TSST) in randomized order. Our results show that acute stress induction leads to a reproducible freezing behavior, characterized by less overall motion as well as more and longer periods of no movement. Based on these data, we trained machine learning pipelines to detect acute stress solely from movement information, achieving an accuracy of 75.0 ± 17.7 % (Pilot Study) and 73.4 ± 7.7 % (Main Study). This, for the first time, suggests that body posture and movements can be used to detect whether individuals are exposed to acute psychosocial stress. While more studies are needed to further validate our approach, we are convinced that motion information can be a valuable extension to the existing biomarkers and can help to obtain a more holistic picture of the human stress response. Our work is the first to systematically explore the use of full-body body posture and movement to gain novel insights into the human stress response and its effects on the body and mind.

Expression analysis of zinc-metabolizing enzymes in the saliva as a new method of evaluating zinc content in the body: two case reports and a review of the literature.

BACKGROUND: The activity level of alkaline phosphatase, a zinc-requiring enzyme in the serum, is used to indicate zinc nutritional status; however, it does not correlate with serum zinc levels or subjective symptoms of taste disorder in many cases. Hence, this study focused on the total activity of alkaline phosphatase, a zinc-requiring enzyme. The total alkaline phosphatasa activity level in the saliva was measured before and after zinc supplementation, and the results were compared with serum zinc levels. CASE PRESENTATION: This study included patients with hypozincemia, specifically a patient with zinc-deficient taste disorder (patient 1: a 69-year-old Japanese woman) and a patient with glossodynia with zinc deficiency (patient 2: an 82-year-old Japanese woman). Saliva samples were collected, and blood tests were performed before and after zinc supplementation. Subjective symptoms and serum zinc levels were simultaneously evaluated. Zinc supplementation was performed using zinc acetate hydrate or Polaprezinc. CONCLUSIONS: Total alkaline phosphatase activity levels were found to be associated with serum zinc levels and subjective symptoms. A further study with a higher number of patients is necessary to confirm whether total alkaline phosphatase activity levels more accurately reflect the amounts of zinc in the body than serum zinc levels.

Effect of prepared herbal mouthwash in maintaining the oral health of school children: A single-blind randomised control trial.

BACKGROUND AND AIM: Oral health is a vital indicator of overall well-being, quality of life, and general health, with historical roots in Unani medicine (i.e. preventive measures, oral hygiene, and treatment). This study aims to assess the efficacy and safety of prepared herbal mouthwash in maintaining oral health among school children. EXPERIMENTAL PROCEDURE: 110 schoolchildren were randomly assigned to the test group (treated with herbal mouthwash -Anacyclus pyrenthrum DC, Punica granutum (pericarp), Capparis spinosa (root bark), and Quercus infectoria Oliv (galls)) and the control group (treated with 0.2 % Chlorhexidine Mouthwash) for 30-days. The response was assessed by DMFT, Salivary pH, Oral hygiene index-simplified (OHI-S), Plaque index (Loe & Silness) (PI), Plaque index simplified (O'Leary et al., 1972) (PI-S), Gingival Index (Löe-Silness) (GI), and Bleeding on probing (BOP) on baseline and 30th day GI and PI were further assessed on 60th, 90th and 120th days to determine the sustainable effect of the intervention. RESULTS: After treatment, Mean OHI-S, PI, PI-S, GI and BOP significantly reduced on the 30th day from baseline (p ≤ 0.001) in both groups. After discontinuing intervention, both groups showed significantly lower PI and GI scores on the 120th day from baseline. (p < 0.001). CONCLUSION: This study revealed that prepared mouthwash is safe and significantly effective in maintaining oral health, and it could be used as an adjunct to mechanical oral hygiene measures.

Evaluation of photobiomodulation in the salivary production of patients with hyposalivation induced by antihypertensive drugs - A blind, randomized, controlled clinical trial.

BACKGROUND: Arterial hypertension is a systemic condition that affects about 35% of the world population. The drugs that are used for its control can produce hyposalivation. This work evaluated the effect of photobiomodulation on salivary flow rate, salivary pH, total protein concentration, and calcium concentration in individuals using antihypertensive medications. MATERIAL AND METHODS: 41 subjects were randomly allocated in one of two groups: control (placebo) and photobiomodulation. The subjects had their salivary glands (20 sites) irradiated with a laser emitting at 808 nm, 4J/site once a week for 4 weeks and had their salivary flow measured before and after the whole treatment. RESULTS: The intragroup analysis (before and after treatment) shows a significant difference for both non-stimulated and stimulated salivary flow in the photobiomodulation group (p = 0.0007 and p = 0.0001, respectively). Comparing the placebo with the photobiomodulation group, significant differences were found for both non-stimulated (p = 0.0441) and stimulated salivary flow (p = 0.0441) after the treatment. No significant differences were found in pH, total protein concentration, calcium concentration. CONCLUSION: Despite the usage of drugs that influence the nervous system and typically result in a reduction of saliva production, photobiomodulation demonstrated a remarkable ability to enhance saliva production by a significant 75%.

Increased nitrate intake from beetroot juice over 4 weeks affects nitrate metabolism, but not vascular function or blood pressure in older adults with hypertension.

The decline in vascular function and increase in blood pressure with aging contribute to an increased cardiovascular disease risk. In this randomized placebo-controlled crossover study, we evaluated whether previously reported cardiovascular benefits of plant-derived inorganic nitrate via nitric oxide (NO) translate into improved vascular function and blood pressure-lowering in 15 men and women (age range: 56-71 years) with treated hypertension. We investigated the effects of a single ∼400 mg-dose at 3 hours post-ingestion (3H POST) and the daily consumption of 2 × âˆ¼400 mg of nitrate through nitrate-rich compared with nitrate-depleted (placebo) beetroot juice over 4 weeks (4WK POST). Measurements included nitrate and nitrite in plasma and saliva; endothelial-dependent and -independent forearm blood flow (FBF) responses to acetylcholine (FBFACh) and glyceryltrinitrate (FBFGTN); and clinic-, home- and 24-hour ambulatory blood pressure. Compared to placebo, plasma and salivary nitrate and nitrite increased at 3H and 4WK POST following nitrate treatment (P < 0.01), suggesting a functioning nitrate-nitrite-NO pathway in the participants of this study. There were no differences between treatments in FBFACh and FBFGTN-area under the curve (AUC) ratios [AUC ratios after (3H POST, 4WK POST) compared with before (PRE) the intervention], or 24-hour ambulatory blood pressure or home blood pressure measures (P > 0.05). These findings do not support the hypothesis that an increased intake of dietary nitrate exerts sustained beneficial effects on FBF or blood pressure in hypertensive older adults, providing important information on the efficacy of nitrate-based interventions for healthy vascular aging. This study was registered under ClinicialTrials.gov (NCT04584372).

Black Phosphorus-Based Reusable Biosensor Platforms for the Ultrasensitive Detection of Cortisol in Saliva.

A black phosphorus (BP)-based reusable biosensor platform is developed for the repeated and real-time detection of cortisol using antibody-conjugated magnetic particle (MP) structures as a refreshable receptor. Here, we took advantage of the low-noise characteristics of a mechanically exfoliated BP-based field-effect transistor (FET) and hybridized it with anti-cortisol antibody-functionalized MPs to build a highly sensitive cortisol sensor. This strategy allowed us to detect cortisol down to 1 aM in real time and discriminate cortisol from other hormones. In this case, we could easily remove MPs with used antibodies from the surface of a BP-FET and reuse the chip for up to eight repeated sensing operations. Moreover, since our platform could be fabricated using conventional photolithography techniques and the sensor can be reused multiple times, one should be able to significantly reduce operation costs for practical applications. Furthermore, this method could be utilized to detect different hormones with high sensitivity and selectivity in complex environments such as artificial saliva solutions. In this respect, our reusable BP-FET biosensing platform can be a powerful tool for versatile applications such as clinical diagnosis and basic biological analysis by conjugating various antibodies.

Evaluation of the Canadian approved drug screening equipment cut-off levels for tetrahydrocannabinol (THC).

In 2018, Canada introduced roadside oral fluid (OF) screening devices, called Approved Drug Screening Equipment (ADSE), as an investigative tool in impaired driving investigations to detect tetrahydrocannabinol (THC), cocaine and/or methamphetamine in drivers. In this work, we compare the detection and concentration of THC in blood samples collected from suspected impaired drivers that tested positive at the roadside for THC on an ADSE. The two ADSEs that were utilized were the Dräger DrugTest® 5000 (DDT) and the Abbott SoToxa™ (SoToxa), both configured with a THC OF concentration cut-off concentration of 25 ng/mL. Blood samples were screened for cannabinoids using immunoassay and positive results were followed up by confirmation/quantitation of THC by ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS-MS). A total of 230 cases were available where a blood sample was collected from a suspected impaired driver subsequent to a positive THC screen result on an ADSE. The blood samples were taken an average of 1.4 hours (range = 9 minutes to 3.2 hours) after the ADSE test. THC was confirmed in 98% of blood samples with concentrations across all samples ranging from not detected (cut = off 0.5 ng/mL) to greater than 20 ng/mL. Further, 90% of the blood samples had a THC concentration of 2.0 ng/mL (the lower per se limit in Canada) or greater. A positive ADSE test of a suspected impaired driver may predict that the driver has a detectable level of THC in their blood, and there is a high likelihood that the THC blood concentration is 2.0 ng/mL or higher. Hence, ADSE may be a useful tool for law enforcement and aid in the development of grounds to believe that a driver is operating a conveyance with a THC concentration exceeding Canadian per se limits.

Human saliva modifies growth, biofilm architecture, and competitive behaviors of oral streptococci.

The bacteria within supragingival biofilms participate in complex exchanges with other microbes inhabiting the same niche. One example is the mutans group streptococci (Streptococcus mutans), implicated in the development of tooth decay, and other health-associated commensal streptococci species. Previously, our group transcriptomically characterized intermicrobial interactions between S. mutans and several species of oral bacteria. However, these experiments were carried out in a medium without human saliva. To better mimic their natural environment, we first evaluated how inclusion of saliva affected growth and biofilm formation of eight Streptococcus species individually and found saliva to positively benefit growth rates while negatively influencing biofilm biomass accumulation and altering spatial arrangement. These results carried over during evaluation of 29 saliva-derived isolates of various species. Surprisingly, we also found that addition of saliva increased the competitive behaviors of S. mutans in coculture competitions against commensal streptococci that led to increases in biofilm microcolony volumes. Through transcriptomically characterizing mono- and cocultures of S. mutans and Streptococcus oralis with and without saliva, we determined that each species developed a nutritional niche under mixed-species growth, with S. mutans upregulating carbohydrate uptake and utilization pathways while S. oralis upregulated genome features related to peptide uptake and glycan foraging. S. mutans also upregulated genes involved in oxidative stress tolerance, particularly manganese uptake, which we could artificially manipulate by supplementing in manganese leading to an advantage over its opponent. Our report highlights observable changes in microbial behaviors through leveraging environmental- and host-supplied resources over their competitors. IMPORTANCE: Dental caries (tooth decay) is the most prevalent disease for both children and adults nationwide. Caries are initiated from demineralization of the enamel due to organic acid production through the metabolic activity of oral bacteria growing in biofilm communities attached to the tooth's surface. Mutans group streptococci are closely associated with caries development and initiation of the cariogenic cycle, which decreases the amount of acid-sensitive, health-associated commensal bacteria while selecting for aciduric and acidogenic species that then further drives the disease process. Defining the exchanges that occur between mutans group streptococci and oral commensals in a condition that closely mimics their natural environment is of critical need toward identifying factors that can influence odontopathogen establishment, persistence, and outgrowth. The goal of our research is to develop strategies, potentially through manipulation of microbial interactions characterized here, that prevent the emergence of mutans group streptococci while keeping the protective flora intact.

Concurrent Ingestion of Alkaline Water and L-Glutamine Enhanced Salivary α-Amylase Activity and Testosterone Concentration in Boxing Athletes.

Athletes often take sport supplements to reduce fatigue and immune disturbances during or after training. This study evaluated the acute effects of concurrent ingestion of alkaline water and L-glutamine on the salivary immunity and hormone responses of boxers after training. Twelve male boxing athletes were recruited in this study. During regular training, the participants were randomly divided into three groups and asked to consume 400 mL of alkaline water (Group A), 0.15 g/kg body weight of L-glutamine with 400 mL of water (Group G), and 0.15 g/kg of L-glutamine with 400 mL of alkaline water (Group A+G) at the same time each day for three consecutive weeks. Before and immediately after the training, saliva, heart rates, and the rate of perceived exertion were investigated. The activity of α-amylase and concentrations of lactoferrin, immunoglobulin A (IgA), testosterone, and cortisol in saliva were measured. The results showed that the ratio of α-amylase activity/total protein (TP) significantly increased after training in Group A+G but not in Group A or G, whereas the ratios of lactoferrin/TP and IgA/TP were unaffected in all three groups. The concentrations of salivary testosterone after training increased significantly in Group A+G but not in Group A or G, whereas the salivary cortisol concentrations were unaltered in all groups. In conclusion, concurrent ingestion of 400 mL of alkaline water and 0.15 g/kg of L-glutamine before training enhanced the salivary α-amylase activity and testosterone concentration of boxers, which would be beneficial for post-exercise recovery.

Salivary elemental signature of dental caries: a systematic review and meta-analysis of ionomics studies.

Trace- and macro-chemical elements are crucial for cellular physiological functioning, and their alterations in biological fluids might be associated with an underlying pathological state. Hence, this study aimed to examine and summarize the published literature concerning the application of salivary ionomics for caries diagnosis. An extensive search of studies was conducted using PubMed, EMBASE, Web of Science, and Scopus, without any language and year restriction for answering the following PECO question: "In subjects (i.e., children, adolescents, or adults) with good systematic health, are there any variations in the salivary concentrations of trace- or macro-elements between caries-free (CF) individuals and caries-active (CA) subjects?" A modified version of the QUADOMICS tool was used to assess the quality of the included studies. The Review Manager Version 5.4.1. was used for data analyses. The analysis of salivary chemical elements that significantly differed between CF and CA subjects was also performed. Thirty-four studies were included, involving 2299 CA and 1669 CF subjects, having an age range from 3 to 64 years in over 16 countries. The meta-analysis revealed a statistically significant difference (p < 0.05) in the salivary levels of calcium, phosphorus, chloride, magnesium, potassium, sodium, and zinc between CA and CF subjects, suggesting higher levels of calcium, phosphorus, potassium, and sodium in CF subjects while higher levels of chloride, magnesium, and zinc in CA patients. Half of the included studies (17/34) were considered high quality, while the remaining half were considered medium quality. Only zinc and chloride ions were found to be higher significantly and consistent in CF and CA subjects, respectively. Conflicting outcomes were observed for all other salivary chemical elements including aluminum, bromine, calcium, copper, fluoride, iron, potassium, magnesium, manganese, sodium, ammonia, nitrite, nitrate, phosphorus, lead, selenium, and sulfate ions.

Development and validation of a LC-MS/MS method for the quantification of phenolic compounds in human saliva after intake of a procyanidin-rich pine bark extract.

Plant-derived phenolic compounds are regularly ingested as food compounds or as food supplements. Concentrations of individual compounds and metabolites are typically measured in serum or urine samples. This, however, allows no conclusion on the distribution into organs and tissues. An easily accessible biofluid is saliva. At this point, it was not clear yet, whether polyphenols circulating in the blood would be secreted or diffuse into saliva. The purpose of the present study was to develop and validate a method using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) for analysis of phenolic compounds in human saliva. Method validation for the quantification of taxifolin, ferulic acid, caffeic acid, gallic acid, para-coumaric acid, and protocatechuic acid and the gut microbial catechin metabolite δ-(3,4-dihydroxyphenyl)-γ-valerolactone (M1) in human saliva was performed according to current guidelines for bioanalytical method validation. The lower limit of quantification ranged from 0.82 ng/ml for M1 to 8.20 ng/ml for protocatechuic acid. The method was successfully applied to an authentic saliva sample of a volunteer after swallowing of procyanidin-rich pine bark extract capsules (dietary supplement Pycnogenol®). All polyphenols except ferulic acid were quantified at concentrations ranging from 1.20 ng/ml (M1) to 10.34 ng/ml (gallic acid). Notably, in contrast to serum samples, all phenolic compounds were present without sulfate or glucuronic acid conjugation in saliva, suggesting an enzymatic deconjugation, e.g., by a ß-glucuronidase activity, during compound transfer from serum to saliva. Since M1 is only produced in the gut, its presence in saliva ruled out the possibility of sample contamination by phenolic compounds residing in the oral cavity after food intake. To the best of our knowledge, this is the first time that the gut microbiota-derived metabolite M1 has been detected in saliva. To further investigate the role of phenolic compounds in saliva, the described analytical method can be applied in clinical studies investigating the biodistribution of polyphenols and their metabolites.

The Photobiomodulation effect of 980 nm Diode Laser on Patients with Xerostomia.

PURPOSE: Xerostomia is not a disease in itself. This condition is associated with systemic diseases, medication use, and radiotherapy. Oral and general health suffer when the salivary gland´s function is reduced. The effectiveness of therapeutic methods in resolving this issue thus far has been inadequate. This study aimed to determine the photobiomodulation (PBM) effect of a 980 nm laser on xerostomia. METHODS: A quasi-experimental study was carried out using a 980 nm diode laser on 28 patients with xerostomia. A diode laser (980 nm) was used extortionally over the submandibular glands and parotids and it was used internally in the sublingual glands. The laser beam (extrorally 6 J, 300mW, 20 s, and introrally 4 J, 200mW, 20 s) was applied bilaterally every other day for 10 sessions. It should be noted that 8 points of the parotid, 2 points of the submandibular glands, and 2 points of the sublingual glands were irradiated. The saliva was collected each session using the Navazesh technique, 5 min before and after PBM. The Wilcoxon test was used to study the effect of laser therapy on saliva flow rate (volume and weight). The P < 0.05 level was considered a significant level. RESULTS: The volume (P < 0.0001) and the weight (P < 0.0001) of the saliva were increased by using a 980 nm diode laser. While, in smokers and diabetic patients with hypertension, no significant difference was observed in either saliva volume (P = 0.593, P = 0.092) or weight (P = 0.785, P = 0.089). CONCLUSION: In this study, the volume and weight of the unstimulated saliva in most patients with xerostomia increased with using PBM.

A New Model of Salivary Pacemaker-A Proof of Concept and First Clinical Use.

Background and Objectives: Saliva is of utmost importance for maintaining oral health. Management of saliva flow rate deficiency recently includes salivary neuro-electrostimulation. The aim of this paper is to present a new model of salivary pacemaker-the MICROSAL device (MD), an intelligent, miniaturized, and implant-supported oral device used for salivary stimulation. Materials and Methods: This report presents the development, calibration, and first clinical tests which involved the MD. The novel features of this device are the pH sensor and the fact that it communicates with the patient's smartphone, where oral wetness and pH are graphically exposed. Saliva samples were taken before and after the MD was used on a 68-year-old patient suffering from post-irradiation xerostomia, and albumin and total protein were analyzed. Results: The device uses up to 3 V and time intervals of 2 s seconds for stimulation. The total volume of all saliva samples collected during the clinical trial was almost seven times higher after the device was used. Albumin decreased from a maximum of 0.15 g/dL to 0.04 g/dL, and total proteins from 0.65 g/dL to 0.21 g/dL, after salivary stimulation. Conclusions: The MD increased saliva secretion of the patient, and we are confident it will be a good solution for future management of salivary gland hypofunction.

Mental Rehearsal Improves Passing Skill and Stress Resilience in Rugby Players.

PURPOSE: Mental rehearsal is commonly employed, with positive visualization proposed to enhance complex skill performance. Additionally, video stimulus has been associated with enhanced kinesthetic sensations and rapid hormone fluctuations that may contribute to enhancing mental rehearsal and the conscious and unconscious emotional state for skill execution. Here, we assessed the impact of a 15-minute mental rehearsal intervention on rugby-specific tasks and the associated hormone profile. METHODS: Professional rugby players (N = 10) volunteered for a randomized crossover study. They completed three 15-minute preparatory phases (positive or negative video-guided mental rehearsal or self-directed mental rehearsal alone) prior to an exercise stressor and rugby-specific passing task. Salivary testosterone and cortisol were monitored to assess stress responses. RESULTS: Performance during the rugby passing task was improved following the positive video condition (91% [7.4%]) compared to the negative video (79% [6.0%]; ES: 1.22 ± 0.75) and self-visualization (86% [5.8%]; ES: 0.58 ± 0.75), with a significant correlation observed between passing performance and salivary testosterone (r = .47 ± .34, P = .0087). Positive video imagery prior to an exercise stressor also significantly enhanced physiological stress resilience (r = .39 ± .36, P = .0352). CONCLUSIONS: This pilot study demonstrates that mental rehearsal was enhanced by appropriate, context-specific video presentation. We propose that the interaction between sex steroids, the adrenal axis, and subsequent conscious and unconscious behaviors may be relevant to competitive rugby. Specifically, we suggest that relatively elevated free testosterone imparts a degree of stress resilience, which may lead to enhanced expression of competitive behaviors and provide an enhanced state for rugby skill execution.

A capacitive laser-induced graphene based aptasensor for SARS-CoV-2 detection in human saliva.

SARS-CoV-2 virus induced CoVID-19 pandemic has accelerated the development of diagnostic tools. Devices integrated with electrochemical biosensors may be an interesting alternative to respond to the high demand for testing, particularly in contexts where access to standard detection technologies is lacking. Aptamers as recognition elements are useful due to their stability, specificity, and sensitivity to binding target molecules. We have developed a non-invasive electrochemical aptamer-based biosensor targeting SARS-CoV-2 in human saliva. The aptamer is expected to detect the Spike protein of SARS-CoV-2 wildtype and its variants. Laser-induced graphene (LIG) electrodes coated with platinum nanoparticles were biofunctionalized with a biotin-tagged aptamer. Electrochemical Impedance Spectroscopy (EIS) for BA.1 sensing was conducted in sodium chloride/sodium bicarbonate solution supplemented with pooled saliva. To estimate sensing performance, the aptasensor was tested with contrived samples of UV-attenuated virions from 10 to 10,000 copies/ml. Selectivity was assessed by exposing the aptasensor to non-targeted viruses (hCoV-OC43, Influenza A, and RSV-A). EIS data outputs were further used to select a suitable response variable and cutoff frequency. Capacitance increases in response to the gradual loading of the attenuated BA.1. The aptasensor was sensitive and specific for BA.1 at a lower viral load (10-100 copies/ml) and was capable of discriminating between negative and positive contrived samples (with strain specificity against other viruses: OC43, Influenza A, and RSV-A). The aptasensor detected SARS-CoV-2 with an estimated LOD of 1790 copies/ml in contrived samples. In human clinical samples, the aptasensor presents an accuracy of 72%, with 75% of positive percent of agreement and 67% of negative percent of agreement. Our results show that the aptasensor is a promising candidate to detect SARS-CoV-2 during early stages of infection when virion concentrations are low, which may be useful for preventing the asymptomatic spread of CoVID-19.

Effects of the salivary gland massage program in older type 2 diabetes patients on the salivary flow rate, xerostomia, swallowing and oral hygiene: A randomized controlled trial.

AIM: To evaluate the effect of a salivary gland massage program to improve salivary flow, swallowing, and oral hygiene in older type 2 diabetes patients. METHODS: This randomized control trial consisted of 73 older diabetes patients with a low salivary flow; 39 and 34 in intervention and control groups, respectively. The intervention group received a salivary gland massage from a trained dental nurse, whereas the control group received a dental education. The salivary flow rates were collected using spit methods at baseline, 1-month and 3-month follow up. All participants were examined for the objective and subjective symptoms of xerostomia, the Simplified Debris Index and the Repetitive Saliva Swallowing Test. RESULTS: After 3 months, the resting (0.32 vs 0.14 mL/min, P < 0.001) and stimulating salivary flow (3.66 vs 2.83 mL/min, P = 0.025) in the intervention group was significantly higher than the control group. The objective symptoms in the intervention group were significantly lower than the control group after 3 months (1.41 vs 2.26, P = 0.001). The participants who were able to swallow least three times in the Repetitive Saliva Swallowing Test in the intervention group increased by 35.89% after 3 months, whereas the control group increased by 8.82%. Oral hygiene was improved in both groups, but the changes in the intervention were significantly greater than in the control group. CONCLUSIONS: The 3-month salivary glands massage program increases the salivary flow rate, and affects swallowing, objective dry mouth symptoms and oral hygiene in older patients with type 2 diabetes. Geriatr Gerontol Int 2023; 23: 549-557.

Impact of Dietary Nitrate on the Recovery of Therapy-related Vascular Health Impairments Following Standard Periodontal Aftercare Therapy: a Hypothesis-generating Subanalysis.

This follow-up study assessed the impact of a nitrate-rich diet on salivary nitrate/nitrite levels and the recovery of therapy-induced vascular impairments in a cohort of 39 periodontitis patients treated by standard subgingival mechanical plaque removal (PMPR). At baseline, saliva samples for nitrate/nitrite analysis were collected, and peripheral/central blood and augmentation pressure was documented using the Arteriograph recording system. Immediately after, PMPR vascular parameters were reassessed. All study patients received a randomly allocated supply of a lettuce beverage to be consumed for 14 days, containing either a daily dosage of 200 mg nitrate (test group, n = 20) or being void of nitrate (placebo group, n = 19). At day 14, salivary and vascular parameters were reassessed. Initial salivary and vascular parameters did not differ significantly between the groups. PMPR impaired all vascular parameters in both groups with no differences between the groups. At day 14, salivary nitrate/nitrite levels of the test group were significantly elevated compared to baseline. All vascular parameters had significantly recovered from the impairment inflicted by PMPR. In the placebo group, by contrast, salivary parameters did not differ significantly from baseline, and the recovery of impaired vascular parameters was restricted to a significant improvement of diastolic blood pressure. Correlation analysis identified a significant inverse correlation between salivary nitrate/nitrite sum and central/peripheral blood pressure and augmentation pressure. In conclusion, the data of this subanalysis suggest that increasing salivary nitrate/nitrite levels by a diet rich in nitrate may improve recovery of therapy-induced vascular impairments after PMPR.

Assessing stress using repeated saliva concentration of steroid hormones in dementia care dyads: results from a controlled pilot care music intervention.

Background: Stress-related biomarkers have the potential to provide objective measures of whether interventions directed at people with dementia (PWD) and their family caregivers (FCG) are successful. The use of such biomarkers has been limited by logistical barriers to sample collection. Objective: Explore saliva concentration of steroid hormones in dementia care dyads during a music intervention. Methods: Consecutive PWD attending a memory evaluation center and their FCG were allocated to either an intervention-with-music or a non-intervention control group. All were living at home. Stress biomarkers, salivary cortisol and dehydroepiandrosterone sulfate (DHEA-S) samples were collected by the PWD and their FCG, in the morning and evening, 5 days a week, for 8 consecutive weeks. Biomarker concentrations of the intervention and the control groups were compared at week 8, in an intention-to-treat approach with adjustment for baseline value. Results: Twenty-four PWD in the intervention group and 10 in the control group, and their FCG were included in the analyses. The mean number of morning saliva collections was similar in the intervention and the control groups, ranging from 4.3 to 4.9 per participant weekly during the first 7 weeks, declining to 3.3 during week 8. Median log morning cortisol (pg/mL) among caregivers was lower in the intervention group than in the control group (8.09 vs. 8.57, P = 0.0133). Conclusion: This study demonstrates that music intervention was associated with lower morning saliva cortisol concentrations for FCGs.