Stress/Immune Biomarkers in Saliva among Children with ADHD Status.

Background. This cross-sectional study aimed to evaluate stress and immune biomarkers in saliva samples of attention-deficit/hyperactivity disorder ADHD compared to healthy non-ADHD children. Material and methods. A total of 132 children under 11 years old (8.5 ± 1.1) enrolled in a cross-sectional study: with confirmed ADHD (n = 60) and healthy controls (n = 72). The clinical evaluation included physical measurements (height, waist, hip circumference, body weight, body mass index BMI, BMI z-score) and unstimulated saliva collection and measurements of free cortisol, salivary alpha-amylase (sAA), and secreted immunoglobulins (sIgA, IgG, and IgM) with quantitative assay (ELISA) analysis. Unpaired t-test, Welch test, or Mann-Whitney U test were applied for group comparisons when appropriate, and the correlation between variables was analyzed with Spearman's rank coefficient. Results were considered significant at p < 0.05. Results. In the ADHD group, body weight (p ≤ 0.01), BMI (p ≤ 0.009), and hip circumference (p ≤ 0.001) significantly differed, while waist size and BMI z-score did not (p > 0.05). Significant elevation of the salivary sAA (p = 0.03), sIgA (p = 0.02), and IgM (p ≤ 0.001) biomarkers were detected, without differences in the morning cortisol (p > 0.05). Significant correlations between cortisol and BMI, hip size, and IgA, as well as between IgG and sAA and IgA were obtained. Conclusions. Saliva can be used to monitor ADHD status with regard to biomarkers indicating the hypothalamus-pituitary-adrenal axis, as HPA axis, and sympathetic activity. The results indicate that morning collection of saliva in contrast to unchanged salivary cortisol, may evaluate mentioned above system dysregulations by measurements of sAA and immunoglobulins among ADHD children.

Impact of a toothpaste with microcrystalline hydroxyapatite on the occurrence of early childhood caries: a 1-year randomized clinical trial.

The aim of this trial was to determine whether a toothpaste with microcrystalline hydroxyapatite is not inferior to a fluoride toothpaste in prevention of caries in children. This double-blinded randomized control trial compared two toothpastes regarding the occurrence of caries lesions using International Caries Detection and Assessment System (ICDAS) ≥ code 1 on the primary dentition within 336 days. The test group used a fluoride-free hydroxyapatite toothpaste three times daily while control group used a toothpaste with fluoride. 207 children were included in the intention-to-treat analysis; 177 of them finished the study per protocol. An increase in caries ICDAS ≥ code 1 per tooth was observed in 72.7% of the hydroxyapatite-group (n = 88), compared with 74.2% of the fluoride-group (n = 89). The exact one-sided upper 95% confidence limit for the difference in proportion of participants with ICDAS increase ≥ 1 (-1.4%) was 9.8%, which is below the non-inferiority margin of 20% demonstrating non-inferiority of hydroxyapatite compared to the fluoride control toothpaste. This RCT showed for the first time, that in children, the impact of the daily use of a toothpaste with microcrystalline hydroxyapatite on enamel caries progression in the primary dentition is not inferior to a fluoride control toothpaste (Clinical Trials NCT03553966).

Salivary Biomarkers (Opiorphin, Cortisol, Amylase, and IgA) Related to Age, Sex, and Stress Perception in a Prospective Cohort of Healthy Schoolchildren.

BACKGROUND: The use of easily accessible biomarkers for assessing young patients' health is weighty. This cohort study is aimed at measuring stress/immune biomarkers in the saliva of healthy school-age children and comparing subgroups according to age, sex, and stress perception. Material and Methods. 503 children under 12 years old (8.7 ± 1.3) were included with anthropometric evaluation (height, waist, hip circumference, body weight, and body mass index (BMI)). Levels of opiorphin (OPI), free cortisol, alpha-amylase (sAA), and secreted immunoglobulin (sIgA) were determined by quantitative assays (ELISA) in unstimulated saliva. Unpaired t-test, Welch test, and Mann-Whitney U test were applied for appropriate group comparisons, and the correlation between variables was analyzed with Spearman's rank coefficient. Results were considered significant at p < 0.05. RESULTS: sIgA and sAA exhibited significant differences depending on age and sex: IgA (ng/mL): 86 ± 68.6 vs. 104.9 ± 72.1 for (6-7 y.o.) and (8-11 y.o.), respectively, and 108.1 ± 80.1 vs. 94.6 ± 62.2 for male and females, respectively; sAA (U/mL): 78.9 ± 54.4 vs. 100.5 ± 81.2 for (6-7 y.o.) and (8-11 y.o.). No difference related to age or sex between groups was observed for cortisol and OPI. However, OPI levels were higher and correlated to prior stress exposure in children (0.31 ± 0.4 vs. 0.26 ± 0.5 ng/mL, p = 0.031). sAA was negatively correlated to low mood self-declaration in children in the last two weeks (r = -0.10, p = 0.045). CONCLUSIONS: sIgA and sAA can be used as sex- and age-related biomarkers in children 6-12 y.o., which is not the case for free cortisol and opiorphin. However, OPI reflected previous exposure to stress, suggesting its use for evaluating stress-related changes in children.

Is there a link between stress and immune biomarkers and salivary opiorphin in patients with a restrictive-type of anorexia nervosa?

Objectives: Opiorphin is a salivary peptide with analgesic and antidepressant properties. Its relationship with the hypothalamic-pituitary-adrenal, autonomic nervous and immune systems may provide understanding of chronic stress, especially in anorexia nervosa (AN). This study investigated a possible correlation between opiorphin and stress/immune biomarkers, cortisol, salivary alpha-amylase (sAA) and secretory immunoglobulin A (sIgA), in saliva of patients with restrictive-type AN.Methods: A case-control clinical trial was conducted in 92 AN patients (+75 healthy controls). Unstimulated salivary samples were taken during the acute stage of AN, measurements of cortisol, sAA, sIgA and opiorphin were performed with a quantitative assay (enzyme-linked immunosorbent assay, P < 0.05).Results: AN patients displayed an increase in cortisol (P < 0.001) and sIgA (P < 0.001) but not in sAA (P = 0.279) levels. Distinct correlation between these two parameters and body-weight indexes were observed. Opiorphin levels were neither correlated to stress and immune biomarkers, nor to salivary flow rate.Conclusions: The effect of stress responses can be reliably assessed in saliva in AN patients. The difference between sIgA and cortisol indicate that they can both be used for mental stress assessment in saliva. Modulation of opiorphin by chronic stress was not confirmed. Unchanged sAA indicates a partial adaptation of human organism to severe condition during malnutrition.

Salivary opiorphin levels in anorexia nervosa: A case-control study.

Objectives: Opiorphin is a physiological inhibitor of peptidases inactivating endogenous opioids displaying strong analgesic properties without undesirable side effects, antidepressant properties or hormonal dependency. It might therefore play an important role in patients with painful diseases related to neuro-hormonal dysregulation of the nervous system, affecting saliva secretion and composition such as anorexia nervosa (AN). The main objective aim of this study was to compare the level of opiorphin in saliva of patients with AN to matched subjects free of eating disorders.Methods: A case-control clinical trial was conducted in 68 AN patients and 43 healthy matched control subjects. Depression symptoms were assessed with the self-scored questionnaire Beck Depression Inventory (BDI) and salivary samples were taken during the acute stage of AN (BMI <15 kg/m2) for measuring opiorphin. Opiorphin levels were measured with a quantitative assay using a commercial immunoenzymatic Elisa kit (cat no. EH1927, Wuchan, Hubei, China).Results: No statistically significant difference was found in salivary opiorphin levels between the AN and control groups, (P = 0.499, Mann-Whitney U-test). Positive correlations to duration of the disease, BDI and bodyweight in AN patients were evidenced.Conclusions: Measurement of salivary opiorphin levels cannot be used as a marker of AN but may allow new perspectives in monitoring AN in its early stages.