Salivary Cortisol Determination Research Articles

Blood and Salivary Cortisol Variations in Athletes in Relation to Cardiopulmonary Exercise Testing.

Background and Objectives: Cortisol is a valuable marker for assessing the body's response to any form of stress. We conducted this study in order to evaluate the variations of salivary and serum cortisol levels in professional football players in relation to cardiopulmonary exercise testing (CPET) and their significance in potentially evaluating overtraining in athletes. Also, the question of whether salivary cortisol determination could fully substitute serum sampling was addressed. Materials and Methods: A total of 19 male professional football players were evaluated by measuring serum cortisol levels at rest (T0) and immediately after a CPET (T1) and salivary cortisol levels at rest (T0), 10 min after a CPET (T2), and 30 min after a CPET (T3). Results: T0 serum cortisol showed a statistically significant correlation with the oxygen uptake at the anaerobic threshold divided by the body weight (VO2-AT/weight), as did the T2 salivary cortisol with the maximum oxygen uptake at the anaerobic threshold (VO2-AT) and VO2-AT/weight. T0 salivary cortisol was significantly correlated with the subjects' height and the predicted O2 pulse. Conclusions: While some correlations were discovered, they are insufficient to recommend cortisol as a routine biomarker in athletes' evaluation. However, significant correlations were established between salivary and serum determinations, meaning that the non-invasive procedure could substitute venous blood sampling.

Determination of salivary cortisol and salivary pH level in gaming teenagers - A cross-sectional study.

Internet gaming disorder and cybervictimization has a disproportionate increase during the COVID 19 pandemic. Teenagers and adolescents have high risk for addiction to online gaming. Online gaming increases the stress level of the users affecting the general and the oral health. The study aims to determine the salivary cortisol and salivary pH among the gaming teenagers. The cross-sectional study recruited 45 gaming teenagers with the mean age of 15.5 years. The participants were stratified according to the type of online gaming played as mind, aggressive and betting gaming. Their salivary pH and salivary cortisol were assessed. The collected data was analyzed using SPSS version 23. The mean salivary pH was found to be less among females playing aggressive games (7.11±0.30) and males playing mind games (7.24±0.16). Salivary cortisol level was found to be statistically high in males (6.66±1.33) and females (7.82±0.66) playing aggressive games. The sub-group analysis of salivary pH and cortisol for gender showed a significant difference in the mean salivary cortisol level (p<0.05) with no difference in the salivary pH (p>0.05). Based on the number of hours spent in online gaming, there was a significant increase in the mean salivary cortisol level among the participants playing >3h (p<0.05). Salivary cortisol is high among individuals playing aggressive games compared to mind and betting games. Also, salivary cortisol is high in females compared to males with no change in salivary pH.

Preference for and maintenance of interest in suspended enrichment toys in confined growing pigs

The objective of the present study was to evaluate the preference for, interest in, and behavior of growing pigs provided with three different suspended enrichment toys in a four-compartment environmental preference chamber (EPC). Sixty pigs at 70±2 days of age were divided into two contemporary groups. There were two distinct areas in each compartment of the EPC: one isolated area where two control pigs were housed with access to one type of toy, and one area with free access to the other compartments where five test pigs were housed for 5 days (D-1, D0, D1, D2, D3). Each week, a new group of pigs was introduced into the EPC; this procedure was repeated four times with a different contemporary group. Three different suspended toys (PET bottle; sisal rope; chain) were offered from D1 to D3 and were removed and washed at the end of the day. The toys and a control were randomly arranged in each EPC compartment (control; test). Focal animal sampling with recording interval of 10 min was used to determine the occurrence of behaviors (agonistic, idle, eating, drinking, vocalizing, stereotypic, positive, use of toy, others) from 8:00 to 11:00 h and from 13:00 to 16:00 h. Only for test pigs, for the ‘use of toy’, we also recorded which toy was being used as well as the duration it was used for. Samples for the determination of salivary cortisol and respiratory rate were taken on alternate days (D0; D2) at 7:00 and 13:00 h. Data were analyzed by ANOVA with fixed effects of group (test, control), day (D1, D2, D3), and period (7:00, 13:00 h - cortisol), as well as their interaction. Means were compared by the Tukey-Kramer or t-test (P < 0.05). The sisal rope was the preferred toy in the test group (P < 0.05), with a total duration of 70.8±3.9 min and 36.5±3.7 bouts, followed by the PET bottle (14.7±2.2 min; 12.0±1.6 bouts) and the chain (15.8±1.4 min; 12.4±0.9 bouts) (P > 0.05). Test pigs also exhibited fewer idle bouts compared to control pigs (P < 0.05). Control pigs preferred the sisal rope (P < 0.05). Cortisol concentrations did not differ significantly between treatment groups or sampling days, although they numerically decreased by 34.6% (7:00 h) and 17.8% (13:00 h) from D0 to D2. In conclusion, the preferred toy was the sisal rope, with a slight reduction in the number of bouts and duration over three days; however, this toy was still attractive to the pigs and reduced idle behavior when they have the possibility of choosing toys.

Usefulness of salivary cortisol as a marker of secondary adrenal insufficiency in paediatric patients

Background: The main cause of adrenal insufficiency (AI) in paediatric patients is prolonged treatment with corticosteroids. Determination of plasma cortisol (PC) during ACTH test is the most used adrenal function indicator in clinical practice. However, determination of salivary cortisol (SC), a simple test especially useful in children in order to avoid invasive procedures, can be used as an alternative technique for the diagnosis of adrenal disease. Methods: A two-year prospective study (January 2014-January 2016) in paediatric patients (2-18 years of age) treated with corticosteroids for more than fifteen days, who were investigated for suspected AI. Low-dose ACTH test was used to determine adrenal function and samples for SC and PC were obtained simultaneously in basal situation and during the test (at 30, 60 and 90 minutes). Results: 230 samples (118 PC-112 SC) of 30 studies belonging to 20 patients (4 males), mean age 10.93 years ± 3.69 SD. Pearson’s correlation coefficient showed a positive correlation between PC and SC (r = 0.618, p &lt; 0.001). All the studies with some determination of PC higher than 18 μg/dL (n = 8) had a SC peak higher than 0.61 μg/dL with a specificity of 66.67% and a sensitivity of 93.94% (ROC analysis). Conclusion: Measurement of SC is a less invasive, easier and quicker test than PC to measure plasma free cortisol levels. In our study, a SC peak in low-dose ACTH test higher than 0.61 μg/dL was able to discriminate patients without AI, and proved to be a useful tool in the initial evaluation of children with suspected AI.

Can stress predict delivery date?: Role of chronic and acute stress to the threatened preterm labor as predictors of delivery date

IntroductionThreatened preterm labor (TPL) is a traumatic event during pregnancy that involves a threat to the physical integrity of the upcoming baby. Despite biomarkers would be the strongest delivery date predictors, an assessment of chronic and acute stress response to TPL diagnosis may improve this prediction.ObjectivesThe objective is to predict delivery date in women with TPL based on their response to this diagnosis and chronic stressors, along with relevant obstetric variables.MethodsA prospective cohort study was conducted with a sample was formed by 157 pregnant women with TPL diagnosis between 24 and 31 weeks. Determination of salivary cortisol, α-amylase levels, along with anxiety and depression symptoms were measured to estimate stress response to TPL. Cumulative life stressors as traumas, social and familiar functioning were also registered. To examine the effect of the possible predictor variables of delivery date, linear regression models were used.ResultsA correlation was found between the variables of response to chronic stress and between the variables of psychological response to stress. The main predictors of preterm delivery were low family adaptation, higher BMI, higher cortisol levels, and the week of diagnosis of TPL (&lt;29 weeks of gestation).ConclusionsThe best predictor of delivery date was the combination of the stress response to the diagnosis of TPL measured by cortisol in saliva, cumulative life stressors (mainly family adaptation) and obstetric factors (week TPL and BMI). Through psychosocial therapeutic intervention programs, it is possible to influence this modifiable predictive factors of preterm birth in symptomatic women.

Non-enzymatic electrochemical determination of salivary cortisol using ZnO-graphene nanocomposites.

Electrochemically deposited ZnO nanoparticles on a pencil graphite electrode (PGE) coated with graphene generate a noteworthy conductive and selective electrochemical sensing electrode for the estimation of cortisol. Electrochemical techniques such as cyclic voltammetry (CV) analysis and electrochemical impedance spectroscopic (EIS) tests were adopted to analyze and understand the nature of the modified sensor. Surface morphological analysis was done using various spectroscopic and microscopic techniques like X-ray photoelectron spectroscopy (XPS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). Structural characterization was conducted by X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). The effect of scan rate, concentration, and cycle numbers was optimized and reported. Differential pulse voltammetric (DPV) analysis reveals that the linear range for the detection of cortisol is 5 × 10−10M − 115 × 10−10 M with a very low-level limit of detection value (0.15 nM). The demonstrated methodology has been excellently functional for the determination of salivary cortisol non-enzymatically at low-level concentration with enhanced selectivity despite the presence of interfering substances.

Can Haematological and Hormonal Biomarkers Predict Fitness Parameters in Youth Soccer Players? A Pilot Study.

The study aimed to investigate the correlations among immune, haematological, endocrinological markers and fitness parameters, and assess if the physiological parameters could be a predictor of fitness values. Anthropometric, physical evaluations (countermovement jump—CMJ, 10 m sprint, VO2max, repeated sprint ability—RSA total time and index) and determination of blood (IL-6, IL-10, IL-17A and tumour necrosis factor) and salivary (testosterone and cortisol) samples parameters in 28 young male soccer players (age: 13.0 ± 0.2 years, body mass index (BMI): 19.5 ± 2.2 kg/m2) were analysed. To evaluate the dependence of the variables related to athletic performance, multiple linear regression with backward stepwise elimination was considered. A significant regression equation was found in CMJ (F(5,16) = 9.86, p < 0.001, R2 adjusted = 0.679) and in the RSA index (F(5,16) = 15.39, p < 0.001, R2 adjusted = 0.774) considering only five variables, in a 10 m sprint (F(4,17) = 20.25, p < 0.001, R2 adjusted = 0.786) and in the RSA total time (F(4,17) = 15.31, p < 0.001, R2 adjusted = 0.732) considering only four variables and in VO2max (F(9,12) = 32.09, p < 0.001, R2 adjusted = 0.930) considering nine variables. Our study suggests the use of regression equations to predict the fitness values of youth soccer players by blood and saliva samples, during different phases of the season, short periods of match congestion or recovery from an injury.

Salivary cortisol measurement in horses: immunoassay or LC-MS/MS?

The aim of the present study was to measure salivary cortisol concentrations of horses before and after hypothalamic-pituitary-adrenal (HPA) axis stimulation by means of liquid chromatography-tandem-mass spectrometry (LC-MS/MS) and an immunoassay (cELISA) for method comparison. Nine clinically healthy horses participated in the study. An ACTH stimulation test was performed. Saliva samples were collected before (T0) and 60 (T60) min after intravenous injection of 1 μg/kg BW synthetic ACTH1-24. LC-MS/MS was assessed for the determination of equine salivary cortisol. The results of these measurements were then compared to the results obtained by a cELISA, which has previously been validated for use in horses. The Pearson correlation coefficient was calculated and showed no correlation at T0 (r = −0.2452; P = 0.5249) and significantly correlated results at T60 (r = 0.8334; P = 0.0053). Bland-Altman-Plots of T60 revealed that immunoassay measurements led to higher outcome values than LC-MS/MS. On average, immunoassay results were 2.3 times higher. Poor agreement between both methods at T0 is potentially a consequence of inaccuracy in the very low measuring range of the immunoassay, and to a smaller extent, structurally similar cross-reacting agents and matrix effects, which might bias the results. Overestimation of immunoassay results at T60 might be due to different standardization of both methods, non-avoidable matrix effects on the antigen-antibody interaction in the ELISA, and possibly cross-reactions of other steroids. While immunoassay measurements of equine salivary cortisol yielded higher but reasonably correlated results for elevated cortisol concentrations after stimulation of the HPA axis, LC-MS/MS provided more accurate results, particularly for baseline cortisol concentrations close to the limit of detection of the ELISA.

Determination of salivary cortisol to assess time-related changes of the adrenal response to stress in critically ill patients

BackgroundThe value of salivary cortisol measurement to study stress-related adrenal response is controversial. The study aim was to assess the role of salivary cortisol measurement to detect time-related changes of adrenal response in critically ill patients. Patients and methodsPatients with organ failure, sepsis or trauma were prospectively recruited in the Emergency Department. Serum and salivary cortisol were measured at baseline (T0) and after 48 h (T48). In 33 patients ACTH test was also done. ResultsFifty-five patients were studied and classified as septic (22) or non-septic (33). We found a significant correlation between serum and salivary cortisol at T0 and T48. No patient had baseline serum cortisol < 276 nmol/L and salivary cortisol significantly decreased at T48 in almost all patients. A delta serum cortisol < 250 nmol/L after ACTH was found in only 4 patients who showed elevated baseline cortisol levels. ConclusionWe found that reduced baseline and post-ACTH cortisol levels are uncommon in our samples. In patients able to provide adequate saliva samples, salivary cortisol may be used to check the degree of stress-induced response and appears as a suitable tool for multiple measurements over time.

Salivary Cortisol Determination in ACTH Stimulation Test to Diagnose Adrenal Insufficiency in Patients with Liver Cirrhosis.

Purpose The prevalence of adrenal insufficiency (AI) in patients with decompensated liver cirrhosis is unknown. Because these patients have lower levels of cortisol-binding carrier proteins, their total serum cortisol (TSC) correlates poorly with free serum cortisol (FC). Salivary cortisol (SaC) correlates better with FC. We aimed to establish SaC thresholds for AI for the 250 μg intravenous ACTH test and to estimate the prevalence of AI in noncritically ill cirrhotic patients. Methods We included 39 patients with decompensated cirrhosis, 39 patients with known AI, and 45 healthy volunteers. After subjects fasted ≥8 hours, serum and saliva samples were collected for determinations of TSC and SaC at baseline 0'(T0) and at 30-minute intervals after intravenous administration of 250 μg ACTH [30'(T30), 60'(T60), and 90'(T90)]. Results Based on the findings in healthy subjects and patients with known AI, we defined AI in cirrhotic patients as SaC-T0< 0.08 μg/dL (2.2 nmol/L), SaC-T60 < 1.43 μg/dl (39.5 nmol/L), or ΔSaC<1 μg/dl (27.6 nmol/L). We compared AI determination in cirrhotic patients with the ACTH test using these SaC thresholds versus established TSC thresholds (TSC-T0< 9 μg/dl [248 nmol/L], TSC-T60 < 18 μg/dl [497 nmol/L], or ΔTSC<9 μg/dl [248 nmol/L]). SaC correlated well with TSC. The prevalence of AI in cirrhotic patients was higher when determined by TSC (48.7%) than by SaC (30.8%); however, this difference did not reach statistical significance. AI was associated with sex, cirrhosis etiology, and Child-Pugh classification. Conclusions Measuring SaC was more accurate than TSC in the ACTH stimulation test. Measuring TSC overestimated the prevalence of AI in noncritically ill cirrhotic patients.

Is EIPH a misnomer and could it be hampering our attempts to understand the etiology of the condition?

Salivary cortisol provides information about free plasma cortisol concentration, and salivary sampling is a noninvasive well-tolerated procedure. The aim of this study was to validate a commercial enzyme immunoassay for the determination of salivary cortisol in donkeys. Saliva samples were collected in four donkey stallions on 13 nonconsecutive days at 8:30 AM to avoid circadian variation. Animals were already accustomed to be handled. Saliva was collected using a swab inserted at the angle of the lips, placed onto the tongue for 1 minute and returned into a polypropylene tube. Tubes were centrifuged, and at least 1 mL of saliva was aspirated from each sample and frozen at −20°C until analysis. A commercial enzyme immunoassay kit without extraction was used for determination of cortisol in saliva. Median cortisol concentrations with minimum and maximum value were calculated. Recovery of cortisol standard in donkey saliva was between 97.3% and 99.7%, and serial dilution of donkey saliva samples with assay buffer resulted in changes in optical density parallel to the standard curve. Cross-reactivity of the antiserum was 10.4% with 11-deoxycortisol, 5.2% with corticosterone, 0.4% with 11-deoxycorticosterone, 0.2% with cortisone, and <0.1% with testosterone, progesterone, and estradiol. The intra-assay coefficient of variation was 10.7%, the interassay variation was 8.0%, and the minimal detectable concentration was 0.01 ng/mL. The results of the present study demonstrate the validity of a commercial kit to determine the concentration of cortisol in donkey saliva as already reported in other species.

Determination of Salivary Cortisol in Donkey Stallions

Salivary cortisol provides information about free plasma cortisol concentration, and salivary sampling is a noninvasive well-tolerated procedure. The aim of this study was to validate a commercial enzyme immunoassay for the determination of salivary cortisol in donkeys. Saliva samples were collected in four donkey stallions on 13 nonconsecutive days at 8:30 AM to avoid circadian variation. Animals were already accustomed to be handled. Saliva was collected using a swab inserted at the angle of the lips, placed onto the tongue for 1 minute and returned into a polypropylene tube. Tubes were centrifuged, and at least 1 mL of saliva was aspirated from each sample and frozen at −20°C until analysis. A commercial enzyme immunoassay kit without extraction was used for determination of cortisol in saliva. Median cortisol concentrations with minimum and maximum value were calculated. Recovery of cortisol standard in donkey saliva was between 97.3% and 99.7%, and serial dilution of donkey saliva samples with assay buffer resulted in changes in optical density parallel to the standard curve. Cross-reactivity of the antiserum was 10.4% with 11-deoxycortisol, 5.2% with corticosterone, 0.4% with 11-deoxycorticosterone, 0.2% with cortisone, and <0.1% with testosterone, progesterone, and estradiol. The intra-assay coefficient of variation was 10.7%, the interassay variation was 8.0%, and the minimal detectable concentration was 0.01 ng/mL. The results of the present study demonstrate the validity of a commercial kit to determine the concentration of cortisol in donkey saliva as already reported in other species.

Evaluation of the Performance of an Automated Chemiluminescent Method (Access Beckman Coulter) for Determination of Salivary Cortisol and Clinical Utility

The diagnostic procedure for the evaluation of Cushing’s syndrome is performed by any of the following biochemical tests: urine free cortisol, salivary cortisol at 11 pm and serum cortisol post 1 mg of dexamethasone. Collection of saliva samples is simple and noninvasive, thus being a method of choice for the evaluation of risk populations. The aim of this work is to analyze the performance of an automated chemiluminescent method for measurement of salivary cortisol at 11 pm according to the new quality guidelines and assess its clinical utility. Cortisol levels were measured in samples obtained by passive drooling from 32 healthy subjects and 9 patients with Cushing’s syndrome. Matrix effect, linearity, limit of blank, limit of quantitation, recovery and diagnostic performance were assessed. The Unicel 600 DXI Access Beckman Coulter chemiluminescent automated analyzer was used. The standard curve provided by the manufacturer was adapted to measure cortisol concentrations in saliva. Matrix effect: equation of the curve using salivary matrix: y=-1.824x+3.491 (95% CI=-2.068 to -1.582) vs. Equation of the curve using diluent matrix: y=-1.833x+3.394 (95% CI=-1.961 to -1.704). There is overlapping of both curves. Linearity: linear assay between 1.8 nmol/L and 108.0 nmol/L. Limit of blank: 0.1 nmol/L. Limit of quantitation: 1.8 nmol/L (TAE of 25%). Recovery: standard cortisol solution concentration 5 nmol/L: 102%; 10 nmol/L: 107%; 40nmol/L: 115%. Diagnostic performance: median and ranges in healthy subjects: 2.0 nmol/L (<2.0-9.0 nmol/L); Cushing’s syndrome: 30.3 nmol/L (15.4-61.0 nmol/L). ROC curve cutoff value: 9.0 nmol/L (100% Specificity; 100% Sensitivity; AUC=1.00). The method used provides excellent analytical performance for cortisol measurement in saliva at 11 pm, which makes it a valuable biochemical tool both for screening populations at risk for Cushing’s syndrome and for the follow-up and diagnosis of this condition.

Validation model of a laboratory-developed method for the ISO15189 accreditation: The example of salivary cortisol determination

BackgroundThe ISO 15189:2012 describes the requirements for quality and competence, which are specific for all sectors of a medical laboratory. Laboratory-developed methods, standard methods used outside their intended scope and validated methods subsequently modified shall be validated. The main objective of validation of an examination procedure (EP) is to demonstrate its fitness-for-purpose. The aim of the present study is to illustrate a model to validate laboratory-developed methods in agreement with the ISO 15189:2012 requirement, through the example of salivary cortisol (sF) measurement by liquid chromatography tandem mass spectrometry (LC-MS/MS). MethodsThe proposed validation model for a laboratory developed method includes 6 steps: 1) analysis of available scientific documentation pertinent to specific EP; 2) evaluation of EP intended use; 3) identification of performance characteristics to evaluate; 4) definition of experimental procedure; 5) identification of acceptability criterion for results evaluation; 6) preparation of a validation plan; 7) production of a validation certificate. ResultssF is an EP sampled during the accreditation audit and resulted in compliance with the Standard. A structured procedure to certify that the achieved performance specifications are appropriate for the purpose of the test is therefore necessary. ConclusionsA high competence is required for laboratory professionals in order to guarantee that the validated quality specifications have a positive impact on patients' management. The ISO 15189 accreditation is an important tool that allows demonstrating the value of a medical laboratory in the clinical context.

Salivary cortisol and cortisone: UPLC-MS/MS method validation and temporal variability over one week

Aims: The present study aims to provide a comprehensive analytical and biological validation of an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for analysis of salivary cortisol and its inactive metabolite cortisone. Variation in cortisol awakening response (CAR) over one week were investigated. Methods: Saliva samples were collected from 19 healthy volunteers. To determine CAR, participants collected saliva samples at three time points: immediately after awakening, 15 and 30 minutes thereafter. The same procedure was repeated each morning over one week period. In addition, all participants filled in diaries containing information about duration of sleep, time of awakening, smoking, and coffee and alcohol intake. Upon collection, the saliva samples were stored at -20°C until analysis. Prior to analysis, the saliva samples were thawed and spiked with internal standard and extracted using solid phase extraction columns (Oasis Prime-HLB). The identification and quantification of cortisol and cortisone were performed using the developed UPLC-MS/MS method, on a Waters Aquity TQ-XS system. Results: The obtained limits of quantification (LoQ) were 1ng/ml for cortisol and 500pg/ml for cortisone. Intra-assay accuracy values of calibration points were between 83-111%. The mean levels of cortisol and cortisone in the total sample were 3.55 ± 1.99 ng/ml and 10.51 ± 3.46 ng/ml, respectively. In the first 30 minutes after awakening, there was a 70% increase in the average cortisol levels (CAR) and a 49% increase in the levels of cortisone. A high intra-individual variability of CAR was observed over the week (CV ranged between 17.9-68.9%), whereas the inter-individual variability of the average CAR equaled 28.2%. Furthermore, the changes in CAR were related to variables from participants’ diaries. Conclusion: The UPLC-MS/MS method has shown to be a sensitive and specific technique for determination of salivary cortisol and cortisone. However, in the clinical context, CAR data should be interpreted with precaution due to high inter- and intra-individual variability.

Methods for the Quantification of Salivary Cortisol and of a-amylase in Biosensors and Portable Devices

There is an increasing interest in the analysis of salivary biomarkers for medical practice. The objective of this article was to identify the specificity and sensitivity of quantification methods used in biosensors or portable devices for the determination of salivary cortisol and salivary a-amylase. There are no biosensors and portable devices for salivary amylase and cortisol that are used on a large scale in clinical studies. These devices would be useful in assessing more real-time psychological research in the future.

Pinetonina&amp;lt;sup&amp;gt;TM&amp;lt;/sup&amp;gt;, an Intranasally Administered Essential Oil Preparation, Is Effective in Decrease of Cortisol Levels and on the Glutamate Release Modulation

Background: Most anxiety complaints are treated with pharmacological measures involving barbiturates and benzodiazepines, in which they may end up causing tolerance and pharmacological dependence. Integrative approaches such as aromatherapy are used in addition to medications to improve sleep quality and reduce anxiety. Thus, PinetoninTM, a phytocomplex obtained from a blend of essential oils aims to aid in the symptoms of stress and anxiety. Methods: The cytotoxicity of PinetoninTM was evaluated MTT assay using fibroblasts and astrocytes showed reduction in the cell viability only at high concentrations. Evaluation of intracellular calcium and determination of residual glutamate in the supernatant of astrocyte cultures showed agonist action of dihydroxyphenylglycine (DHPG) increasing linearly the concentration of intracellular calcium and the glutamate levels in the supernatants of the cultures. On the other hand, cultures of astrocytes treated with PinetoninTM showed residual glutamate levels in the supernatants reducing proportionally, as well as, intracellular calcium reduction. The determination of salivary cortisol showed a significant reduction in salivary cortisol levels in the group that received PinetoninTM. The evaluation of the electroencephalogram in patients treated with PinetoninTM had a significant increase (P TM treated fibroblasts with control culture. When PinetoninTM and linole are administered in astrocytic cells, there was a reduction of the intracellular concentration of Ca2+ against a control group treated with DHPG agonist. The evaluation of salivary cortisol demonstrated a reduction when the patient group was treated with PinetoninTM by purchasing the results against the group of patients treated with saline. Reinforcing the relaxed state of that group, alpha waves have been increased and reductions in beta waves. Conclusion: The results obtained after intranasal administration of PinetoninTM suggest that this phytocomplex can help reduce the symptoms of stress and the better quality of life.

Determination of salivary α-amylase and cortisol in psoriatic subjects undergoing the Trier Social Stress Test

The study was aimed to investigate the response in psoriatic patients of the two primary neuroendocrine systems, namely the Autonomic Nervous System and the Hypothalamic-Pituitary-Adrenal (HPA) axis, whose main biomarkers of activation are salivary α-amylase and cortisol, respectively. Reliable analytical procedures for the determination of salivary α-amylase enzymatic activity and cortisol concentration in oral fluid were developed. The non-invasive sampling that does not require trained personnel represents one of the main advantages of oral fluid analysis compared to the analysis of blood and its derivatives (e.g. plasma). Since oral fluid offers the advantage of a simple and stress free sample collection, it is particularly useful to quantify stress related molecules.α-amylase activity was measured by a spectrometric method, whereas salivary cortisol by Reversed-Phase High-Performance Liquid Chromatography coupled to Electrospray Ionization Quadrupole Time-of-Flight Mass Spectrometry. The effect of salivary pH and flow rate on salivary α-amylase activity and cortisol were also investigated.Finally, both methods were preliminary tested on oral fluid samples of psoriatic volunteers underwent to a psychosocial stress task (the Trier Social Stress Test), as a first step of a more comprehensive study on the responsiveness of psoriatic subjects to stressors.

Effect of Heat Exposure and Simulated Physical Firefighting Work on Acute Inflammatory and Cortisol Responses.

To examine firefighters' hormonal and immune markers during consecutive days of physical firefighting work performed in hot compared to mild ambient temperatures. Firefighters completed 3 days of simulated physical firefighting work in either hot (HOT condition; n = 19; 33°C) or mild temperature conditions (CON condition; n = 18; 19°C). Participants provided regular daily samples for the determination of salivary cortisol and plasma cytokine levels (IL-6, IL-8, IL-1β, TNF-α, IL-4, and IL-10). The HOT condition elicited higher IL-4 and a trend towards elevated afternoon and evening cortisol when compared to the CON trial. The HOT condition also produced lower levels of IL-1β compared to the CON across time points and a decrease in IL-1β between days of work. IL-6 increased across time points and between work days, but this finding was not different between mild and hot conditions. Immune-endocrine interactions revealed a rise in morning IL-6 that was related to elevated daily cortisol levels, independent of condition. Findings demonstrate the possibility that firefighters are able to regulate normal acute immune and hormonal responses to multiple days of simulated physical work in hot and mild ambient temperatures. Further research is necessary to determine if the responses continue under non-simulated conditions and in response to more extreme temperatures possible on the fire-ground.

Correlation of Plasma and Salivary Cortisol in Extremely Premature Infants

Background: In the neonatal period, cortisol concentrations are expected to increase during significant stress and are important for survival. Extreme preterm infants may develop adrenal insufficiency in the early neonatal period. Cortisol is 90% bound to cortisol binding globulins (CBG) in the circulation; therefore measurements of plasma cortisol can be compromised by conditions that alter CBG levels. Measurement of free cortisol is the best indicator of adrenal glucocorticoid secretion and can be determined in the saliva. Few studies have been reported on salivary cortisol determination in the neonatal period and particularly, extremely premature infants. Methods: There were 65 infants (36 males). Mean gestation 25.3 ± 1.3 weeks. We obtained early morning plasma and salivary cortisol sampling before day 5 of postnatal age from extremely premature babies less than 28 weeks gestation. Saliva was obtained using 4 standard universal swabs by placing one swab at a time in the infant's mouth for 1-2 min. No salivary stimulants were used. Salivary cortisol was measured by competitive ELISA using a commercially available kit SLV-2930 (DRG, Germany) according to the manufacturer's instructions. Plasma cortisol was measured using DPC Immulite 2000 using a solid phase 2 site chemiluminescent immunometric assay. Results: Mean plasma cortisol levels were 400 nmol/L ± 42.8 SEM, and mean salivary cortisol levels were 127.5 nmol/L ± 66.5 SEM. Plasma cortisol was positively correlated with salivary cortisol (r=0.41, p<0.001). Conclusion: The study showed a reasonable correlation between salivary and plasma cortisol concentrations obtained early in the morning in the extremely preterm infants.