Daily cortisol awakening response and menstrual symptoms in young females.

Menstrual symptoms include some rhythmical changes and stress perception but women differ in duration and severity of these symptoms. It is not known whether these differences are associated with cortisol awakening response (CAR). The aim of the current study was to follow young women daily for the CAR and menstrual parameters throughout the whole menstrual cycle. Healthy and regularly cycling young women (n = 16, 17 to 31-year-old) participated in the current study. The daily records of severity of problems (DRSP) was filled in daily by the participants. CAR was also assessed daily form the salivary samples collected at a 0-, 15-, 30- and 60-min post-awakening. In terms of daily awakening cortisol profiles, women had either 2-20 (n = 3), or 20-200 (n = 8) or 200-2000 (n = 5) ng/ml cortisol according to the median levels throughout their cycle. CAR was weakly and negatively correlated with DRSP scores but strongly and positively with oestradiol (R2  = 0.300; p = 0.000) and progesterone (R2  = 0.490; p = 0.000) concentrations. Individuals with higher oestradiol and progesterone concentrations did not have high DRSP scores. In conclusion, CAR had a very high between-subject difference but had a low within-subject difference throughout the days of menstrual cycle, suggesting that CAR is a relatively stable personal trait. Moreover, the interplay between cortisol, progesterone and oestradiol appears to be important for the severity of menstrual symptoms.

The hypothalamic-pituitary-adrenal axis activity in archers: cortisol release, stress, anxiety and success.

BACKGROUND: Archery is a unique sport requiring simultaneous physical and psychological focusing for precisely hitting the small, distant target. Thus, in this sport, the effects of stress and anxiety on success might be more pronounced as a slight deviation in aiming may translate into a large error in meeting the target. Therefore, the current study aimed to assess the interrelationship between anxiety, cortisol awakening response (CAR), cortisol levels during the shooting period, and success in professional male archers during a national tournament. METHODS: Archers (16-20 years old, male, N.=20) shooting with recurve bow participated in the current study during the indoor archery championship. For the assessment of CAR, salivary samples were collected at 0- (wake up), 30-, 45- and 60-min postawakening on three consecutive days, namely qualification, individual elimination, and team shooting days. On the first two days of the shootings (i.e., qualification and elimination), shooting salivary samples were collected at 30 and 15 minutes before the shootings, at half-time and just after the shootings. State and trait anxiety inventory were filled in on the day of registration (the day before qualification shootings). RESULTS: CAR, measured as the area under the curve (AUC), was higher on the elimination day (P=0.038) compared to the qualification day. Shooting cortisol levels were also higher on the elimination day compared to the qualification day (P=0.004). Archers having high rankings on qualification day also had higher success rates on elimination day (r=0.963, P<0.001). CONCLUSIONS: Moderately increased state anxiety, higher CAR, and elevated shooting cortisol levels appear to be the integral components of the elimination stage during a real competition in archery. Thus, assessment of secretion dynamics of cortisol appears to be a valuable objective tool for understanding the neuroendocrine control during the competition days.

Analysis of long-term antibody response in COVID-19 patients by symptoms grade, gender, age, BMI, and medication.

The first aim of the study was to analyze the change in antibody titer at 15-day intervals until 60 days postsymptom onset (PSO). The second aim was to analyze the relationship between antibody titer and symptom grade, gender, age, body mass index (BMI), medications, vitamin supplements, and herbal therapies. Blood samples were collected from 43 patients (5 mild, 21 moderate, 17 severe diseases), 18 women (41.9%), and 25 men (58.1%), on 15, 30, 45, and 60 days PSO after COVID-19 infection. The serum antibody titers were determined by measuring the COVID-19 immunoglobulin G (IgG) antibodies by enzyme-linked immunoassay (ELISA). Associations between the duration of symptoms, demographic and clinical parameters, medications and vitamins used, and herbal therapies were evaluated by interviewing the participants. Within the first 15 days of illness, 81.4% of the patients were positive. From Day 45 PSO, seropositivity was 89.5%. The anti-SARS-CoV-2 antibody titers were statistically higher in men than women at all times (p < 0.01). Antibody titer was higher in older participants compared to younger participants (p < 0.02). Plaquenil or favipiravir use did not affect antibody response (p > 0.05). Men had a higher fever (p = 0.006), shortness of breath (p = 0.004), and chest pain (p = 0.03) than women. We found powerful antibody response by 60 days PSO, as well as higher antibody response and severity of symptoms in the men gender. Data also showed that SARS-CoV-2 antibodies are higher in individuals with older age, whereas BMI, concomitant chronic disease, and medications had no effect on antibody titers.

Effects of late-night eating of easily-or slowly-digestible meals on sleep, hypothalamo-pituitary-adrenal axis, and autonomic nervous system in healthy young males.

Aim of the current study was to assess the effects of the digestibility of late-night high calorie meal on sleep and the activities of the hypothalamo-pituitary-adrenal (HPA) and autonomous nervous system (ANS) in healthy young males. For that purpose, effects of an easily digestible meal (starch + sugar-rich meal, SSR, i.e., dessert) or a slowly digestible meal (protein + fat-rich, PFR, i.e. kebab) were investigated in a crossover design in 16 participants (20-26 year old). They did not eat anything after 07:00 PM on Day 0; had an SSR meal on Day 1 and a PFR meal on Day 2 at 10:00 PM. HPA and ANS activities were measured by cortisol awakening response (CAR) and heart rate variability (HRV), respectively. The participants provided salivary samples for CAR; had a 5-min continuous electrocardiogram recording for HRV; and filled in sleep questionnaires. Late-night eating of SSR and PFR diets increased the area under the curve of CAR (p < 0.05) but did not affect HRV parameters (p > 0.05). PFR meal significantly disturbed sleep (p < 0.05). The data suggests that increased activity of HPA, but not ANS, might be involved in pathophysiology of late-night eating and that this might be due to disturbed sleep if slowly-digestible meal is consumed.

Cortisol Awakening Response and Heart Rate Variability in the Menstrual Cycle of Sportswomen.

Purpose: This study aimed to examine the activities of heart rate variability (HRV) and cortisol secretion as markers of the autonomous nervous system (ANS) and the hypothalamic-pituitary-adrenal axis (HPA) during the menstrual cycle in female athletes. Method: Saliva samples of the participants (n = 28) were collected successively at 0, 15, 30, and 60 min after awakening to assess cortisol awakening response (CAR) during each of four phases of the menstrual cycle (the menstruation, ovulation, luteal, and premenstrual phases). Diurnal saliva samples were collected at noon, in the evening (17:00) and 22:00 for cortisol analysis. HRV was measured in the menstrual and premenstrual phases at rest (12:00-14:00). Results: There was no difference between CAR parameters in the menstrual phases. CAR parameters and diurnal cortisol levels were significantly correlated with each other throughout the menstrual cycle. Mean cortisol was a strong predictor of the area under the curve (AUC: S = 0.042; R-Sq = 98.4%). Time-dependent parameters of HRV (standard deviation of the N-N interval [SDNN; P = .049] and the root mean square of the successive R-R differences [rMSSD; P = .038]) were significantly higher in the menstrual phase. There were no correlations between CAR and HRV parameters. Conclusions: We suggest that the hypothalamic-pituitary-adrenal axis activity and the autonomic nervous system work in a separate manner that is not affected by the menstrual phases in sportswomen. The present study contributed to a better understanding of the relationship between the dynamics of cortisol release and the autonomic nervous system in different phases of the menstrual cycle in female athletes.

Stress in couples undergoing assisted reproductive technology.

PURPOSE: To determine the associations between pre-treatment self-reported stress level, salivary cortisol levels, and clinical pregnancy outcome in couples undergoing assisted reproductive technology treatment (ART). STUDY DESIGN: Seventy-five couples (150 patients) undergoing ART treatment were enrolled in this study. Psychological variables were assessed using the Perceived Stress Scale, Beck Anxiety Inventory, State-Trait Anxiety Inventory, and Beck Depression Inventory. Salivary cortisol levels were obtained from each couple prior to commencing gonadotropin treatment at several times (upon waking and at 15, 30, and 60 min after waking). RESULTS: There was no statistically significant association between the self-reported stress levels and the ART treatment outcome in couples. Women with a successful outcome after ART treatment had higher median salivary cortisol levels than women who had an unsuccessful result [24.7 (19.9-63.1) vs. 20.7 (10.4-30.4), respectively]. There was no statistically significant difference in the median salivary cortisol levels of men in relation to ART treatment outcome. Salivary cortisol levels of the couples were not correlated. CONCLUSIONS: Women who had higher median salivary cortisol levels in the pre-treatment period had a higher clinical pregnancy rate. This result suggests that moderately increased activity of the hypothalamic-pituitary-adrenal axis during ART treatment might be associated with successful conception.

Does Psychodrama Affect Perceived Stress, Anxiety-Depression Scores and Saliva Cortisol in Patients with Depression?

OBJECTIVE: Little is known about the effects of psychodrama group therapy on the level of stress, anxiety and depression of the group members, and the biological markers of stress. The aim of this study test cortisol level as a biological reflection of some mental characteristics gained by the psychodrama method in coping with stress. METHODS: Depressive patients aged between 18 and 65 years, who diagnosed with depression according to DSM-5, without psychiatric comorbidity, who do not use drugs and were available for psychodrama were enrolled into the study. These patients were evaluated using the Perceived Stress Scale, State-Trait Anxiety Inventory and Beck Depression Inventory. In order to observe the circadian rhythm of cortisol, samples were collected from the participants before lunch, before dinner and before going to sleep on the day before psychodrama and on the 0, 15th, and 30th minutes after awakening in the morning of the psychodrama day; as well as just before and after the psychodrama session. Saliva cortisol level just before the initial session of psychodrama group therapy was compared with the saliva cortisol level just after the last psychodrama session at the end of 16 weeks. RESULTS: Statistically significant difference was determined between the scores of perceived stress scale, STAI-1, STAI-2, beck depression inventory and salivary cortisol level before and after psychodrama session. There was significant decrease in both the scale scores and salivary cortisol after vs. before psychodrama. CONCLUSION: This results is important as it shows the biological aspect of clinical improvement. Further studies would provide us with better understanding of the effects of psychodrama group psychotherapy on depressive mood and biological projections by means of short-term and long-term follow-up studies.

Maternal viral mimetic administration at the beginning of fetal hypothalamic nuclei development accelerates puberty in female rat offspring.

This study aimed to investigate the effects of maternal viral infection during a critical time window of fetal hypothalamic development on timing of puberty in the female offspring. For that purpose, a viral mimetic (i.e., synthetic double-strand RNA, namely, polyinosinic-polycytidylic acid, poly (I:C)) or saline was injected (i.p.) to the pregnant rats during the beginning (day 12 of pregnancy, n = 5 for each group) or at the end of this time window (day 14 of pregnancy, n = 5 for each group). Four study groups were formed from the female pups (n = 9-10 pups/group). Following weaning of pups, vaginal opening and vaginal smearing was studied daily until 2 sequential estrous cycles were observed. During the second diestrus phase, blood samples were taken for progesterone, leptin, corticosterone, follicle-stimulating hormone, and luteinizing hormone. Maternal poly (I:C) injection on day 12 of pregnancy increased body mass and reduced the time to puberty in the female offspring. Neither poly (I:C) nor timing of injection affected other parameters studied (p > 0.05). It has been shown for the first time that maternal viral infection during the beginning of fetal hypothalamic development might hasten puberty by increasing body mass in rat offspring.

Late-night exercise affects the autonomic nervous system activity but not the hypothalamo-pituitary-adrenal axis in the next morning.

BACKGROUND: Exercise activates hypothalamic-pituitary-adrenal axis (HPA) and autonomic nervous system (ANS) and generally causes beneficial changes in homeostatic balance. However, the health benefits of late-night exercise programs on the activity of HPA and ANS is not known. The aim of this study was to assess effects of late-night exercise on sleep quality and on the activities of the HPA axis (as cortisol awakening response, CAR) and the ANS (as heart rate variability, HRV) measurements in the following morning. METHODS: Medical students (N.=20 males, 20-24 years old) filled Karolinska Sleep Diary on the day before exercise program. In the following morning, they provided salivary samples for the assessment of CAR (samples at 0, 15, 30 and 60 min postawakening) and had a 5-min electrocardiogram recording for the determination of HRV. In the next night, an exercise program consisting of a 90-min football match was implemented at 09:30 p.m. and all procedures were repeated. Cortisol concentrations were measured in the salivary samples and time- and frequency-domain parameters of HRV were calculated. RESULTS: Late-night exercise did not affect (P>0.05) sleep parameters (sleep duration, disturbed sleep, awakening problems) and CAR parameters (0, 15, 30, 60 min cortisol concentrations, mean concentration, area under the curve) but influenced HRV parameters (P<0.05). CONCLUSIONS: The results suggest that late-night exercise is associated with changed HRV activity rather than changes in CAR and, therefore, it might be suggested that late-night exercise affects ANS activity rather than HPA activity in the next morning.

ACA, an inhibitor phospholipases A2 and transient receptor potential melastatin-2 channels, attenuates okadaic acid induced neurodegeneration in rats.

AIM: In recent studies, it has been shown that the Transient Receptor Potential Melastatin-2 Channels (TRPM2) and Phospholipases A2 (PLA2) inhibitors may have a protective effect on neurons. This study was aimed to investigate the protective effect of TRPM2 and PLA2 inhibitor N-(p-amylcinnamoyl) Anthranilic Acid (ACA) in a neurodegenerative model induced by Okadaic Acid (OKA). MAIN METHODS: OKA (200ng/10µl) was administered bilateral intracerebroventricularly as a single injection. KEY FINDINGS: OKA-treated rats showed significant impairments of spatial memory in Morris Water Maze Test. OKA-induced memory-impaired rats showed increased numbers of degenerated neurons and Caspase-3, tau phosphorylated ser396, ß-amyloid positive cells in the hippocampus and cerebral cortex. Furthermore, OKA-treated rats exhibited significantly increased MDA, TNF-α levels, and decreased SOD, GSH-PX enzyme activates and GSH levels of the tissues. ACA administration ameliorated OKA-induced memory impairment in rats. The ACA treatment also increased SOD and GSH-PX enzyme activation and GSH levels, and conversely decreased the levels of MDA, TNF-α. It was found that the numbers of the degenerated neurons and Caspase-3 positive cells of cortex and hippocampus regions were significantly reduced. SIGNIFICANCE: ACA administration attenuates the oxidative stress and neuroinflammation of OKA-induced neurodegeneration; and ameliorates the cognitive decline and neurodegeneration.

Cortisol awakening response is blunted and pain perception is increased during menses in cyclic women.

BACKGROUND AND AIMS: The incidence of menstrual symptoms is reported to be as high as 90% in cyclic women. These symptoms, including anxiety and pain, might be associated with cortisol, as its receptors are widely distributed in the brain areas associated with behavior. Therefore, the current study aimed to assess the cortisol awakening response (CAR) throughout the menstrual cycle and correlate it with pain perception and trait anxiety. MATERIALS AND METHODS: CAR was assessed by measuring salivary cortisol at 0, 15, 30, and 60min following awakening in the same women (n=59, age 22.2±0.37years) at various stages of the menstrual cycle (menses, midcycle, luteal and premenstrual phases). Progesterone and estradiol concentrations were also determined in saliva samples to assess cyclic changes. Self-reported pain, trait anxiety, and menstrual symptoms were assessed by visual analog scale (VAS), state-trait anxiety inventory (STAI-T), and the Daily Record of Severity of Problems (DRSP), respectively. RESULTS: Estradiol was significantly elevated during the midcycle period and remained high during the early luteal phase (p<0.05). Progesterone was increased during the luteal phase (p<0.05). Post-awakening cortisol values increased during midcycle, luteal phase, and premenstrual phase (p<0.05, classical CAR), but not during the menses (p>0.05, blunted or flat CAR). Positive and significant correlations were found between cortisol and estradiol (R2=0.322; p=0.000), cortisol and progesterone (R2=0.156; p=0.000), and estradiol and progesterone (R2=0.349; p=0.001). Premenstrual symptom scores were higher in the menses and premenstrual phases than in the midcycle and luteal phases (p<0.001). Pain perception was the highest during the menses followed by the premenstrual phase (p<0.01). CONCLUSIONS: CAR was blunted during the menses, suggesting that cortisol might play a phase-specific role in the regulation of the cycle. Additionally, premenstrual symptoms, including pain, were more severe when ovarian steroid levels reduced (i.e., menses and the premenstrual phase).

Development and validation of an enzyme-linked immunosorbent assay for detection of cortisol in human saliva.

Non-invasive measurement of cortisol in saliva is of prime importance as it represents a bioavailable neuroendocrine marker for stress. Therefore, in this study, we developed an enzyme immune assay that was suitable for salivary cortisol measurements. For that purpose, rabbit polyclonal antibody was raised against cortisol-3-CMO:BSA conjugate. The test was based on competition of liquid phase cortisol with conjugated cortisol on the solid phase. Primary antibody was used to bind available sites on the conjugate, which was proportional to numbers of cortisol in liquid phase. Biotinylated secondary anti-rabbit antibody was used to detect primary antibodies by addition of streptavidin peroxidase and substrate, respectively. Color formation was stopped and yellow color was read by a plate-reader spectrophotometer. Additionally, validated test was used to met all validation criteria including. Test developed was used to establish cortisol awakening response (CAR) in saliva samples collected in the morning after awakening (0, 15, 30, and 60th min) from women (n = 4) and men (n = 4) at 8 or 4 different days, respectively. Diurnal cortisol levels were assessed (n = 8) at after awaking 60 min at morning, 12:00, 19:00, and 22:00 hr. In conclusion, an enzyme immunoassay test was successfully produced, validated and used for cortisol measurement in saliva samples.

Endotoxin exposure and puberty in female rats: the role of nitric oxide and caspase-1 inhibition in neonates.

Bacterial toxins are widespread in the environment as well as in the digestive system of humans and animals. Toxin from Gram-negative bacteria (endotoxin or lipopolysaccharide; LPS) has a life-long programming effect on reproduction in rats, but the mediators have not been well-documented, so we investigated the effects of LPS on the timing of puberty in female rats. Because the levels of nitric oxide (NO) and interleukin 1ß (IL-1ß) increase following injection of LPS, we injected neonates (post-natal day (pnd) 7) with LPS, with or without NO or IL-1ß inhibitors. Half of the prepubescent (pnd 30) animals received an additional LPS injection. Vaginal opening, number of ovarian follicles, and serum anti-LPS antibodies were determined. A single LPS injection was sufficient to reduce the primordial follicle pool, but puberty was delayed when rats received 2 LPS injections (at pnd 7 and 30). NO or IL-1ß inhibitors improved both of these parameters, suggesting that the early detrimental effects of LPS on puberty and primordial follicle pool are mediated by NO and IL-1ß.

Kisspeptin antagonist prevents RF9-induced reproductive changes in female rats.

The aim of this study was to determine the modulatory effects of peptide 234 (p234) (an antagonist of GPR54 receptors) on kisspeptin and RF9 (an RFamide-related peptide antagonist)-induced changes in reproductive functions and energy balance in female rats. Female Sprague-Dawley rats were weaned on postnatal day (pnd) 21. The animals were intracerebroventricularly cannulated under general anesthesia on pnd 23. Groups of female rats were injected with kisspeptin, RF9, p234, kisspeptin plus p234, or RF9 plus p234, daily. The experiments were ended on the day of first diestrus following pnd 60. Kisspeptin or RF9 alone advanced vaginal opening (VO), which was delayed by administration of kisspeptin antagonist alone. In the rats given kisspeptin plus p234 or RF9 plus p234, VO was not different from control rats. Kisspeptin and RF9 elicited significant elevations in circulating LH levels. Coadministrations of kisspeptin or RF9 with p234 decreased LH levels significantly. The use of p234 alone did not cause any significant change in LH secretion. Kisspeptin decreased both food intake and body weight while RF9 decreased only food intake without affecting body weight. The effects of kisspeptin on energy balance were also reversed by central administration of p234. In conclusion, kisspeptin antagonist, p234, modulates the effects of kisspeptin on reproductive functions and energy balance, whereas RF9 seems to exert only its effects on reproductive functions by means of GPR54 signaling in female rats.