Total Cortisol Concentrations Research Articles

Detection of endogenous cortisol in equine tears and blood at rest and after simulated stress

To determine whether cortisol is present in equine tears at rest and during simulated stress and compare tear cortisol to serum free and total cortisol. Fourteen healthy adult horses were included. Paired tear total cortisol and serum total and free cortisol concentrations were measured with ELISA, chemiluminescent immunoassay, and ultrafiltration methodology, respectively, in 10 horses at rest once daily for five consecutive days. In an additional four horses, paired tear and serum samples were collected for cortisol measurement before and after adrenocorticotropic hormone (ACTH) stimulation (cosyntropin, 1 μg/kg IV). Cortisol was detectable in equine tears at rest. Following ACTH stimulation, tear cortisol increased significantly from baseline at 60-120 min (P ≤ 0.001). Serum total and free cortisol also increased significantly at 30-180 min after ACTH stimulation (P ≤ 0.001). Both serum and tear cortisol returned to baseline concentrations by 360 min. Changes in tear cortisol were similarly associated with changes in serum total and free cortisol, although high tear cortisol concentrations suggest a portion of tear cortisol may be protein-bound. Cortisol is present in equine tears and increases in concert with serum cortisol following ACTH stimulation. Further study is needed to determine whether endogenous cortisol in tears contributes to ocular pathology.

Gender determines ACTH recovery from hypercortisolemia in healthy older humans

ObjectiveAvailable clinical data raise the possibility that stress-adaptive mechanisms differ by gender. However, this notion has not been rigorously tested in relation to cortisol-mediated negative feedback. Materials/MethodsDegree of ACTH inhibition during and recovery from an experimental cortisol clamp was tested in 20 healthy older subjects (age 60±2.2y). Volunteers received oral placebo or ketoconazole (KTCZ) to inhibit adrenal steroidogenesis along with i.v. infusions of saline or a low vs high physiological dose of cortisol in a prospectively randomized double-blind, placebo-controlled design. ACTH and cortisol concentrations were measured every 10min during the feedback-clamp phase and thereafter (recovery or escape phase). Corticosteroid-binding globulin (CBG) was measured, and free cortisol concentrations were calculated. ResultsGender did not determine mean ACTH concentrations during the saline or cortisol feedback-clamp phases per se. However, women had markedly impaired ACTH recovery after stopping both low- and high-dose cortisol infusions compared with men (P=0.005, KTCZ/low-dose cortisol arm; and P=0.006, KTCZ/high-dose cortisol arm). Decreased ACTH recovery in women was accompanied by lower total and free cortisol concentrations, pointing to heightened feedback inhibition of hypothalamo-pituitary drive of ACTH secretion as the main mechanism. ConclusionsIn summary, gender or a factor related to gender, such as sex steroids or body composition, determines recovery of ACTH secretion from cortisol-enforced negative feedback. Attenuated ACTH recovery in post-menopausal women may have relevance to sex differences in stress-related adaptations.

Repeated mixing and isolation: Measuring chronic, intermittent stress in Holstein calves

Objectives of this study were to determine the physiological effects of psychological stress applied to dairy calves and to test if molasses consumption could be used to validate that a stressed condition was achieved. Twenty male calves (3wk old) received jugular catheters and were randomly assigned to control (CTR; n=4 pens of 1 calf per pen) or social stress treatments (STR; n=4 pens of 4 calves per pen). The STR treatment included 5 cycles of 24-h isolation followed by regrouping with unfamiliar animals for 48h (over 15d). An ACTH challenge (0.1 IU/kg of body weight) was used to determine adrenal fatigue. Peak and total cortisol concentrations were greater for STR calves until the ACTH challenge. After the ACTH challenge, CTR calf cortisol increased and STR calf cortisol continued to decrease, suggesting adrenal fatigue. The number of calves that became positive for fecal shedding of Salmonella after the acute stress of being moved and the number of calves that were positive after the move decreased with each move. Fifty-six percent of STR calves changed from negative to positive for shedding after the first move compared with 18.75% of STR calves remaining negative after the third move. Difference in fecal shedding of Enterobacteriaceae from samples taken before and after moving calves on d 6 was less than that on d 2, 3, and 5. Leukocyte counts were not different, but trends for day effects were detected for neutrophil and monocyte percentages. Molasses consumption was greater for STR calves on d 2 and 11, as was total consumption. Latency to lie after eating also increased as the study progressed; STR calves required more time to lie after eating on d 12 than on d 3, and latency to lie was greater for STR than CTR on d 4, 8, 12, and 14. The STR calves also stood more than the CTR calves in the 4-h afternoon period on d 4, 5, 7, and 14. However, during the 4-h morning observations on d 14 (ACTH challenge), CTR calves stood more than STR calves. This model induced chronic stress, as characterized by adrenal fatigue, which was confirmed by molasses consumption and behavior changes. Therefore, molasses consumption could be used to confirm social stress in experimental models.

Comparison of Total Cortisol, Free Cortisol, and Surrogate Markers of Free Cortisol in Diagnosis of Adrenal Insufficiency in Patients With Stable Cirrhosis

Measurements of serum levels of total cortisol can overestimate the prevalence of adrenal dysfunction in patients with cirrhosis because they have low concentrations of corticosteroid-binding globulin and albumin. We used measurements of serum total cortisol and serum free cortisol after the low-dose short Synacthen test (LDSST) to assess adrenal dysfunction. We studied 79 patients with stable cirrhosis; adrenal dysfunction was defined by peak concentrations of total cortisol ≤494 mmol/L and/or peak concentrations of free cortisol ≤33 nmol/L after the LDSST. We determined free cortisol index (FCI) scores and calculated free cortisol levels by using Coolens' equation. The Cox regression model was used to assess the relationship between adrenal dysfunction and outcomes (death or liver transplant). On the basis of measurement of total cortisol, 34% of patients had adrenal dysfunction, and on the basis of measurement of free cortisol, 29% had adrenal dysfunction. There was agreement between total cortisol and free cortisol levels in 22% of patients; in 13%, adrenal dysfunction was diagnosed only on the basis of total cortisol and in 6% only on the basis of free cortisol (κ coefficient, 0.56; P < .01). Low concentrations of corticosteroid-binding globulin (21 vs 54 μg/mL, P < .01) led to an overestimation of adrenal dysfunction that was based on measurement of total cortisol. Measurements of calculated free cortisol constantly overestimated free cortisol concentrations, with variations as large as 87% for baseline values and up to 84% after stimulation. Adrenal insufficiency, defined by FCI scores <12, was detected in 30% of patients; among them, 23% also had subnormal peak levels of free cortisol (κ coefficient, 0.70; P < .001). Adrenal dysfunction was not significantly associated with patient outcomes, on the basis of Cox model analysis. Adrenal insufficiency, defined by LDSST, is frequent in patients with stable cirrhosis, on the basis of measurements of total and free cortisol. FCI scores are better than measurement of total cortisol in assessing adrenal function in patients with cirrhosis. We did not associate adrenal dysfunction with outcome, but further studies are needed.

Relation of inflammation and liver function with the plasma cortisol response to adrenocorticotropin in early lactating dairy cows

In this study we examined the relationship between cortisol and inflammatory status in early lactating dairy cows after a stimulation test of the adrenal cortex. Twenty-four cows were grouped into quartiles (6 cows per each quartile) in accordance with the liver activity index (based on plasma concentration of negative acute phase proteins in early lactation); the quartiles were lower (LO; cows with the lowest liver functionality), intermediate lower, intermediate upper, and upper (UP; cows with the highest liver functionality). Each cow was injected i.v. with 20µg of a synthetic analog of ACTH at 35d in milk (DIM). Blood samples were taken to assess inflammatory status, and at 0, 30, and 60min after ACTH challenge to measure total cortisol. The free cortisol fraction was analyzed in the LO and UP quartiles and the bound cortisol fraction was estimated as the difference between total and free cortisol. The LO, in comparison with the other quartiles, suffered a more severe inflammatory status, with the highest values of haptoglobin, reactive oxygen metabolites, and total nitric oxide metabolites and the lowest concentration of direct or indirect markers of negative acute phase proteins. The cows in the LO quartile had the highest values of plasma nonesterified fatty acids and β-hydroxybutyrate at 7 DIM, suggesting a more severe body lipid mobilization. The LO quartile cows showed the highest frequency of health problems and the lowest milk yield in the first 35 DIM. Thirty minutes after the ACTH treatment, the concentration of total cortisol was lower in LO in comparison to other groups. Similarly, the bound cortisol fraction was lower in LO versus UP. The adrenal response appeared inversely related with health status after calving (e.g., lower in LO cows, experiencing the most severe inflammatory status). The lower increase in cortisol after the ACTH challenge in cows with greater inflammation (LO quartile) seems a consequence of the lower availability of cortisol-binding globulin synthetized by the liver, but other mechanisms can be involved (e.g., rate of cortisol production, secretion, and metabolic clearance). Our data provide evidence that inflammation and metabolic changes reduce the concentration of circulating plasma cortisol during an acute stress. Hence, the acute phase response in dairy cows should be taken into account to interpret the results obtained from stimulation tests of the adrenal cortex.

Development and validation of a sensitive enzyme immunoassay (EIA) for blood plasma cortisol in female cattle, buffaloes, and goats

A highly sensitive enzyme immunoassay (EIA) that used the second antibody coating technique and the cortisol-horseradish peroxidase conjugate as a label for determination of free and total cortisol in blood plasma of dairy animals (cows, buffaloes, and goats) was developed. For biological validation of the EIA, blood samples were collected from the animals at 48 and 24 h before and 0, 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, and 132 h after dexamethasone administration. The EIA was performed directly with 20 μL of fresh plasma (for free cortisol) and also with 20 μL of heat-treated plasma (for total cortisol) after 1:5 dilutions with PBS. Cortisol standards ranging from 0.39 to 200 pg/well/20 μL were used, and the sensitivity of the EIA procedure was found to be 0.39 pg/well/20 μL, which corresponded to 0.02 ng/mL. In comparison with RIA the EIA was at least 4 times more sensitive and required 5 times less cortisol antiserum. In female cattle, buffaloes, and goats, the total, free, and bound plasma cortisol before dexamethasone administration was significantly (P < 0.05) higher than the total, free, and bound cortisol after dexamethasone administration. It can be concluded from these studies that the direct, sensitive EIA validated for estimating the free and total cortisol concentrations was sufficiently reliable and quick for studying the dynamics of cortisol distribution in blood plasma of dairy animals.

Pre-Operative and Early Post-Operative Factors Associated with Surgical Site Infection after Laparoscopic Sleeve Gastrectomy

Surgical procedures on obese patients are expected to have a high incidence of surgical site infection (SSI). The identification of pre-operative or early post-operative risk factors for SSI may help the surgeon to identify subjects in risk and adequately optimize their status. We conducted a study of the association of comorbidities and pre- and post-operative analytical variables with SSI following laparoscopic sleeve gastrectomy for the treatment of morbid obesity. We performed a prospective study of all morbidly obese patients undergoing laparoscopic sleeve gastrectomy as a bariatric procedure between 2007 and 2011. An association of clinical and analytical variables with SSI was investigated. The study included 40 patients with a mean pre-operative body mass index (BMI) of 51.2±7.9 kg/m(2). Surgical site infections appeared in three patients (7.5%), of whom two had an intra-abdominal abscess located in the left hypochondrium and the third had a superficial incisional SSI. Pre-operatively, a BMI >45 kg/m(2) (OR 8.7; p=0.008), restrictive disorders identified by pulmonary function tests (OR 10.0; p=0.012), a serum total protein concentration <5.3 g/dL (OR 13; p=0.003), a plasma cortisol >30 mcg/dL (OR 13.0; p=0.003), and a mean corpuscular volume (MCV) <82 fL (OR 1.6; p=0.04) were associated with post-operative SSI. Post-operatively, a serum glucose >128 mg/dL (OR 4.7; p=0.012) and hemoglobin <11g/dL (OR 7.5; p=0.002) were associated with SSI. The study supports the role of restrictive lung disorders and the values specified above for preoperative BMI, serum total protein and cortisol concentrations, and MCV, and of post-operative anemia and hyperglycemia as risk factors for SSI. In these situations, the surgeon must be aware of and seek to control these risk factors.

Defining adrenal status with salivary cortisol by gold-standard insulin hypoglycemia

BackgroundInsulin-induced hypoglycemia (IHT) is considered the gold standard test for evaluating the HPA axis. Serum free cortisol or its surrogate, salivary cortisol as opposed to total cortisol concentrations, offers a better reflection of the activation of HPA axis. Our study aimed to derive reference ranges for the normal salivary cortisol levels in healthy patients and patients with adrenal insufficiency. Design and methodsSerum cortisol concentrations, using the gold standard of IHT, and salivary cortisol were obtained. 36 patients referred to our outpatient endocrine testing unit for evaluation of adrenal function were included in the study. Most subjects had a history of suspected hypothalamic/pituitary disease causing adrenal insufficiency. ResultsWe found a strong linear correlation between the serum and salivary cortisol concentrations in simultaneously collected samples (r=0.81, 95% CI 0.74–0.86, p<0.0001). The corresponding salivary cortisol equivalent to a serum cortisol of 500nmol/L, using a linear-regression equation, was 16.7nmol/L (95% CI 13.3–20.1nmol/L, p=0.0001). A salivary cortisol of 13.3nmol/L has a specificity of 89.3% to detect abnormal HPA function. Using the upper 95% CI result of salivary cortisol 20.1 yields a sensitivity of 87.5%. ConclusionWith the present assay, adrenal insufficiency may be diagnosed with reasonable confidence if a random salivary cortisol is lower than 13.3nmol/L and excluded if a random salivary cortisol is higher than 20.1nmol/L. Future studies should correlate these thresholds with clinical outcomes.

Monitoring the circadian rhythm of serum and salivary cortisol concentrations in the horse

Daily fluctuations of cortisol concentration in the blood or saliva have been repeatedly reported. However, several contradictions in the existing literature appear on this subject. The present study was performed to definitively establish options for testing adrenocortical function. To the best of our knowledge, this is the first study to evaluate parallel circadian rhythms in salivary and serum cortisol concentrations during a 24-h period. Twenty horses were examined under the same conditions. Blood and saliva samples were taken every 2 h for 24 h to determine the daily changes in cortisol concentrations of saliva and serum at rest and to determine the relationship between salivary and serum cortisol levels. Cosinor analysis of group mean data confirmed a significant circadian component for both serum and salivary cortisol concentrations (P < 0.001 in both cases). The serum cortisol circadian rhythm had an acrophase at 10:50 AM (95% CI, 10:00 AM–11:40 AM), a MESOR of 22.67 ng/mL, and an amplitude of 11.93 ng/mL. The salivary cortisol circadian rhythm had an acrophase at 10:00 AM (95% CI, 9:00 AM–11:00 AM), a MESOR of 0.52 ng/mL, and an amplitude of 0.12 ng/mL. We found a significant but weak association between salivary and serum cortisol concentrations; the Pearson correlation coefficient was 0.32 (P < 0.001). The use of salivary cortisol level as an indicator of hypothalamic-pituitary-adrenal axis activity may be warranted. However, the salivary cortisol levels are more likely to be correlated with free plasma cortisol than with the total plasma cortisol concentration.

Reduced Glucocorticoid Receptor Protein Expression in Children with Critical Illness

Background/Aims: The diagnostic criteria for critical illness-related corticoid insufficiency (CIRCI) are not well established, particularly for children. In addition to alterations in adrenal function, cellular resistance to glucocorticoid action could contribute to CIRCI due to alterations in the functioning of the intracellular receptor protein for corticosteroids, the glucocorticoid receptor (GR). Methods: We have therefore undertaken a pilot, prospective study to assess whether cellular GR activity can be measured in peripheral blood mononuclear cells (PBMCs) from critically ill children. Results: Total and cytoplasmic, but not nuclear GR levels were significantly lower in PBMCs from critically ill children (i.e. sepsis/septic shock and traumatic brain injury) compared to healthy controls. While total cortisol concentrations did not differ between test groups, salivary and serum-free cortisol concentrations were significantly greater in both groups of children with critical illness. Cortisol-binding globulin levels were significantly lower in patients with sepsis/septic shock. Conclusions: The lower total and cytoplasmic receptor levels in critically ill children suggest that the GR-mediated response to exogenous glucocorticoid therapy may be limited. However, the nuclear transport of GR in critically ill patients suggests that residual receptors in these patients retain functionality and may be accessible to therapeutic treatments that maximize their activity.

Influence of upper-body exercise order on hormonal responses in trained men

The aim of this study was to examine acute hormonal responses after different sequences of an upper-body resistance-exercise session. Twenty men completed 2 sessions (3 sets; 70% 1-repetition maximum; 2 min passive rest between sets) of the same exercises in opposite sequences (larger to smaller vs. smaller to larger muscle-group exercises). Total testosterone (TT), free testosterone (FT), testosterone/cortisol (T/C) ratio, sex-hormone-binding globulin (SHBG), growth hormone (GH), and cortisol (C) concentrations were measured before and immediately after each sequence. The results indicate that the GH concentration increased after both sessions, but the increase was significantly greater (p < 0.05) after the sequence in which larger muscle-group exercises were performed prior to the smaller muscle-group exercises. No differences were observed between sessions for TT, FT, SHBG, C, or the T/C ratio at baseline or immediately after resistance exercise. These results indicate that performing larger muscle-group exercises first in an upper-body resistance-exercise session leads to a significantly greater GH response. This may have been due to the significantly greater exercise volume accomplished. In summary, the findings of this investigation support the common prescriptive recommendation to perform larger-muscle group exercises first during a resistance-exercise session.

High and low protein∶ carbohydrate dietary ratios during gestation alter maternal-fetal cortisol regulation in pigs.

Imbalanced maternal nutrition during gestation can cause alterations of the hypothalamic-pituitary-adrenal (HPA) system in offspring. The present study investigated the effects of maternal low- and high-protein diets during gestation in pigs on the maternal-fetal HPA regulation and expression of the glucocorticoid receptor (GR), mineralocorticoid receptor (MR), 11β-hydroxysteroid dehydrogenase 1 and 2 (11β-HSD1 and 11β-HSD2) and c-fos mRNAs in the placenta and fetal brain. Twenty-seven German Landrace sows were fed diets with high (HP, 30%), low (LP, 6.5%) or adequate (AP, 12.1%) protein levels made isoenergetic by varying the carbohydrate levels. On gestational day 94, fetuses were recovered under general anesthesia for the collection of blood, brain and placenta samples. The LP diet in sows increased salivary cortisol levels during gestation compared to the HP and AP sows and caused an increase of placental GR and c-fos mRNA expression. However, the diurnal rhythm of plasma cortisol was disturbed in both LP and HP sows. Total plasma cortisol concentrations in the umbilical cord vessels were elevated in fetuses from HP sows, whereas corticosteroid-binding globulin levels were decreased in LP fetuses. In the hypothalamus, LP fetuses displayed an enhanced mRNA expression of 11β-HSD1 and a reduced expression of c-fos. Additionally, the 11β-HSD2 mRNA expression was decreased in both LP and HP fetuses. The present results suggest that both low and high protein∶carbohydrate dietary ratios during gestation may alter the expression of genes encoding key determinants of glucocorticoid hormone action in the fetus with potential long-lasting consequences for stress adaptation and health.

Blunted serum and enhanced salivary free cortisol concentrations in the chronic phase after aneurysmal subarachnoid haemorrhage—is stress the culprit?

Spontaneous aneurysmal subarachnoid haemorrhage (SAH) is a cause of stroke, which constitutes a severe trauma to the brain and may lead to serious long-term medical, psychosocial and endocrinological sequelae. Adrenocorticotrophic hormone deficiency, which is considered to occur in up to 20% of all survivors, is a possible consequence of bleeding. Moreover, preliminary data suggest that a poor psychosocial outcome in SAH survivors is linked to alterations in cortisol secretion. Despite these findings, investigation of diurnal cortisol profiles and the cortisol awakening response (CAR) in chronic SAH patients has not been done so far. In this study, basal serum cortisol and salivary cortisol concentration profiles were investigated in 31 SAH patients more than 1 year after the acute event and in 25 healthy controls. Additionally, low-dose dexamethasone (DEX) suppression tests were conducted, and sensitivity to stress was measured with a psychometric questionnaire (NeuropatternTM). Although significantly higher salivary cortisol concentrations were observed on waking in SAH patients (p = 0.013, ANOVA), without a CAR change, total serum cortisol concentrations were blunted, but only in patients with high levels of perceived stress (SAH high stress: 337 nmol/l, SAH low stress: 442 nmol/l, controls: 467 nmol/l; Controls vs. SAH high stress p = 0.018). DEX suppression of cortisol secretion was not significantly different between patients and controls. The results indicate that total (serum) and free (salivary) cortisol concentrations give different information about cortisol availability in patients after aneurysmal SAH. Enhanced free cortisol concentrations may reflect a meaningful biological coping mechanism in SAH patients.

Resting Salivary Levels of IgA and Cortisol Are Significantly Affected During Intensive Resistance Training Periods in Elite Male Weightlifters

The aim of this study was to investigate the cumulative effects of intensive resistance training on salivary immunoglobulin A (SIgA) and cortisol responses in elite male weightlifters. Eleven elite male Taiwanese weightlifters were trained through 3 training stages before a national weightlifting competition, and this was followed by a 2-week recovery stage. Resting saliva samples were collected once in each of the 4 stages. Salivary concentrations of total protein (TP), SIgA, lactoferrin, and cortisol were measured. The results showed that (a) salivary TP concentrations were not significantly affected; (b) resting levels of SIgA, the ratio of SIgA to TP (SIgA/TP), cortisol, and the ratio of cortisol to TP (cortisol/TP) were significantly higher in the training stages than in the recovery stage; (c) a positive correlation was revealed between the ratios of SIgA/TP and cortisol/TP; and (d) the resting salivary lactoferrin concentrations and the ratio of lactoferrin to TP (lactoferrin/TP) were significantly lower in stage 1 than in the recovery stage. The findings in this study suggest that prolonged, intensive resistance training exerts cumulative effects on SIgA and cortisol responses in elite weightlifters.

Stability of the diurnal cortisol profile in children and adolescents

The diurnal cortisol profile has been implicated in multiple physical and mental health conditions in children and adolescents; however, current knowledge regarding the stability of the diurnal cortisol profile is largely based on adults. Developmental changes throughout childhood and adolescence warrant examination of the stability of the diurnal cortisol profile during this stage in the lifecourse. The aim of the present study was to conduct a comprehensive evaluation of the diurnal cortisol profile in children and adolescents. Participants (N=233; M=12.40, SD=1.83; 44.2% girls) in the Healthy Heart Project collected saliva samples, completed demographic questionnaires, and recorded bed and waking time. Intra-class correlations were calculated to evaluate the stability of aggregate and single sample measures of the diurnal cortisol profile. Total cortisol concentration (AUC(TG), AUC(AG)) and maximum sample were the most stable cortisol measures (ICC(avg)=0.54). Dynamic measures (AUC(I), slope; ICC(avg)=0.22) and other single sample measures (awake, lunch, dinner, bedtime, morning random, day random; ICC(avg)=0.28) were less stable. Of the developmentally relevant covariates tested, sleep duration, adrenarche, and time of awakening were most associated with cortisol values. Altogether, the diurnal cortisol profile yielded moderate to high stability in children and adolescents. These findings can inform methodological decisions regarding cortisol sampling protocols for children and adolescents.

Effects of 4:1 carbohydrate/protein solution versus a carbohydrate-alone solution on IL-6, TNF-&amp;alpha;, and cortisol during prolonged cycling in hot environmental conditions

PurposeIntense or prolonged exercise and/or heat stress might affect the immune system creating a response similar to trauma or inflammation, resulting in an increase in the susceptibility to viral infections. For example, during prolonged exercise, inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-6, and the stress hormone cortisol are produced and released. Although there have been several studies examining the effects of nutritional supplementation on cytokine release in elite athletes, few studies have investigated the effects of different energy drinks during exercise in adverse environmental conditions. Therefore, the purpose of this study was to compare plasma levels of inflammatory cytokines TNF-α and IL-6, and the stress hormone cortisol, during prolonged cycling under hot environmental conditions while ingesting fluid that contains a ratio of 4:1 carbohydrates and protein (4:1 CHO/PRO) versus a carbohydrate-only drink (CHO).MethodsSix male cyclists (aged 27 ± 8 years; weight 75.5 ± 3.4 kg; VO2max = 66 ± 2.7 mL/kg/min, mean ± standard error) rode on a stationary ergometer on two separate sessions for 2.5 hours at 75% VO2max in an environmental chamber set at 35°C and 60% relative humidity. During the first session the cyclists were given 4 mL/kg body weight of a 6% carbohydrate solution every 15 minutes. During the second session they were given 4 mL/kg body weight of a 4:1 carbohydrate/protein drink every 15 minutes. Subjects were not aware of which drink they were given in each trial. Blood samples were taken pre-, immediately post-, and 12 hours post-exercise. SPSS (IBM Corp, Armonk, NY) was utilized to analyze data through repeated measures analysis of variance.ResultsNo significant main effect was observed between treatments in either cortisol (P = 0.97), IL-6 (P = 0.64), or TNF-α (P = 0.37) responses. Total cortisol concentrations were significantly elevated (P < 0.05) immediately post-exercise, and from pre- to 12 hours post-exercise with both the 4:1 CHO/PRO and the CHO-alone solutions. TNF-α concentrations were only significantly (P = 0.045) elevated post-exercise with the CHO-alone solution. A significant (P < 0.05) elevation of IL-6 was seen immediately post-exercise and 12 hours post-exercise with both the CHO-alone and 4:1 CHO/PRO solutions.ConclusionsConsuming a 4:1 CHO/PRO solution during prolonged cycling under hot environmental conditions has comparable effects on inflammatory cytokines to drinking a CHO-alone solution.

English

&nbsp; This&nbsp;study&nbsp;was conducted to&nbsp;evaluate&nbsp;the effect of supplemental chromium on performance and blood serum biochemistry of dairy cows. Thus,&nbsp;20&nbsp;multiparous Holstein cows (parity 3) were equally divided into two groups, group one (control), which received no chromium supplementation and group two (treatment) which received 5 g/day chromium methionine from week 5 prior to parturition until 12 weeks thereafter. Milk production and milk composition were evaluated on 4, 8 and 12 weeks after parturition. Serum biochemistry concentrations (serum glucose, cholesterol, triglyceride, total protein, and cortisol and insulin concentration) and blood hematology (red blood cell, hematocrit, hemoglobin concentration and percentage neutrophils, lymphocytes, monocytes, basophiles, eosinophils and ratio of neutrophils to lymphocytes) were measured on 2 and 5 weeks prior to parturition and 1 and 4 weeks thereafter. Results indicate that milk production was significantly affected by chromium-methionine supplementation during the entire period (P&lt;0.01) but no significant effect on milk composition was found. Supplemental chromium had no significant effect on serum glucose, cholesterol, triglycerides and insulin concentration and blood hematology parameters (P&lt;0.05). However, chromium supplementation tended to increase significantly, serum total protein concentration and decrease cortisol concentration (P&lt;0.05). The results of this experiment showed that chromium methionine supplementation in multiparous dairy cows diet may improve their milk yield in transition period. &nbsp; Key words:&nbsp;Dairy cow, transition period, chromium-methionine, milk yield, serum biochemistry.

Free Cortisol and Accuracy of Total Cortisol Measurements in the Diagnosis of Adrenal Insufficiency in Brain-dead Patients

After brain death, adrenal insufficiency (AI) is very common and may be one of the mechanisms that contributes to hemodynamic instability and loss of potential organ donors. However, when diagnosed by total cortisol measurement, critically ill patients may be overdiagnosed as having AI. The aims of this study were to assess the prevalence of AI when diagnosed using free cortisol measurement and the accuracy of total cortisol measurement to diagnose AI in brain-dead patients. All consecutive brain-dead patients were included in this single-center noninterventional clinical observation study. Assessment of adrenocorticotropin, corticosteroid-binding globulin, baseline and tetracosactin-stimulated serum free and total cortisol concentrations were performed. AI was defined as a baseline free cortisol concentration ≤ 55 nM(-1) and/or Δ free cortisol ≤ 55 nM(-1). Patients were considered to have a low albumin concentration if less than 25 g · L(-1) and a low corticosteroid-binding globulin concentration if less than 27 mg · L(-1) in men or 31 mg · L(-1) in women. Among the 42 included patients, the incidence of AI was 83% (95% CI, 69-93%). Baseline total cortisol was correlated with baseline free cortisol, whatever the albumin or corticosteroid-binding globulin concentration. The area under the receiver operating characteristic curve of baseline total cortisol measurement to diagnose AI was 0.94 (95% CI, 0.81-0.98). The optimal cutoff was 485 nM(-1), providing a sensitivity and a specificity of 89% and 100%, respectively. Total baseline cortisol measurement is accurate and sufficient to diagnose AI in brain-dead patients, even if albumin or corticosteroid-binding globulin concentrations are low.

Coping with Intense Reproductive Aggression in Male Arctic Ground Squirrels: The Stress Axis and Its Signature Tell Divergent Stories

We tested the adaptive stress hypothesis that male arctic ground squirrels (Urocitellus parryii) exhibit a stress response over the course of the breeding season that is characterized by increasing free cortisol concentrations, increasing mobilization of stored energy, and decreasing physical condition. We assessed the functioning of the hypothalamic-pituitary-adrenal axis by measuring cortisol levels in response to the stress of capture and in response to a hormone challenge protocol (dexamethasone suppression and adrenocorticotropic hormone stimulation). We measured blood glucose levels, free fatty acids, white blood cells, and hematocrit to assess the downstream physiological responses to cortisol. Immediately after spring emergence, male arctic ground squirrels had ample free abdominal fat and few signs of wounding. By the end of the breeding season 3 wk later, visible fat reserves were almost entirely gone, and most males had extensive wounds. Total plasma cortisol concentrations increased over this period, but so did corticosteroid-binding capacity, resulting in no change in the free cortisol response to capture. We found no significant changes in how the animals responded to our hormone challenges, contrary to our prediction that the stress axis should increase free cortisol production. Even though we found no change in the functioning of the stress axis, all of the downstream measures suggested that male arctic ground squirrels are chronically exposed to high cortisol concentrations. Over the breeding season, blood glucose increased, fat stores and circulating free fatty acids were depleted, and both hematocrit levels and white blood cell counts decreased significantly. Our data suggest that a more complex relationship between the stress axis and downstream measures of stress exists than that proposed by the adaptive stress hypothesis. We propose several nonexclusive, testable mechanisms that could explain our observations.

Predictors of neonatal hypothalamic-pituitary-adrenal axis activity at delivery.

Clinical and preclinical studies indicate that maternal stress during pregnancy may exert long-lasting adverse effects on offspring. This investigation sought to identify factors mediating the relationship between maternal and neonatal hypothalamic-pituitary-adrenal (HPA) axes in pregnant women with past or family psychiatric history. Two hundred and five pairs of maternal and umbilical cord blood samples from a clinical population were collected at delivery. Maternal and neonatal HPA axis activity measures were plasma adrenocorticotrophic hormone (ACTH), total cortisol, free cortisol and cortisol-binding globulin concentrations. The effects of maternal race, age, body mass index, psychiatric diagnosis (DSM-IV), birth weight, delivery method and estimated gestational age (EGA) at delivery on both maternal and neonatal HPA axis measures were also examined. Incorporating these independent predictors as covariates where necessary, we evaluated whether neonatal HPA axis activity measures could be predicted by the same maternal measure using linear regression. Delivery method was associated with umbilical cord plasma ACTH and both total and free cord cortisol concentrations (T = 10·53-4·21; P < 0·0001-0·010). After accounting for method of delivery and EGA, we found that maternal plasma ACTH concentrations predicted 23·9% of the variance in foetal plasma ACTH concentrations (T = 6·76; P < 0·0001), and maternal free and total plasma cortisol concentrations predicted 39·8% and 32·3% of the variance in foetal plasma free and total cortisol concentrations (T = 5·37-6·90; P < 0·0001), respectively. These data suggest that neonatal response is coupled with maternal HPA axis activity at delivery. Future investigations will scrutinize the potential long-term sequelae for the offspring.