Higher Plasma Cortisol Levels Research Articles

Sea water acclimation of rainbow trout (Oncorhynchus mykiss) modulates the mucosal transcript immune response induced by Vibrio anguillarum and Aeromonas salmonicida vaccine, and prevents further transcription of stress-immune genes in response to acute stress

Mucosal tissues appear to be more important in fish than in mammals due to living in a microbial-rich aquatic milieu, yet the complex interaction between the immune and the neuroendocrine system in these tissues remains elusive. The aim of this work was to investigate the mucosal immune response in immunized rainbow trout vaccinated with Alpha ject vaccine (bivalent), kept in fresh water (FW) or transferred to seawater (SW), and to evaluate their response to acute stress (chasing). Acute stress resulted in higher levels of plasma cortisol (Sham + Stress and Vaccine + Stress). A similar response was observed in skin mucus, but it was lower in Vaccine + Stress compared with stressed fish. With a few exceptions, minimal alterations were detected in the transcriptomic profile of stress-immune gene in the skin of vaccinated and stressed fish in both FW and SW. In the gills, the stress elicited activation of key stress-immune components (gr1, mr, β-ar, hsp70, c3, lysozyme, α-enolase, nadph oxidase, il1β, il6, tnfα, il10 and tgfβ1) in FW, but fewer immune changes were induced by the vaccine (nadph oxidase, il6, tnfα, il10 and igt) in both SW and FW. In the intestine, an array of immune genes was activated by the vaccine particularly those related with B cells (igm, igt) and T cells (cd8α) in FW with no stimulation observed in SW. Therefore, our survey on the transcriptomic mucosal response demonstrates that the immune protection conferred by the vaccine to the intestine is modulated in SW. Overall, our results showed: i) plasma and skin mucus cortisol showed no additional stress effect induced by prolonged SW acclimation, ii) the stress and immune response were different among mucosal tissues which indicates a tissue-specific response to specific antigens/stressor. Further, the results suggest that the systemic immune organs may be more implicated in infectious events in SW (as few changes were observed in the mucosal barriers of immunized fish in SW) than in FW.

O-03 Adrenalectomy for Cushing's syndrome during pregnancy

Abstract Introduction Cushing's syndrome (CS) is rare in pregnancy because CS itself is a cause of anovulatory infertility due to excess cortisol secretion. Moreover it is also difficult to diagnose CS in pregnancy because placental production of corticotropin releasing and adrenocorticotropic hormone (ACTH), increased maternal production of estrogen induced corticosteroid binding globulin leads to physiological hypercortisolism during pregnancy. So physicians face a clinical overlap in many features of CS such as weight gain, fatigue, edema, hypertension, hyperglycemia, stretch marks, mood changes that can also be attributed to pregnancy. Thus their coexistence represents a diagnostic challenge. Herein we report a case of CS that was diagnosed and successfully treated during pregnancy. Clinical Case A 43-year-old gravida 3 female was consulted at the 21st week of gestation with persistent high blood pressure and hyperglycemia for a month. She had an obstetric history of 2 spontaneous first term abortion, 2 healthy labor 18 years ago and she was healthy after. She was in such a depressive mood that she was consulted with psychiatry many times. On physical examination edema, proximal weakness of lower extremities and hirsutism (modified Ferriman–Gallwey score of 9) was detected. Initial laboratory tests were all within normal range except potassium level (2.7 mmol/L), the results were not consistent with preeclampsia and there was a suspicious adrenal mass detected on ultrasound. She was currently receiving high dose antihypertensive treatment with methyldopa, calcium channel blockers, rapid acting and detemir insulin for diabetes and i.v. potassium replacement. We ruled out conns syndrome with normal plasma aldosterone renin ratio and found catecholamine levels normal. Hormonal analyses revealed with high plasma cortisol levels, increased nocturnal cortisolemia, a suppressed plasma ACTH and dehydroepiandrosterone sulfate level and distinctly elevated 24-hour urinary cortisol greater than 3-fold the upper normal reference range. Non contrast magnetic resonance imaging demonstrated a 47×32 mm left adrenal mass. Laparoscopic adrenalectomy was performed in the 26th week of gestation and the following histopathology revealed an adrenocortical adenoma. Antihypertensive medication was progressively reduced, glycemic control and hypokalemia reversal were achieved. She was followed up with appropriate glucocorticoid replacement. At 34 weeks of gestation, after sudden labor pain and regular uterine contractions a 2586 g male infant with Apgar scores of 8 was born by cesarean section. They were discharged in good health. Conclusion CS should be kept in mind especially in the case of hypertension and diabetes that started before the 20th gestational age. Analysis of the urinary free cortisol level and circadian rhythm of cortisol is necessary to diagnose. Despite the diagnostic challenge we can achieve better maternal and fetal outcomes with proper management.Figure 1.Non-contrast magnetic resonance imaging of the patient Table 1.Patient's laboratuary test results Table 2.Patient's laboratory test results

Plasma cortisol-linked gene networks in hepatic and adipose tissues implicate corticosteroid-binding globulin in modulating tissue glucocorticoid action and cardiovascular risk.

Genome-wide association meta-analysis (GWAMA) by the Cortisol Network (CORNET) consortium identified genetic variants spanning the SERPINA6/SERPINA1 locus on chromosome 14 associated with morning plasma cortisol, cardiovascular disease (CVD), and SERPINA6 mRNA expression encoding corticosteroid-binding globulin (CBG) in the liver. These and other findings indicate that higher plasma cortisol levels are causally associated with CVD; however, the mechanisms by which variations in CBG lead to CVD are undetermined. Using genomic and transcriptomic data from The Stockholm Tartu Atherosclerosis Reverse Networks Engineering Task (STARNET) study, we identified plasma cortisol-linked single-nucleotide polymorphisms (SNPs) that are trans-associated with genes from seven different vascular and metabolic tissues, finding the highest representation of trans-genes in the liver, subcutaneous fat, and visceral abdominal fat, [false discovery rate (FDR) = 15%]. We identified a subset of cortisol-associated trans-genes that are putatively regulated by the glucocorticoid receptor (GR), the primary transcription factor activated by cortisol. Using causal inference, we identified GR-regulated trans-genes that are responsible for the regulation of tissue-specific gene networks. Cis-expression Quantitative Trait Loci (eQTLs) were used as genetic instruments for identification of pairwise causal relationships from which gene networks could be reconstructed. Gene networks were identified in the liver, subcutaneous fat, and visceral abdominal fat, including a high confidence gene network specific to subcutaneous adipose (FDR = 10%) under the regulation of the interferon regulatory transcription factor, IRF2. These data identify a plausible pathway through which variation in the liver CBG production perturbs cortisol-regulated gene networks in peripheral tissues and thereby promote CVD.

Transcutaneous auricular vagus nerve stimulation may elicit anti-inflammatory actions through activation of the hypothalamic-pituitary-adrenal axis in humans

Introduction: Since the discovery of the cholinergic anti-inflammatory pathway, vagus nerve stimulation (VNS) has been suggested as a treatment option for chronic inflammatory conditions. This pathway relies on efferent vagal nerve fibers. Invasive cervical VNS activates afferent and efferent vagal nerve fibers and, thus, has the potential to directly activate the cholinergic anti-inflammatory pathway. In contrast, transcutaneous auricular VNS (taVNS) activates the auricular branch of the vagus nerve that is purely afferent and projects to the nucleus of the solitary tract. Thus, any anti-inflammatory actions of taVNS are unlikely to result from a direct activation of the cholinergic anti-inflammatory pathway. The objective of this study was to investigate alternative mechanisms by which taVNS may elicit anti-inflammatory effects. The hypothesis was tested that taVNS activates the hypothalamic-pituitary-adrenal (HPA) axis, resulting in adrenal cortisol secretion that will then modulate circulatory immune cell numbers. Methods: The study was approved by the appropriate authorities (IRB# 0054_2019 and NCT04177264). Study participants were healthy adults (n=19, 6 male, 13 female). On three consecutive days either taVNS (left ear, 10 Hz, 1-2 mA, 300 μs pulse width) or sham-taVNS (stimulator off) was applied for 10 minutes. A blood sample was drawn at the end of the third day. Plasma cortisol was determined by ELISA (K003-H1W, Arbor Assays, MI). Circulating T-helper cells (CD3+, CD4+), cytotoxic T-cells (CD3+, CD8+), B-cells (CD19+), monocytes (CD14+, CD11b+), and natural killer (NK, CD3-, CD56+) cells were counted using flow cytometry. Statistical significance was assumed at P<0.05 and trends are reported for P<0.15 based on unpaired t-tests. Results: Plasma cortisol levels tended to be higher following taVNS (72.5±9.5 ng/mL) compared to sham-taVNS (52.3±3.8 ng/mL, P=0.09). Following taVNS, the relative number of circulatory B cells was lower (0.48±0.11 % vs. 0.82±0.11 %, P<0.05), while the number of circulatory monocytes was higher (2.21±0.32 % vs 1.58±0.16 %, P=0.06) compared to sham-taVNS. T-helper cells, cytotoxic T cells, and NK cells were not significantly affected by taVNS. In conclusion, the higher plasma cortisol levels following taVNS together with the higher number of circulatory monocytes and lower number of B cells suggest that taVNS elicits anti-inflammatory actions through activation of the HPA axis, because it is known that cortisol increases circulatory monocytes and reduces B cells. The decrease in B cells following taVNS gives rise to the intriguing hypothesis that taVNS may prevent secondary treatment failure to biologic drugs in chronic inflammatory conditions by reducing production of anti-drug antibodies. Funding: This study was supported by a grant from the American Osteopathic Association (Grant No.: 19137759). This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

PS-R03-3: INTERMITTENTLY SCANNED CONTINUOUS GLUCOSE MONITORING IN A DIFFICULT PATIENT WITH GESTATIONAL DIABETES MELLITUS AND SUBCLINICAL CUSHING'S SYNDROME: A CASE REPORT

A 41-year-old woman was admitted to the obstetrics department because of epigastric pain, nausea, and fatigue at 31 weeks of gestation. She was suspected of subclinical Cushing's syndrome based on high plasma cortisol and low ACTH levels along with an incidentally discovered tumor in the left adrenal gland 4 years prior to conception. 3 months before admission, 75 g oral glucose tolerance test yielded one point positive (81–169–174 mg/dL) and diagnosis of gestational diabetes mellitus was made. Diet and exercise therapy was started on an outpatient basis. While blood glucose control had been favorable, she discontinued visits to outpatient clinics due to difficulty in physical activity. At the time of admission, blood glucose level was running more than 200 mg/dl, and self-monitoring of blood glucose (SMBG) and insulin self-injection therapy were proposed for immediate blood glucose control accordingly. However, the patient had psychiatric symptoms that were presumably associated with delusional disorder and subclinical Cushing's syndrome. As such, she refused to implement both procedures on account of a strong fear against puncture, which caused blood glucose management difficult. Despite understanding the necessity of SMBG and potential risks to the fetus, the patient frequently expressed anxiety and could not perform SMBG. Given the difficult circumstance, intermittently scanned continuous glucose monitoring was installed to reduce invasive procedures. As a result, blood glucose levels were under good control with 0.43 U/kg of insulin per day until the delivery by cesarean section at 34 weeks of gestation. A healthy infant weighing 2442 g was delivered without complications. Here we highlight utility and efficacy of intermittently scanned continuous glucose monitoring in a difficult patient with gestational diabetes mellitus and subclinical Cushing's syndrome.

The developmental consequences of early adverse care on infant macaques: A cross-fostering study

Early life adversity/stress (ELA/ELS), particularly adverse caregiving experiences such as child maltreatment (MALT), is a main risk factor for psychopathology, including psychiatric disorders such as anxiety, depression, ADHD, and substance abuse. Yet how these alterations unfold during development and the underlying mechanisms remain poorly understood, as it is difficult to prospectively and longitudinally study early developmental phases in humans, and nearly impossible to disentangle postnatal caregiving effects from heritable traits. This study examined the specific effects of “nurture” (maternal care) versus “nature” (heritable, biological maternal factors) on nonhuman primate infant socioemotional, stress neuroendocrine, and physical development. For this we used a translational and naturalistic macaque model of infant maltreatment by the mother with randomized assignment at birth to either mothers with a history of maltreating their infants (MALT group, n = 22) or to competent mothers (Control group, n = 20). Over the first 6 months of life (roughly equivalent to 2 years in humans), we examined the development of the mother-infant relationship, as well as infants’ social behavior and emotional reactivity. In parallel, we assessed hypothalamic-pituitary-adrenal (HPA) axis function longitudinally, using measures of hair cortisol accumulation, and basal morning plasma cortisol. We identified broad impairments in maternal care exhibited by MALT foster mothers, beyond maltreatment (physical abuse, rejection) events, suggesting that MALT foster mothers provide an overall lower quality of care to their infants compared to Controls. MALT infants exhibited alterations in their initiations and breaks of proximity towards their mothers, as well as heightened emotional reactivity in comparison to Controls. Most striking are the HPA axis findings, with MALT infants showing higher levels of plasma cortisol across the first 6 postnatal months as well as higher hair cortisol accumulation from birth through month 6 (a signature of chronic stress) than Controls. No caregiving effects were detected on physical growth, which ruled out confounding effects of maternal nutrition, metabolism, etc. Taken together, these results suggest that the developmental trajectory of MALT and Control infants is different, marked by heightened levels of emotional reactivity, increased HPA activity and alterations in mother-infant interactions in MALT animals. These findings appear to be due to specific effects of postnatal maternal care, and not to biological/ behavioral traits inherited from the mother, or due to prenatal programming caused by prenatal stress, as the cross-fostering design controlled for these potential factors. However, we also detected a couple of interesting biological effects suggesting heritable transmission of some phenotypes. The prolonged HPA axis activation during the first 6 postnatal months of life is expected to have long-term consequences for brain, physiological, and behavioral development in MALT offspring.

Association of Plasma Cortisol Levels with Gestational Age and Anthropometric Values at Birth in Preterm Infants.

There are no study reports to clarify the association between gestational age (GA) or anthropometric values at birth, and plasma cortisol levels in the blood of preterm infants at birth and at one month of age. This hospital-based retrospective cohort study included infants born at <37 weeks’ gestation between 2019 and 2021. First, the association between plasma cortisol level and GA or anthropometric values at birth (birth weight standard deviation score [SDS], birth length SDS, and birth head circumference SDS) was identified by regression and multiple regression analyses. Second, plasma cortisol levels in the umbilical cord at birth and at one month of age were compared between small-for-gestational age (SGA) and non-SGA infants. Sixty-one preterm infants were enrolled (SGA: 24 and non-SGA: 37). Plasma cortisol levels at birth were significantly associated with GA. Plasma cortisol levels at one month of age were associated with GA and birth head circumference SDS. Plasma cortisol levels at birth were significantly higher in SGA than non-SGA (p = 0.010). GA was an independent determinant of plasma cortisol levels at birth. SGA infants had a high plasma cortisol level at birth; resulting in speculation that a high plasma cortisol level at birth may predict abnormal neurological outcomes.

Metabolic scope, performance and tolerance of juvenile European sea bass Dicentrarchus labrax upon acclimation to high temperatures.

European sea bass is a species of great commercial value for fisheries and aquaculture. Rising temperatures may jeopardize the performance and survival of the species across its distribution and farming range, making the investigation of its thermal responses highly relevant. In this article, the metabolic scope, performance, and tolerance of juvenile E. sea bass reared under three high water temperatures (24, 28, 33°C), for a period of three months was evaluated via analysis of selected growth performance and physiological indicators. Effects on molecular, hormonal, and biochemical variables were analyzed along with effects of acclimation temperature on the metabolic rate and Critical Thermal maximum (CTmax). Despite signs of thermal stress at 28°C indicated by high plasma cortisol and lactate levels as well as the upregulation of genes coding for Heat Shock Proteins (HSP), E. sea bass can maintain high performance at that temperature which is encouraging for the species culture in the context of a warming ocean. Critical survivability thresholds appear sharply close to 33°C, where the aerobic capacity declines and the overall performance diminishes. European sea bass demonstrates appreciable capacity to cope with acute thermal stress exhibiting CTmax as high as 40°C for fish acclimated at high temperatures, which may indicate resilience to future heatwaves events.

How distinct killifish populations respond differently to stress

Down the east coast of North America, populations of Atlantic killifish, Fundulus heteroclitus, experience wildly different thermal environments. Southern populations experience water that is, on average, 12°C hotter than northern populations annually. These northern and southern killifish populations cope with stress differently. When fish are stressed, such as when predators are nearby, they activate a suite of mechanisms that regulate the release of the stress hormone cortisol, allowing the fish to remain in good condition. Researchers found previously that killifish from southern populations had higher plasma cortisol levels than killifish from northern populations after they experienced the same stress. To figure out why some fish respond strongly to stress while others do not, Madison Earhart and co-workers from the University of British Columbia, Canada, with colleagues from the University of Manitoba, Canada, and the University of Glasgow, UK, set out to determine what genes are responsible for the differences in killifish stress responses by investigating different mechanisms that trigger cortisol release after experiencing stress.Earhart and colleagues travelled to the USA and collected adult killifish from northern New Hampshire and southern Georgia populations. Then, they brought the fish back to the lab in British Columbia. First, the team wanted to see whether the differences in population stress response changed when the fish experienced stress briefly or were exposed to the same stress repeatedly over the period of a week by either putting the fish in buckets and shaking them for half an hour or repeatedly shaking them every day for a week. After stressing the fish, the team collected samples of the fish's blood to measure their cortisol plasma levels, in addition to collecting samples of the fish's brain, head kidney – the organ that produces cortisol – and liver – the organ that produces and breaks down glucose – to measure the expression of genes important for the production and release of cortisol due to stress.After experiencing the single stressful situation, the southern killifish population had higher cortisol levels than the northern killifish population. However, when the fish were repeatedly stressed over the period of a week, there was no difference in cortisol levels between the northern and southern populations. The southern population of killifish, which had a stronger response to the individual stressful situation, expressed more of the genes involved in cortisol production in the brain and head kidney. In addition, the livers of fish from the southern population were more responsive to the cortisol stress hormone as they expressed larger amounts of the gene for the protein that triggers the protective mechanisms that are activated by cortisol when a fish is stressed.The team also measured the condition of the fish after repeated stress. They found that the southern killifish, which were more stress responsive than the northern killifish, were in better condition after experiencing repeated stress than were northern populations. This showed that responding to stress by increasing cortisol was beneficial for the southern population of fish.Earhart and colleagues noted that the answer as to why some fish respond more strongly to stress than others is more complex than simply increasing cortisol levels in the body. They showed that there are differences in the expression of genes all along the pathways that make cortisol, respond to cortisol and cease cortisol production in these two different populations of fish. Therefore, it is important to continue to study these differences in populations of the same species adapted to different environments.

Review of Mendelian Randomization Studies on Endometrial Cancer.

Endometrial cancer (EC) is a common gynecological cancer. In some parts of the world, the incidence and mortality of EC are on the rise. Understanding the risk factors of EC is necessary to prevent the occurrence of this disease. Observational studies have revealed the association between certain modifiable environmental risk factors and EC risk. However, due to unmeasured confounding, measurement errors, and reverse causality, observational studies sometimes have limited ability to judge robust causal inferences. In recent years, Mendelian randomization (MR) analysis has received extensive attention, providing valuable insights for cancer-related research, and is expected to identify potential therapeutic interventions. In MR analysis, genetic variation (alleles are randomly assigned during meiosis and are usually independent of environmental or lifestyle factors) is used instead of modifiable exposure to study the relationship between risk factors and disease. Therefore, MR analysis can make causal inference about exposure and disease risk. This review briefly describes the key principles and assumptions of MR analysis; summarizes published MR studies on EC; focuses on the correlation between different risk factors and EC risks; and discusses the application of MR methods in EC research. The results of MR studies on EC showed that type 2 diabetes, uterine fibroids, higher body mass index, higher plasminogen activator inhibitor-1 (PAI-1), higher fasting insulin, early insulin secretion, longer telomere length, higher testosterone and higher plasma cortisol levels are associated with increased risk of EC. In contrast, later age of menarche, higher circulatory tumor necrosis factor, higher low-density lipoprotein cholesterol, and higher sex hormone-binding globulin levels are associated with reduced risk of EC. In general, despite some limitations, MR analysis still provides an effective way to explore the causal relationship between different risk factors and EC.

Paraneoplastisch syndroom van Cushing

Paraneoplastic Cushing’s syndrome Introduction. Paraneoplastic Cushing’s syndrome (ECS) is a rare condition which is related to an ectopic adrenocorticotropic hormone (ACTH) or, rarely, corticotropin-releasing hormone (CRH) production. Diverse tumours can cause ECS; small-cell lung cancer (SCLC) and bronchial carcinoids account for the majority of the cases. In this report, the medical history of a patient with ECS due to SCLC is presented. The main purpose of this case-report is to outline the difficult clinical presentation. Case description. A 72-year-old cachectic man was diagnosed with SCLC. The coexistence of hypokalemia and hyperglycemia raised the suspicion of ECS, which was confirmed by high levels of plasma cortisol, plasma ACTH and urinary cortisol. Discussion. The diagnostic process of Cushing’s syndrome is outlined. Conclusion. The diagnosis of ECS can be challenging since typical clinical symptoms may be absent. Hypokalemia or hyperglycemia in a context of SCLC or a neuro-endocrine tumour (NET) should prompt further evaluation with determination of the plasma cortisol and ACTH.

Predisposing factors for increased cortisol levels in oral cancer patients

Cancer patients may have a dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis and abnormal secretion of cortisol. Increased cortisol levels have been associated with worse prognosis in patients with different types of tumors. Although anxiety and depression can trigger an abnormal cortisol secretion, little is known regarding the influence of these emotional disorders on HPA axis dysregulation in cancer patients when evaluating together with demographic, clinicopathological and biobehavioral variables. This cross-sectional study analyzed the pre-treatment plasma cortisol levels of 133 patients with oral squamous cell carcinoma (OSCC) and its association with demographic, clinicopathological, biobehavioral and psychological variables. Plasma cortisol levels were measured by electrochemiluminescence, and anxiety and depression symptoms were assessed using Beck Anxiety Inventory (BAI) and Depression (BDI), respectively. Demographic, clinicopathological and biobehavioral data were collected from patients' medical records. Results from multivariate analysis showed that the occurrence of cancer-induced pain was predictive for higher cortisol levels (OR = 5.388, p = 0.003). Men with OSCC were 4.5 times more likely to have higher plasma cortisol levels than women (OR = 4.472, p = 0.018). The effect of sex on cortisol concentrations was lost in the adjusted model for clinical staging (OR = 2.945, p = 0.116). The absence of chronic alcohol consumption history was a protective factor for highest hormone concentrations in oral cancer patients (OR = 0.104, p = 0.004). Anxiety symptoms measured by BAI as “hands trembling” (OR = 0.192, p = 0.016) and being “nervous” (OR = 0.207, p = 0.0004) were associated with lower cortisol levels. In contrast, the feeling of “fear of losing control” was a risk factor for highest hormone concentrations (OR = 6.508, p = 0.0004). The global score and specific symptoms of depression measured by the BDI were not predictive for plasma hormone levels (p > 0.05). Together, our results show that pain, alcohol consumption and feeling fear are independent factors for increased systemic cortisol levels in patients with oral cancer. Therefore, psychological intervention, as well as control of pain and alcohol consumption, should be considered to prevent the negative effects of cortisol secretion dysregulation in cancer patients.

Changes in the pituitary-adrenal system for extracorporal membrane oxygenation: prospective study

INTRODUCTION. At the moment, assessing adrenal dysfunction in critical patients and ways to correct this function with hormone replacement therapy are extremely difficult. OBJECTIVES. Analysis of changes in the dynamics of ACTH and cortisol levels in blood plasma during ECMO. MATERIALS AND METHODS. The prospective study was performed in intensive care unit (47 patients on ECMO). After connecting ECMO (D0), (D1–D3–D5–D7–D9), and until the completion of ECMO, assessment of cortisol and ACTH levels was carried out. RESULTS. The median level of cortisol in blood plasma was higher in the non-survivors patients on the third day (D3) (p = 0.05), D7 (p = 0.03); D13 (p = 0.05) and the last day of observation (p = 0.001), respectively. The level of ACTH in the blood of non-survivors patients was higher immediately on the day of ECMO initiation (D0) and on day 3 (D3) of observation: D0 (p = 0.018); D3 (p = 0.04), respectively. Analysis of the ROC curve showed that cortisol levels show a sensitivity of 71 % and a specificity of 89 % to an adverse outcome during ECMO. DISCUSSION. The life-saving ECMO technique, in critical conditions, is associated with a high risk of increasing of complications, including potentially lethal ones. Critical illness-related corticosteroid insufficiency (CIRCI) clinically manifests itself as inadequate adrenal activity, taking into account the augmentation of the disease severity. This activity is expressed in the form of a decrease in production and/or resistance to endogenous cortisol, as confirmed by the study. Consideration of CIRCI during the usage of ECMO reflects more objectively the violation of the pituitary-adrenal system. CONCLUSIONS. 1. CIRCI is detected in patients during ECMO. 2. High plasma cortisol levels are the predictor of an adverse outcome. 3. The level of ACTH in blood plasma is higher in patients with adverse outcomes. 4. High levels of cortisol in plasma are not a criterion for making the decision to initiate hydrocortisone therapy.

Social effects on AVT and CRF systems

Stress and aggression have negative effects on fish welfare and productivity in aquaculture. Thus, research to understand aggression and stress in farmed fish is required. The neuropeptides arginine-vasotocin (AVT) and corticotropin-releasing factor (CRF) are involved in the control of stress and aggression. Therefore, we investigated the effect of agonistic interactions on the gene expression of AVT, CRF and their receptors in juvenile rainbow trout (Oncorhynchus mykiss). The social interactions lead to a clear dominant-subordinate relationship with dominant fish feeding more and being more aggressive. Subordinate fish had an upregulation of the AVT receptor (AVT-R), an upregulation of CRF mRNA levels, and higher plasma cortisol levels. The attenuating effect of AVT on aggression in rainbow trout is proposed to be mediated by AVT-R, and the attenuating effect of the CRF system is proposed to be mediated by CRF.

Mismothering and remedying the mother-young relationship in stabled dromedary camels

This study aimed to characterize inappropriate maternal behaviours in stabled dromedary camels in order to detect the main causes of calf rejection, and to develop a suitable method to remediate mismothering. A herd of 45 pregnant females was under video surveillance to detect periparturient females. Twelve pregnant dromedary females (6 accepting their newborns and 6 rejecting them 1 h after calving) were selected as experimental subjects. A blood sample was collected during calving to determine cortisol levels. Maternal behaviour at the calving site was assessed during the first hour postpartum. Rejecting dams and their neonates were moved into a confinement enclosure to assess their maternal behaviour over a period of 30 min at 1 h, 6 h, 12 h, 24 h, 48 h, 72 h during confinement and at 7 d postpartum. Five IU of synthetic oxytocin were injected at 8:00, 12:00 and 16:00 to induce milk ejection in rejecting dams and one qualified person collected colostrum. The newborns were fed by holding the bottle of colostrum on the surface of the mother’s udder and keeping body contact between the two animals. Results showed that the duration of parturition was longer in the mothers that rejected their calves (P = 0.04). Such females spent more time in standing position during the calving process (P = 0.01) compared to accepting mothers, had higher plasma cortisol levels (P < 0.01), and gave birth preferentially while standing. Rejecting mothers changed calving sites and displayed head-turning more frequently (P < 0.01 in both parameters) than accepting mothers. Two causes of rejection were identified: dystocia and presence of an alien calf at the birth site. Moreover, a female showed a case of repeated rejecters. During the first hour postpartum, rejecting mothers failed to express good maternal care and moved away from their neonates. In the confinement enclosure, they exhibited attack behaviour toward their calves, displayed low sniffing activity, and emitted mainly high-pitched grunt. Aggression diminished progressively and disappeared (P < 0.0001) on the 2nd day of confinement while sniffing behaviour increased steadily (P = 0.001). The number of high-pitched grunt decreased over time while the number of low-pitched grunt increased over the first 3 days before declining significantly at the end of the confinement period (P < 0.0001). In conclusion, rejecting mothers can be very aggressive towards their own newborns. However, three days of confinement and forced contact with the calf while bottle-feeding it with the mother’s milk was sufficient to alleviate mismothering in dromedaries and establish proper mother-young relationships.

Linking coping styles and enzymatic activity of energy metabolism in the wedge sole (Dicologlossa cuneata)

Knowledge of individual behavioural variations (coping styles) can be a very useful tool for aquaculture, since it could allow to adapt the culture conditions to the different behavioural traits and, thus, to reduce the stress in the fish getting in this way a better production. The objective of this work was the identification of coping styles and their possible association with energy metabolism enzyme activities in the wedge sole (Dicologlossa cuneata). After submitting the fish to different tests (Feeding Recovery, Novel Object, Aggression and Restraining), the results showed a high individual variation, especially in the adaptation to a new environment (CV = 90.92 %), latency to escape (CV = 114.14 %) and plasma cortisol levels (CV = 105.56 %), thus showing the existence of coping styles in this species. Two different personalities were defined: the CS1 group (reactive individuals) and the CS2 group (proactive individuals). The CS1, unlike the CS2, presented a shorter latency to escape, higher number of escape attempts, a longer duration of escape attempts, and higher plasma cortisol levels after an acute stress. In addition, significant differences were detected for Glycogen phosphorylase (GPase) that acts in glycogenolysis, presenting the group CS1 the highest values (0.80 ± 0.17 vs. 0.48 ± 0.02 U mg prot−1). In conclusion, this study demonstrates for the first time the existence of coping style in D. cuneata, proving effective the tests applied for this identification. Two different groups were observed with specific behavioural and physiological responses that fit the proactive and reactive patterns. Furthermore, these groups showed differences in the energy metabolism of carbohydrates in basal state.

Involvement of purinergic signaling in the Amazon fish Pterygoplichthys pardalis subjected to handling stress: Relationship with immune response

Stress has a considerable impact on the welfare and productivity of fish, and stress caused by transport and handling can result in mortality rates of up to 73%. Cortisol is the major corticosteroid produced by the hypothalamic-pituitary-interrenal axis (HPI) in several stressful conditions, and high plasma cortisol levels impair immune and inflammatory responses. However, the involvement of purinergic signaling, a system that might contribute to the fine tuning of inflammatory and immune responses caused by the stress response, remains unknown in fish subjected to handling stress. Thus, the aim of this study was to evaluate the involvement of purinergic signaling with the immune response of the Amazon fish bodó (Pterygoplichthys pardalis) to acute handling stress. Plasma cortisol levels were higher 240 min post-stress in fish subjected to handling than in the control group. Plasma triphosphate diphosphohydrolase (NTPDase) activity, using adenosine triphosphate (ATP) as a substrate, was upregulated 30 min post-stress in fish subjected to handling, whereas splenic NTPDase activity was downregulated 240 min post-stress compared to the control group. No significant difference was observed between the groups regarding plasma and spleen NTPDase (using adenosine diphosphate as substrate) and 5′-nucleotidase and adenosine deaminase (ADA) activities. The levels of nitric oxide metabolites (NOx) and ATP in the spleen were higher 240 min post-stress in fish subjected to handling, although no significant difference was observed in the plasma levels. Based on these results, purinergic signaling via regulation of extracellular ATP catabolism could be involved in the immunophysiological responses of bodó subjected to acute handling stress, exerting two different profiles. In plasma, this signaling contributes to a reduction in the systemic inflammatory response, however, inhibition of ATP hydrolysis in the spleen displayed a pro-inflammatory profile due to the excessive ATP levels in the extracellular milieu, which induced the release of pro-inflammatory mediators. Thus, NTPDase activity could be a pathway associated with the immunophysiological responses of bodó to acute handling stress.

Plasma cortisol is associated with cerebral hypometabolism across the Alzheimer's disease spectrum

Hypothalamic-pituitary-adrenal dysregulation is proposed as a risk factor for Alzheimer's disease (AD). This study assessed cross-sectional relationships between cortisol and neuroimaging biomarkers of brain structure and glucose metabolism across the AD spectrum. Participants with normal cognition, mild cognitive impairment, and AD were selected from the Alzheimer's Disease Neuroimaging Initiative databank, based on baseline measures of plasma cortisol, gray matter volume (n = 556), and cerebral glucose metabolism (n = 288). Relationships between plasma cortisol and the neuroimaging biomarkers were assessed. Across the entire cohort, higher plasma cortisol levels were associated with lower glucose metabolism in lateral and medial parietal regions. Higher plasma cortisol was also related to lower gray matter volume in temporal-parietal-occipital regions and in the hippocampus. There were no significant group differences in these relationships with adjustment for covariates. Our results demonstrate that hypothalamic-pituitary-adrenal axis activation is related to glucose hypometabolism within posterior cortical regions vulnerable to AD pathology. This regional pattern appears to be distinct from cortisol-related associations with brain structure. Future studies should delineate pathophysiological mechanisms underlying these effects.

CARDIOVASCULAR RISK AND CORTISOL SECRETION IN ADRENAL INCIDENTALOMAS

Objective: Adrenal incidentalomas (AIs) are often discovered after an imaging-work up that is unrelated to specific adrenal gland disease. Up to 30% of AIs show endogenous cortisol excess, without signs or symptoms of clinically evident Cushing's syndrome. According to literature, subclinical hypercortisolism (SH) is more frequently associated to hypertension as well as obesity and metabolic disorders. Previous studies have established in SH higher cardiovascular (CV) risk, and increased incidence of CV events. Aim of the study was to confirm the existence of a linear relationship between augmented level of cortisol obtained by 24h-urinary free cortisol and overnight 1-mg dexamethason suppression test, and worsening clinical outcomes.Design and method: We retrospectively analyzed data from 543 patients (224 F, 319 M; mean age 59.6+12.4 yrs) with AIs, referred to our Unit between January 2000 to December 2017. After follow-up duration of at least 36 months, we have registered main comorbidities, such as cardiovascular events (pulmonary embolism, deep vein thrombosis, myocardial infarction), metabolic disorders, neoplasms and autoimmune disorders. Results: In Table 1 we reported anthropometric and laboratory data baseline in patients with non-producing - AIs and SH patients. 33 patients with SH (13 M, 20 F; mean age 58.44+10,76 yrs) underwent unilateral laparoscopic adrenalectomy; 48 patients (23 M, 25 F; mean age 57.21+8 yrs) performed a conservative pharmacological treatment. Surgery group showed a minor prevalence of cardiovascular events than conservative group, especially acute myocardial infarction and related therapeutic procedures (percutaneous stenting and surgical bypass) (0.03 % and 0.03 % vs 11 % and 8.9 %, respectively). Conclusions: In SH patients higher levels of 24h-urinary free cortisol and higher levels of plasma cortisol after overnight 1-mg dexamethason suppression test are strictly linked to higher levels of total cholesterol, LDL cholesterol and uric acid, as well as worsen clinical outcomes. Our results confirm that hormonal activity and autonomous production of cortisol is related to higher prevalence of cardiovascular diseases, i.e. arterial hypertension, and CV complications, as myocardial infarction.

Neuropsychological assessment and cortisol levels in biofluids from early Alzheimer's disease patients.

Cortisol dysregulation is proposed as a factor in the development of Alzheimer's disease (AD). AD patients can show high cortisol levels in prodromal phases of AD, early enough that neuropsychological alterations exist but activities of daily living remain unimpaired. Nevertheless, it is unknown if biofluid cortisol levels can have some AD predictive power together with neuropsychological assessment in prodromal stages in comparison with other cognitive disorders. In this work, an analytical method based on ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) was applied to determine the cortisol levels in different biofluids (urine, plasma, saliva, cerebrospinal fluid). Early AD patients and non-AD patients recruited at out-patient neurological unit were classified from the standard cerebrospinal fluid biomarkers levels (β-amyloid, tau, phosphorylated tau), and studied with an extensive neuropsychological assessment including global, neuropsychological, functional and affective scales. We used a logistic regression model to discriminate between the AD and non-AD groups. Higher plasma cortisol levels were found in the AD group than in the non-AD group (p < 0.001). Regarding neuropsychological evaluation, delayed memory was used as representative of the neuropsychological status, and lower scores were obtained in the AD group (p < 0.001). The prediction model, including plasma cortisol levels and delayed memory scores, achieved an AUC of 0.93, as well as a sensitivity of 97% and a specificity of 69.4%. In conclusion, plasma cortisol levels and delayed memory scores were specifically impaired in early AD, allowing the development of a new diagnostic model which could be employed as a very satisfactory screening system.