Maternal and neonatal canine cortisol measurement in multiple matrices during the perinatal period: A pilot study

Stress exposure during perinatal period may lead to maternal cortisol increase that negatively affects the offspring development. In recent years, the interest on non-invasive sampling methods to measure cortisol as a marker of stress is increasing in both humans and animals. Indeed, discomfort due to blood collection may compromise the diagnostic outcome, mainly in uncooperative patients. So far, some alternative matrices but not milk have been explored in adult dogs, while no data are available on the neonate and paediatric live pups. This study aimed to measure cortisol concentration in different biological substrates in both dams (blood, saliva, hair and milk) and pups (saliva and hair) at established times from proestrus up to two months after parturition. For this purpose, five female German shepherd bitches and their 22 pups were enrolled. Cortisol concentration was assessed using the enzyme immunoassay kit (Salivary Cortisol ELISA kit, Salimetrics) after matrices appropriate preparation if required. Cortisol was measurable in all the substrates, except some milk samples below the detection limit. Maternal cortisol concentrations differed among the matrices (P <0.0001) with the highest values recorded in plasma (median 0.596 μg/dL) compared to saliva (median 0.159 μg/dL), hair (median 0.083 μg/dL) and milk (median 0.045 μg/dL). Cortisol in dams did not vary within the same matrix over time. In pups, salivary (median 0.295 μg/dL) cortisol was always higher than hair (median 0.049 μg/dL; P <0.0001). At birth (P = 0.01) and two months later (P = 0.05), neonatal salivary cortisol was higher compared to other samplings. The present study demonstrates the suitability of these innovative substrates for cortisol measurement, suggesting them as potential diagnostic support in canine neonatology and welfare.

Desmopressin Test in Silent Corticothoph Pituitary Adenomas, a Preliminary Study - a Comparison With Cushing Disease

Introduction: Silent corticothop adenomas (SCAs) account for 9 to 19% of nonfunctioning pituitary adenomas and behave as the most agressive pituitary tumors with more invasiveness and high recurrence rate. The identification of these patients during the preoperative stage could predict better surgery results. Some authors refer to high basal ACTH level in the preoperative evaluation as the only marker but until this date, there are no clinical and hormonal markers that could predict its occurrence. The aim of this study is to evaluate the response to desmopressin test and the presence of silent corticothoph tumors. Patients and Methods: Among 496 patients underwent pituitary surgery, 425 were pituitary adenomas, (106 were acromegaly, 46 Cushing disease, 10 prolactinomas, 263 nonfunctioning adenomas). Twenty-three patients, 17 with Cushing disease, (CD), mean age: 31 y, 13 female and 4 male) and 6 patients with SCA (mean age: 47 y, 5 female and 1 male) had positive ACTH confirmed by immunohistochemical analysis. Clinical characteristics: in the CD group, 53% had hypertension (9/17), 42% diabetes (7/17), 100% dyslipidemia, BMI was 30.7 kg/m2. Among SCA group, 67% hypertension, 50% diabetes, 50% dyslipidemia, BMI was 28 kg/m2. All patients were evaluated with basal ACTH and DHEAS before surgery. Patients with SCA underwent desmopressin test and were compared to CD. Dexamethasone suppression test (DST 1 mg) and 24-hour free urinary test was performed in patients with CD and in two patients with SCA. Response to desmopressin test was considered positive when increase in cortisol was above 20% and in ACTH of 35% using chemiluminescence assay (Immulite 2000). Results: Among CD group, the median (med) basal ACTH was 75.9 pg/mL (30.9 to 211), the med basal cortisol was 22.5 µg/dL (14.5 to 43.5), the med DHEAS was 170 µg/dL (33 to 465), the med 24h urinary free cortisol of 454.5 µg/24 h (149 to 1673) and med basal cortisol after DST 1mg of 15.4 µg/dL (4.7 to 31.5). Among SCA, med basal ACTH was 19.4 pg/mL (9.5 to 65.5), the med basal cortisol was 9.5 µg/dL (7.8 to 16.4) and the med DHEAS was 104.5 µg/dL (82 to 127). Only 4 patients with CD had macroadenomas. All of them responded with ACTH increase (med increase of 98%, 31.6 to 377%), and only 4 did not respond to cortisol increase (med increase of 54.4%, 0 to 167%). All patients with SCA had macroadenomas. Only one patient did not respond to ACTH increase (med increase of 123.5%, 9.5 to 1522%, 9.5 to 1522%), and 3 patients did not respond to cortisol increase (med increase of 17.9%, 0 to 234%). Discussion: SCA are invasive tumors, with high recurrence and tests predicting their occurrence are missing. We hypothesized that as ACTH is present in the adenoma a response to desmopressin test could exist (like CRH). Conclusion:The desmopressin test can be a useful tool in the evaluation of SCA and can predict pathological phenotype in preoperative tumors.

Association of stress-related limbic activity and baseline interleukin-6 plasma levels in healthy adults

There is a bidirectional interaction between peripheral inflammatory processes and the brain. In the context of stress, top-down effects on the immune system have already been studied, but bottom-up effects of peripheral inflammation on neural processes are still unclear. In the present study, baseline plasma levels of the cytokine interleukin (IL)-6 were obtained as a measure of peripheral inflammatory processes and psychosocial stress was induced in 78 healthy adults during functional magnetic resonance imaging. The association of IL-6 with stress-related neural activation in the amygdala, anterior insula, and anterior cingulate cortex as regions of interest was explored. In order to account for individual differences in the stress response, salivary cortisol increase was also considered in the analyses. Stress task-related neural activity in the amygdala and anterior insula was significantly and negatively associated with IL-6 levels only in participants showing a cortisol response. These results suggest that peripheral inflammatory processes might affect the evaluation of a social stressor and highlight that individual differences in stress task responses should be considered in future studies.

The Danger Within: Covid-19 Affinity for ACE2 receptors in Adipose Tissue and Testes. The Protective Effects of Estradiol, Fitness and Weight Management

The imminent danger of the Covid-19 pandemic has accelerated research in pharmaceuticals designed to interfere with the virus’ entry into the body via ACE2 receptors, or the viral RNA replication that often overwhelms immune defences. The scope of this review was to elucidate the main human vulnerabilities like certain organs’ enrichment in ACE2 receptors increasing viral affinity to males, the aged and certain pre-existing conditions including diabetes, CVD and pulmonary diseases, that deteriorate with increasing obesity, inflammation and toxicity. The current perspective focuses on the primary components of dysregulated health predisposing individuals to Covid-19, including hormonal imbalance,increased lipids and lipoproteins, thyroid dysfunction, degraded fitness, and age-related testosterone decline accompanied by cortisol increase that provokes stress eating behaviours and weight accumulation.We examined the molecular dynamics illustrating the action of new therapeutics necessary for Covid-19 patients; theestradiol advantage hypothesis; alternative therapies including hormone replacement procedures andmesenchymal stem cells; plus preventive and protective interventions. Obesity increases the probability of Covid-19 infection due to its abundance of ACE2 receptors.Physical activitymay decrease Covid-19 vulnerability, due to the diminished ACE-2 expression in the muscle. There are a number the fat managementsolutions featuring lasers and radiofrequency which, however do not enhance fitness. Seven recently published clinical studies with a total of 95 subjects, 73 males and 22 females, demonstrated visceral fat reduction combined with increasedskeletal muscle mass. A metanalysis performed on their data revealeda statistically significant decrease in several variables including BMI, lipids, lipoproteins, toxicityand inflammation as measured by CRP, Creatinine and Bilirubin, andoptimally healthier levels ofCortisol, Testosterone, Free T3,IGF-1, Insulin, and the appetite controlling hormones Leptin and Ghrelin.

Stress-Induced Alteration in Chloride Transporters in the Trigeminal Nerve May Explain the Comorbidity between Depression and Migraine

Migraine is frequently comorbid with depression and anxiety disorders. In the case of depression and panic disorder, the associations seem to be bidirectional. Stress (activation of the hypothalamic-pituitary-adrenal axis) is thought to be involved in increasing the attack frequency. In the current review, it is argued that elevated levels of cortisol increase the function of chloride-ion transporter NKCC1 and decrease the function of chloride-extruder KCC2 in the trigeminal nerve. This leads to a diminished inhibitory effect of gamma-aminobutyric acid (GABA) and an enhanced likelihood of a migraine attack. Since migraine attacks themselves are stressful, and since brain areas are activated that could contribute to panic, anxiety and depression, a number of self-sustaining circular processes could occur that would explain the bi-directionality of the associations. On the basis of this hypothesis, several novel therapeutic approaches to counter the pathological process can be proposed. These include inhibition of corticotrophin releasing factor by CRF1 receptor antagonists, blockade of adrenocorticotropic hormone (ACTH) at the MC2 receptor, and inhibition of the hyperactive NKCC1 chloride-transporter.

Non-invasive Diagnostic Strategy in ACTH-dependent Cushing’s Syndrome

Context Inferior petrosal sinus sampling (IPSS) is used to diagnose Cushing’s disease (CD) when dexamethasone-suppression and CRH tests, and pituitary magnetic resonance imaging (MRI), are negative or give discordant results. However, IPSS is an invasive procedure and its availability is limited. Objective To test a noninvasive diagnostic strategy associated with 100% positive predictive value (PPV) for CD. Design Retrospective study. Setting Two university hospitals. Patients A total of 167 patients with CD and 27 patients with ectopic ACTH-syndrome investigated between 2001 and 2016. Main Outcome Measure(s) Performance of a strategy involving the CRH and desmopressin tests with pituitary MRI followed by thin-slice whole-body computed tomography (CT) scan in patients with inconclusive results. Results Using thresholds of a cortisol increase &gt; 17% with an ACTH increase &gt; 37% during the CRH test and a cortisol increase &gt; 18% with an ACTH increase &gt; 33% during the desmopressin test, the combination of both tests gave 73% sensitivity and 98% PPV of CD. The sensitivity and PPV for pituitary MRI were 71% and 99%, respectively. CT scan identified 67% EAS at presentation with no false-positives. The PPV for CD was 100% in patients with positive responses to both tests, with negative pituitary MRI and CT scan. The Negative Predictive Value was 100% in patients with negative responses to both tests, with negative pituitary MRI and positive CT scan. Using this strategy, IPPS could have been avoided in 47% of patients in whom it is currently recommended. Conclusions In conjunction with expert radiologic interpretation, the non-invasive algorithm studied significantly reduces the need for IPSS in the investigation of ACTH-dependent Cushing’s syndrome.

Stress affects the prediction of others’ behavior

Predicting behavior of other people is vital for successful social interactions. We tested whether a stress-induced cortisol increase affects healthy young men’s prediction of another individual’s behavior. Forty-two participants were randomly assigned to a stress or a control condition. Afterwards they participated in a modified false-belief task that not only tests false-belief understanding but also the tendency to predict another person’s future behavior based on his former behavior. Subjective ratings and salivary cortisol concentrations revealed a successful stress induction. Stress did not affect participants’ attribution of false beliefs but it increased the probability to predict that a protagonist would act according to his former behavior. Recognizing that stress fosters the interpretation of others’ behavior following their former behavior and not their current goals extends previous research showing that stress fosters our own habitual behavior.

MON-295 Desmopressin Test in Silent Corticothoph Pituitary Adenomas, a Preliminary Study - a Comparison with Cushing Disease

Introduction: Silent corticothop adenomas (SCAs) account for 9 to 19% of nonfunctioning pituitary adenomas and behave as the most agressive pituitary tumors with more invasiveness and high recurrence rate. The identification of these patients during the preoperative stage could predict better surgery results. Some authors refer to high basal ACTH level in the preoperative evaluation as the only marker but until this date, there are no clinical and hormonal markers that could predict its occurrence. The aim of this study is to evaluate the response to desmopressin test and the presence of silent corticothoph tumors. Patients and methods: Among 475 patients with pituitary lesion, which underwent surgery, 332 were pituitary adenomas, (82 were acromegaly, 40 Cushing disease, 178 nonfunctioning adenomas). Twenty-three patients, 17 with Cushing disease, (CD), mean age: 31 y, 13 female and 4 male) and 6 patients with SCA (mean age: 47 y, 5 female and 1 male) had positive ACTH confirmed by immunohistochemical analysis. Clinical characteristics: in the CD group, 53% had hypertension (9/17), 42% diabetes (7/17), 100% dyslipidemia, BMI was 30.7 kg/m2. Among SCA group, 67% hypertension, 50% diabetes, 50% dyslipidemia, BMI was 28 kg/m2. All patients were evaluated with basal ACTH and DHEAS before surgery. Patients with SCA underwent desmopressin test and were compared to CD. Dexamethasone suppression test (DST 1 mg) and 24-hour free urinary test was performed in patients with CD and in two patients with SCA. Response to desmopressin test was considered positive when increase in cortisol was above 20% and in ACTH of 35% using chemiluminescence assay (Immulite 2000). Results: Among CD group, the median (med) basal ACTH was 75.9 pg/mL (30.9 to 211), the med basal cortisol was 22.5 µg/dL (14.5 to 43.5), the med DHEAS was 170 µg/dL (33 to 465), the med 24h urinary free cortisol of 454.5 µg/24 h (149 to 1673) and med basal cortisol after DST 1mg of 15.4 µg/dL (4.7 to 31.5). Among SCA, med basal ACTH was 19.4 pg/mL (9.5 to 65.5), the med basal cortisol was 9.5 µg/dL (7.8 to 16.4) and the med DHEAS was 104.5 µg/dL (82 to 127). Only 4 patients with CD had macroadenomas. All of them responded with ACTH increase (med increase of 98%, 31.6 to 377%), and only 4 did not respond to cortisol increase (med increase of 54.4%, 0 to 167%). All patients with SCA had macroadenomas. Only one patient did not respond to ACTH increase (med increase of 123.5%, 9.5 to 1522%, 9.5 to 1522%), and 3 patients did not respond to cortisol increase (med increase of 17.9%, 0 to 234%). Discussion: SCA are invasive tumors, with high recurrence and tests predicting their occurrence are missing. We hypothesized that as ACTH is present in the adenoma a response to desmopressin test could exist (like CRH). Conclusion: The desmopressin test can be a useful tool in the evaluation of SCA and can predict pathological phenotype in preoperative tumors.