Is interleukin 6 measurement in exhaled breath of COVID-19 and post-COVID-19 lung fibrosis patients easy, sensitive and useful indicator of intensity of airway inflammation and fibrosis?

COVID-19 has emerged as a global pandemic. It mainly manifests as pneumonia, which may deteriorate into severe respiratory failure. The major hallmark of the disease is the systemic inflammatory immune response characterized by cytokine storm (CS). CS is marked by elevated levels of inflammatory cytokines, mainly interleukin-6 (IL-6), IL-8, IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFNγ). Of these, IL-6 is found to be significantly associated with higher mortality. IL-6 is also a robust marker for predicting disease prognosis and deterioration of the clinical profile. (1) IL-6 was detectable in the breath condensate of all healthy nonsmokers but was significantly higher in COPD patients. Exhaled breath condensate is totally noninvasive and highly acceptable to patients. The collection procedure has no effect on airway function or inflammation, and there is growing evidence that abnormalities in condensate composition may reflect biochemical changes in airway lining fluid. This method has been successfully used in previous studies to investigate several inflammatory markers in COPD and asthmatic patients. (2) Il-6 is produced in the lung by interstitial fibroblasts, alveolar macrophages, and large-vessel and bronchial epithelial cells.These studies and our novel method after clinical trials may open the field for future therapies for COVID-19 and post-COVID-19 lung fibrosis by inhaler transport medicines as a new challenge for overcoming sequelae of this pandemic. This suggested a new procedure for the measurement of exhaled IL-6. We studied which IL-6 is risky is IL-6 produced from airway or endocrine IL-6 or immune IL-6. We can classify IL-6 as cytokine storm under pathological conditions to three mains of sources as immune IL-6 produced from respiratory system like interstitial fibroblasts and bronchial epithelial cells and fibroblast, second endocrine IL-6 produced from zona glomerulosa of adrenal glands stimulated by ACTH and immune IL-6 from macrophages and other immune cells. From previous studies we need a procedure acceptable to patients and easy, noninvasive, sensitive so we plan for clinical trial registered at clinicaltrials.gov about measurement of interleukin-6 at exhaled condensate of covid-19 patients and post-19 covid patients with lung fibrosis as a novel study which may be a useful tool, easy, sensitive for early intervention with anti-il6 medications and determine the degree of severity by inflammatory markers for intensity of airway inflammation.

Cellular Senescence in Adrenocortical Biology and Its Disorders

Cellular senescence is considered a physiological process along with aging and has recently been reported to be involved in the pathogenesis of many age-related disorders. Cellular senescence was first found in human fibroblasts and gradually explored in many other organs, including endocrine organs. The adrenal cortex is essential for the maintenance of blood volume, carbohydrate metabolism, reaction to stress and the development of sexual characteristics. Recently, the adrenal cortex was reported to harbor some obvious age-dependent features. For instance, the circulating levels of aldosterone and adrenal androgen gradually descend, whereas those of cortisol increase with aging. The detailed mechanisms have remained unknown, but cellular senescence was considered to play an essential role in age-related changes of the adrenal cortex. Recent studies have demonstrated that the senescent phenotype of zona glomerulosa (ZG) acts in association with reduced aldosterone production in both physiological and pathological aldosterone-producing cells, whereas senescent cortical-producing cells seemed not to have a suppressed cortisol-producing ability. In addition, accumulated lipofuscin formation, telomere shortening and cellular atrophy in zona reticularis cells during aging may account for the age-dependent decline in adrenal androgen levels. In adrenocortical disorders, including both aldosterone-producing adenoma (APA) and cortisol-producing adenoma (CPA), different cellular subtypes of tumor cells presented divergent senescent phenotypes, whereby compact cells in both APA and CPA harbored more senescent phenotypes than clear cells. Autonomous cortisol production from CPA reinforced a local cellular senescence that was more severe than that in APA. Adrenocortical carcinoma (ACC) was also reported to harbor oncogene-induced senescence, which compensatorily follows carcinogenesis and tumor progress. Adrenocortical steroids can induce not only a local senescence but also a periphery senescence in many other tissues. Therefore, herein, we systemically review the recent advances related to cellular senescence in adrenocortical biology and its associated disorders.

Signal Transduction of Mineralocorticoid and Angiotensin II Receptors in the Central Control of Sodium Appetite: A Narrative Review

Sodium appetite is an innate behavior occurring in response to sodium depletion that induces homeostatic responses such as the secretion of the mineralocorticoid hormone aldosterone from the zona glomerulosa of the adrenal cortex and the stimulation of the peptide hormone angiotensin II (ANG II). The synergistic action of these hormones signals to the brain the sodium appetite that represents the increased palatability for salt intake. This narrative review summarizes the main data dealing with the role of mineralocorticoid and ANG II receptors in the central control of sodium appetite. Appropriate keywords and MeSH terms were identified and searched in PubMed. References to original articles and reviews were examined, selected, and discussed. Several brain areas control sodium appetite, including the nucleus of the solitary tract, which contains aldosterone-sensitive HSD2 neurons, and the organum vasculosum lamina terminalis (OVLT) that contains ANG II-sensitive neurons. Furthermore, sodium appetite is under the control of signaling proteins such as mitogen-activated protein kinase (MAPK) and inositol 1,4,5-thriphosphate (IP3). ANG II stimulates salt intake via MAPK, while combined ANG II and aldosterone action induce sodium intake via the IP3 signaling pathway. Finally, aldosterone and ANG II stimulate OVLT neurons and suppress oxytocin secretion inhibiting the neuronal activity of the paraventricular nucleus, thus disinhibiting the OVLT activity to aldosterone and ANG II stimulation.

Dichlorodiphenyltrichloroethane and the Adrenal Gland: From Toxicity to Endocrine Disruption

Endocrine disruptors are exogenous compounds that pollute the environment and have effects similar to hormones when inside the body. One of the most widespread endocrine disruptors in the wild is the pesticide dichlorodiphenyltrichloroethane (DDT). Toxic doses of DDT are known to cause cell atrophy and degeneration in the adrenal zona fasciculata and zona reticularis. Daily exposure in a developing organism to supposedly non-toxic doses of DDT have been found to impair the morphogenesis of both the cortex and the medulla of the adrenal glands, as well as disturbing the secretion of hormones in cortical and chromaffin cells. Comparison of high and very low levels of DDT exposure revealed drastic differences in the morphological and functional changes in the adrenal cortex. Moreover, the three adrenocortical zones have different levels of sensitivity to the disruptive actions of DDT. The zona glomerulosa and zona reticularis demonstrate sensitivity to both high and very low levels of DDT in prenatal and postnatal periods. In contrast, the zona fasciculata is less damaged by low (supposedly non-toxic) exposure to DDT and its metabolites but is affected by toxic levels of exposure; thus, DDT exerts both toxic and disruptive effects on the adrenal glands, and sensitivity to these two types of action varies in adrenocortical zones. Disruptive low-dose exposure leads to more severe affection of the adrenal function.

Biological Sex Modifies Aldosterone’s Secretion at a Cellular Level

Inconsistencies have been reported on the effect of sex on aldosterone levels leading to clinical confusion. The reasons for these inconsistencies, are uncertain but include: estrogen and/or its receptor modulating target gene responses to mineralocorticoid receptor activation and aldosterone secretagogues’ levels. This study’s goal was to determine whether aldosterone’s biosynthesis also differed by sex. Two approaches were used. First, plasma renin activity (PRA) and aldosterone were measured in rats. Both were significantly higher in males. Secondly, using rat zona glomerulosa (ZG) cells, we assessed three ex-vivo areas:1) activity/levels of early steps in aldosterone’s biosynthesis (StAR and CYP11A1); 2) activity/levels of a late step (CYP11B2); and 3) the status of the MR mediated, ultrashort-feedback-loop. Females had higher expression of CYP11A1 and StAR; and increased CYP11A1 activity (increased pregnenolone/ corticosterone levels) but did not differ in CYP11B2 expression or activity (aldosterone/ levels). Activating the ZG’s MR (thereby activating the ultrashort-feedback-loop) reduced CYP11B2’s activity similarly in both sexes. Ex-vivo, these molecular effects were accompanied, in females, by lower aldosterone basally but higher aldosterone with angiotensin II stimulation. In conclusion, we documented that not only was there a sex-mediated difference in the activity of aldosterone’s biosynthesis, but also these differences at the molecular level, help explain the variable reports on aldosterone’s circulating levels. Basally, both in-vivo and ex-vivo, males had higher aldosterone levels, likely secondary to higher aldosterone secretagogue levels. However, in response to acute stimulation, aldosterone levels are higher in females because of the greater levels and/or activity of their StAR/CYP11A1.

Colocalization of Wnt/β-catenin and ACTH signaling pathways and paracrine regulation in aldosterone producing adenoma

Context Aldosterone-producing adenomas (APA) are a common cause of primary aldosteronism. Despite the discovery of somatic mutations in APA and characterization of multiple factors regulating adrenal differentiation and function, the sequence of events leading to APA formation remains to be determined. Objective We investigated the role of Wnt/β-catenin and ACTH signaling, as well as elements of paracrine regulation of aldosterone biosynthesis in adrenals with APA and their relationship to intratumoral heterogeneity and mutational status. Design We analyzed expression of CYP11B2, CYP17A1, β-catenin, MC2R, pCREB, Tryptase, S100, CD34 by multiplex immunofluorescence and IHC guided RT-qPCR. Setting 11 adrenals with APA and one with micronodular hyperplasia from patients with PA were analysed. Main Outcome Measure(s) Localization of CYP11B2, CYP17A1, β-catenin, MC2R, pCREB, Tryptase, S100, CD34 in APA and aldosterone producing cell clusters (APCC). Results Immunofluorescence revealed abundant mast cells and a dense vascular network in APA, independent of mutational status. Within APA, mast cells were localized in areas expressing CYP11B2 and were rarely co-localized with nerve fibers, suggesting that their degranulation is not controlled by innervation. In these same areas, ß-catenin was activated, suggesting a zona glomerulosa cell identity. In heterogeneous APA with KCNJ5 mutations, MC2R and VEGFA expression was higher in areas expressing CYP11B2. A similar pattern was observed in APCC, with high expression of CYP11B2, activated β-catenin, and numerous mast cells. Conclusions Our results suggest that aldosterone producing structures in adrenals with APA share common molecular characteristics and cellular environment, despite different mutation status, suggesting common developmental mechanisms.

Adrenal glands of mice and rats: A comparative morphometric study

Mice and rats are among the most used laboratory animals. They share numerous similarities along with differences, some yet unexplored. One of them is the morphometry of their adrenal glands, whose characteristics may be related to differences in energy management, immune response, drug metabolism, behaviour and temperament. The present study tries to fill this knowledge gap with the evaluation and comparison of adrenal gland anatomical/morphometric parameters of mice and rats. In groups of 10 ( n = 10) adult, male and female BALB/c mice and Wistar rats, one in every 20 sections transverse to the longitudinal axis of the gland was used for measuring entire gland area, capsule, entire cortex, cortex zones and medulla with the aid of an image analysis system and subjected to statistical analysis. Quotients of the individual areas were calculated and comparison between the resulting ratios was performed. Gland length and volume were also calculated. Statistically significant differences were revealed between the rat female and male cortex area, rat and mouse medulla/cortex, medulla/gland, zona glomerulosa/cortex and cortex/gland ratios, male and female rats’ medulla/cortex, medulla/gland, capsule/gland, zona glomerulosa/cortex, zona reticularis/cortex and zona glomerulosa/zona fasciculata ratios, length and volume. The correlation evaluation revealed that in male rats and in female mice the larger medulla area was accompanied by a larger cortex area and vice versa. In general, a larger cortex area was accompanied by larger areas of cortex zones. The collected data and the revealed differences can possibly contribute to the understanding of the physiology of the two species.

HIPERALDOSTERONISMO EM FELINOS

Introdução: O hiperaldosteronismo é uma endocrinopatia onde ocorre o aumento da produção do hormônio aldosterona, secretado pela zona glomerulosa da glândula adrenal. Este hormônio tem como função regular da homeostase de eletrólitos, principalmente o sódio e potássio, fazendo a manutenção da pressão arterial. Objetivos: Apresentar principais alterações sintomatológicas causadas pelo hiperaldosteronismo em felinos, assim como seu diagnóstico e tratamento, através de uma revisão bibliográfica, a fim de salientar a importância deste diagnóstico diferencial. Material e métodos: Foram analisados estudos e dados publicados sobre a enfermidade, relatados em artigos científicos e trabalhos de conclusão de curso. Resultados: O hiperaldosteronismo primário ocorre devido alterações produzidas por neoplasias e hiperplasias na glândula adrenal, no qual causam a maior secreção de aldosterona, levando assim a uma maior absorção de sódio e excreção de potássio, causando um aumento na volemia e alterações bioquímicas, como hipocalemia, hipomagnesemia, hipofosfatemia, hipocloremia e aumento de ureia e creatinina. As principais alterações perceptíveis em felinos, são a hipertensão sistêmica, poliomiopatia hipocalêmica, ventroflexão cervical, mialgia, anorexia, alterações oculares, entre outros. Já o hiperaldosteronismo secundário, pode ser causado por alterações como desidratação, hipotensão, deficiência de sódio, entre outros, sendo também associados a insuficiência renal, hepática e cardíaca, na qual gera a ativação do sistema renina-angiotensina-aldosterona, sendo estes encontrados com valores alterados. Para o diagnóstico, é recomendado a dosagem de aldosterona, potássio e renina plasmática, auxiliando assim na distinção entre o hiperaldosteronismo primário e secundário, indicando assim, se a renina plasmática estiver diminuída, que os valores elevados de aldosterona são pouco estimulados pelo sistema renina-angiotensina. É necessário também outros exames complementares, como a ultrassonografia, tomografia ou ressonância magnética, para constatar a presença de massas nas adrenais. O tratamento clínico de ambas as formas, consiste no tratamento sintomático e o tratamento cirúrgico é indicado quando há tumores na adrenal, sendo recomendado a adrenalectomia unilateral. Conclusão: O hiperaldosteronismo é uma doença em que se manifesta através da hipocalemia e hipertensão, podendo causar severas alterações em felinos. Sendo assim é importante estar atento aos sintomas para que possa ser feito o tratamento correto e eficaz ao paciente.

Latent Adrenal Insufficiency: From Concept to Diagnosis

Primary adrenal insufficiency (PAI) is a rare disease and potentially fatal if unrecognized. It is characterized by destruction of the adrenal cortex, most frequently of autoimmune origin, resulting in glucocorticoid, mineralocorticoid, and adrenal androgen deficiencies. Initial signs and symptoms can be nonspecific, contributing to late diagnosis. Loss of zona glomerulosa function may precede zona fasciculata and reticularis deficiencies. Patients present with hallmark manifestations including fatigue, weight loss, abdominal pain, melanoderma, hypotension, salt craving, hyponatremia, hyperkalemia, or acute adrenal crisis. Diagnosis is established by unequivocally low morning serum cortisol/aldosterone and elevated ACTH and renin concentrations. A standard dose (250 µg) Cosyntropin stimulation test may be needed to confirm adrenal insufficiency (AI) in partial deficiencies. Glucocorticoid and mineralocorticoid substitution is the hallmark of treatment, alongside patient education regarding dose adjustments in periods of stress and prevention of acute adrenal crisis. Recent studies identified partial residual adrenocortical function in patients with AI and rare cases have recuperated normal hormonal function. Modulating therapies using rituximab or ACTH injections are in early stages of investigation hoping it could maintain glucocorticoid residual function and delay complete destruction of adrenal cortex.

The Herbicide Atrazine Potentiates Angiotensin II-Induced Aldosterone Synthesis and Release From Adrenal Cells

Atrazine is one of the most commonly used pre-emergence and early post-emergence herbicides in the world. We have shown previously that atrazine does not directly stimulate the pituitary or adrenal to trigger hormone release but acts centrally to activate a stress-like activation of the hypothalamic-pituitary-adrenal axis. In doing so, atrazine treatment has been shown to cause adrenal morphology changes characteristic of repeated stress. In this study, adrenals from atrazine treated and stressed animals were directly compared after 4 days of atrazine treatment or restraint stress. Both atrazine and stressed animals displayed reduced adrenocortical zona glomerulosa thickness and aldosterone synthase (CYP11B2) expression, indicative of repeated adrenal stimulation by adrenocorticotropic hormone. To determine if reduced CYP11B2 expression resulted in attenuated aldosterone synthesis, stressed and atrazine treated animals were challenged with angiotensin II (Ang II). As predicted, stressed animals produced less aldosterone compared to control animals when stimulated. However, atrazine treated animals had higher circulating aldosterone concentrations compared to both stressed and control groups. Ang II-induced aldosterone release was also potentiated in atrazine pretreated human adrenocortical carcinoma cells (H295R). Atrazine pretreated did not alter the expression of the rate limiting steroidogenic StAR protein or angiotensin II receptor 1. Atrazine treated animals also presented with higher basal blood pressure than vehicle treated control animals suggesting sustained elevations in circulating aldosterone levels. Our results demonstrate that treatment with the widely used herbicide, atrazine, directly increases stimulated production of aldosterone in adrenocortical cells independent of expression changes to rate limiting steroidogenic enzymes.