Epinephrine Release Research Articles

Impact of Chronic Moderate Psychological Stress on Skin Aging: Exploratory Clinical Study and Cellular Functioning.

Skin is continuously exposed to environmental external and internal factors, including psychological stress (PS). PS has been reported to trigger different dermatoses such as psoriasis, atopic dermatitis, vitiligo, alopecia areata, and acne through the release of cortisol and epinephrine. To clinically explore PS-induced measurable skin aging signs in subjects with moderate versus mild chronic PS, and to investigate the effect of chronic PS on DNA damage at cellular level. In vitro stress tests with cortisol and epinephrine, and with cortisol only on extracellular matrix (ECM) synthesis, as well as on normal human skin fibroblast and keratinocyte functioning, including skin barrier and wound healing were performed. Moderately stressed subjects in the context of the clinical study had a significantly decreased antioxidant potential and impacted skin barrier integrity, as well as significantly increased signs of microrelief alterations (skin texture and fine lines) reaching an increased severity of about 32.9%. At a cellular level, DNA integrity, ECM synthesis, wound healing, and skin barrier parameters were impacted by increased stress hormone levels. The clinical exploratory studies presented herewith, as well as the study of cell functioning under stress, have provided evidence that chronic PS significantly affects skin homeostasis and triggers skin aging.

Sublingual route for the continuous release of epinephrine from chitosan nanoparticles

Administration of Epinephrine (Epi) using an intramuscular autoinjector (EAI) is the primary first aid treatment available for the management of anaphylaxis in the community of many countries. Around 1 in 10 patients require a second EAI within 5–15 min. Previously, we successfully developed rapidly disintegrating sublingual tablets (RDST) containing Epi microcrystals that exhibited comparable in vivo pharmacokinetics to that of EAI to overcome the drawbacks of EAIs. To further advance these RDST, we hypothesized that synthesizing Epi nanoparticles (Epi-NP) for incorporation into our previously developed Epi RDST could prolong Epi release sufficiently to eliminate the need for a second dose. Therefore, the primary objective of this research was to synthesize various Epi-NP with different drug loads and investigate their release profiles and sublingual permeability. In vitro release and ex vivo permeability studies were performed using synthetic dialysis membranes and excised porcine sublingual membranes, receptively. The release and permeation of Epi from both NP with adsorbed Epi (Ads Epi-NP) and NP with encapsulated Epi (Enc Epi-NP) were significantly slowed down and achieved continuous control over 8 h. At 60 % Epi load, Epi release from Enc Epi-NP was significantly lower than from Ads Epi-NP, however, the cumulative permeated Epi was only significantly lower towards the end of the experiment, suggesting similar control over Epi permeability. Finally, visualization of NP permeation across the sublayers of the sublingual mucosa confirmed the formation of a temporary depot for Epi and maintained Epi release for several hours post-administration. The results from this study demonstrate for the first time the short-term sustained-release potential of the sublingual route to overcome the need for a second dose.

Chronic stimulation desensitizes β2-adrenergic receptor responses in natural killer cells.

Adrenergic receptors (ARs) are preferentially expressed by innate lymphocytes such as natural killer (NK) cells. Here, we study the effect of epinephrine-mediated stimulation of the β2-adrenergic receptor (β2AR) on the function of human NK cells. Epinephrine stimulation inhibited early NK cell signaling events and blocked the function of the integrin LFA-1. This reduced the adhesion of NK cells to ICAM-1, explaining how NK cells are mobilized into the peripheral blood upon epinephrine release during acute stress or exercise. Additionally, epinephrine stimulation transiently reduced NK cell degranulation, serial killing, and cytokine production and affected metabolic changes upon NK cell activation via the cAMP-protein kinase A (PKA) pathway. Repeated exposure to β2AR agonists resulted in the desensitization of the β2AR via a PKA feedback loop-initiated G-protein switch. Therefore, acute epinephrine stimulation of chronically β2AR stimulated NK cells no longer resulted in inhibited signaling and reduced LFA-1 activity. Sustained stimulation by long-acting β2-agonists (LABA) not only inhibited NK cell functions but also resulted in desensitization of the β2AR. However, peripheral NK cells from LABA-treated asthma patients still reacted unchanged to epinephrine stimulation, demonstrating that local LABA administration does not result in detectable systemic effects on NK cells.

AROMA THERAPY INTERVENTION TO IMPROVING SLEEP QUALITY IN PATIENTS WITH CARDIOVASCULAR PROBLEMS

Background: Poor sleep quality will lead to stressful conditions which can cause the release of epinephrine and norepinephrine has the effect of increasing the oxygen demand for the myocardium. Aromatherapy is recommended as a no-side-effect, non-invasive, inexpensive and painless method to reduce anxiety and pain. improve sleep quality of cardiovascular patients. Purpose: The aim of this study is to analyze the potential of aromatherapy interventions in patients with cardiovascular disorders. Methods: The research method in this article uses a literature study approach. The articles will be selected based on the following criteria. The search sources in this article use scientific-based databases (ScienceDirect, PubMed, EBSCOhost, and Wiley Library Online). The search starts from December 2022 using the keywords Cardiac Patients – Aromatherapy – Sleep quality. Result: Inhalation aromatherapy with peppermint or lavender essential oil can improve the sleep quality of cardiovascular patients. Inhaling aromatic oil stimulates the sense of smell affecting the central nervous system. Lavender oil can reduce fatigue, anxiety, heart rate, respiratory rate, and blood pressure, improve lung and brain oxygenation, and improve sleep quality. The use of essential oils can be a safe and cost-effective therapy for sleep disorders and can reduce the overuse of prescription medications. Implication: Lavender and peppermint essential oils have a therapeutic effect that relaxes the body. These effects can make the body feel relaxed and release hormones that can help patients sleep. However, this intervention still needs further research specifically from the aspect of dose and medium of aromatherapy.

Chromogranin A (CgA) Deficiency Attenuates Tauopathy by Altering Epinephrine-Alpha-Adrenergic Receptor Signaling.

Our previous studies have indicated that insulin resistance, hyperglycemia, and hypertension in aged wild-type (WT) mice can be reversed in mice lacking chromogranin-A (CgA-KO mice). These health conditions are associated with a higher risk of Alzheimer's disease (AD). CgA, a neuroendocrine secretory protein has been detected in protein aggregates in the brains of AD patients. Here, we determined the role of CgA in tauopathies, including AD (secondary tauopathy) and corticobasal degeneration (CBD, primary tauopathy). We found elevated levels of CgA in both AD and CBD brains, which were positively correlated with increased phosphorylated tau in the frontal cortex. Furthermore, CgA ablation in a human P301S tau (hTau) transgenic mice (CgA-KO/hTau) exhibited reduced tau aggregation, resistance to tau spreading, and an extended lifespan, coupled with improved cognitive function. Transcriptomic analysis of mice cortices highlighted altered levels of alpha-adrenergic receptors (Adra) in hTau mice compared to WT mice, akin to AD patients. Since CgA regulates the release of the Adra ligands epinephrine (EPI) and norepinephrine (NE), we determined their levels and found elevated EPI levels in the cortices of hTau mice, AD and CBD patients. CgA-KO/hTau mice exhibited reversal of EPI levels in the cortex and the expression of several affected genes, including Adra1 and 2, nearly returning them to WT levels. Treatment of hippocampal slice cultures with EPI or an Adra1 agonist intensified, while an Adra1 antagonist inhibited, tau hyperphosphorylation and aggregation. These findings reveal a critical role of CgA in regulation of tau pathogenesis via the EPI-Adra signaling axis.

Studying the role of cortisol in the pathogenesis of atopic dermatitis during pregnancy

Introduction. Atopic dermatitis (AD) is one of the most common dermatitis, characterized by complex pathogenetic mechanisms. Psychological stress is recognized as one of the triggers of AD. Stress causes a high release of cortisol and epinephrine or norepinephrine, stimulating the immune system, primarily T helper cells type 1 (Th1 cells), to produce pro-inflammatory cytokines, leading to a cellular immune response and inflammation. In recent years, there has been an increase in incidence among pregnant women, however, the specific mechanisms of the development of AD during pregnancy still remain poorly understood. Aim. To study the role of cortisol in AD during pregnancy.Materials and methods. The study included 76 pregnant women during an exacerbation of AD, 20 non-pregnant women during an exacerbation of AD, 20 non-pregnant women without AD, 20 pregnant women without AD. The severity of AD was determined using the SCORAD index. Cortisol levels were determined in blood serum using enzyme-linked immunosorbent assay (ELISA). Anxiety level was determined using the Beck Anxiety Inventory. The level of itching was determined using a 5D itching scale.Results. Cortisol levels in pregnant women with AD (629.8 pg/ml) were significantly higher than in non-pregnant women with AD (386.15 pg/ml) (p < 0.05). Cortisol levels were correlated with the severity level (Spearman coefficient = 0.203, p = 0.018), anxiety level (Spearman coefficient = 0.411, p = 0.001), and level of itching (Spearman coefficient = 0.352, p = 0.001).Conclusions. Cortisol is important in the pathogenesis of AD during pregnancy. During pregnancy with exacerbation of AD, higher values were observed than outside pregnancy.

Hypoglycemia Unawareness-A Review on Pathophysiology and Clinical Implications.

Hypoglycemia is a particular problem in people with diabetes while it can also occur in other clinical circumstances. Hypoglycemia unawareness describes a condition in which autonomic and neuroglycopenic symptoms of hypoglycemia decrease and hence are hardly perceivable. A failure to recognize hypoglycemia in time can lead to unconsciousness, seizure, and even death. The risk factors include intensive glycemic control, prior episodes of severe hypoglycemia, long duration of diabetes, alcohol consumption, exercise, renal failure, and sepsis. The pathophysiological mechanisms are manifold, but mainly concern altered brain glucose sensing, cerebral adaptations, and an impaired hormonal counterregulation with an attenuated release of glucagon, epinephrine, growth hormone, and other hormones, as well as impaired autonomous and neuroglycopenic symptoms. Physiologically, this counterregulatory response causes blood glucose levels to rise. The impaired hormonal counterregulatory response to recurrent hypoglycemia can lead to a vicious cycle of frequent and poorly recognized hypoglycemic episodes. There is a shift in glycemic threshold to trigger hormonal counterregulation, resulting in hypoglycemia-associated autonomic failure and leading to the clinical syndrome of hypoglycemia unawareness. This clinical syndrome represents a particularly great challenge in diabetes treatment and, thus, prevention of hypoglycemia is crucial in diabetes management. This mini-review provides an overview of hypoglycemia and the associated severe complication of impaired hypoglycemia awareness and its symptoms, pathophysiology, risk factors, consequences, as well as therapeutic strategies.

A Bilayer Microneedle for Modulated Sequential Release of Adrenaline and Lidocaine for Prolonged Local Anesthesia.

Chronic pain emerges as a major global health issue, significantly impacting individuals' health and quality of life. In this study, we designed a bilayer microneedle loaded with lidocaine nanocomposites in the inner layer and adrenaline (Adr) in the outer layer (HCP MNs) for modulated sequential release to achieve prolonged local anesthesia. The obtained HCP MNs featured an intact structure with adequate mechanical strength for efficient skin penetration. The bilayer structure of MNs was evidenced by loading two fluorescent dyes in each layer. Furthermore, these HCP MNs were capable of inducing rapid as well as prolonged local anesthetic effects in guinea pigs. Hence, the bilayer MN coloaded with Adr and lidocaine nanocomposite serves as a promising transdermal delivery platform for chronic pain management.

Mechanism of Caffeine and Challenges of Caffeine Dependence

By binding to adenosine A1 and A2A receptors, caffeine inhibits adenosine's inhibitory effect on neurons, thereby increasing alertness and improving concentration. At the same time, caffeine also stimulates the release of adrenaline and norepinephrine, further enhancing the sense of wakefulness. Caffeine also induces the release of dopamine and glutamate in the nucleus accumbens. They regulate human emotions, reward mechanisms, and memory. However, caffeine dependence can cause side effects such as insomnia, anxiety, heart palpitations, digestive problems, headaches, fatigue, and difficulty concentrating. To reduce dependency on caffeine, individuals can gradually decrease their intake, opt for decaffeinated tea or coffee, and ensure they have sufficient and restful sleep by adjusting sleep patterns. Maintaining overall health, a balanced diet, moderate exercise and effective coping with stress can help increase natural energy levels and reduce the need for caffeine. Seeking the help and guidance of a professional medical or mental health professional is also recommended for reducing caffeine dependence.

Identification of Subjective Fatigue accompanied by Hypertension in Female Workers Making Tiles in Indonesia: A Cross-Sectional Survey

Introduction: The home industry has production process activities that are carried out manually. The release of the hormone adrenaline increases blood pressure through an increase in heart rate and arterial contractions, thereby accelerating the feeling of tiredness. As a result, female workers risk getting tired faster than men. Fatigue can lead to a decrease in productivity, poor quality of work, negatively affect the safety of workers, and increase the risk of work accidents. It aims to analyze the risk factors for subjective fatigue in female workers who have hypertension. Methods: This type of quantitative research with a cross-sectional design. Respondents were 278 female hypertension workers at the Karangasem Village Tile Making Home Industry Center, Wirosari District, Grobogan, Indonesia. Data analysis using Chi-square with the SPSS 21. Results: Of a total of 278 female workers, most were married (85.3%), did not smoke (98.9%), did not consume alcohol (96%), worked longer than 8 hours (93.5%), worked more than five years (80.9%), heavy workload (80.6%). Subjective fatigue factors related such as alcohol consumption habits (0.019), exercise habits (0.000), body mass index (0.000), working period (0.000), and workload (0.000). Meanwhile, other risk factors were not related, age (0.315), marital status (0.744), education level (0.811), smoking habits (0.236), disease history (0.762), length of work (0.530), type of work (0.393) and work stress (0.451). Conclusion: Risk factors that have a relationship with subjective fatigue are workload, BMI, alcohol consumption habits, exercise habits, and length of service

SAT439 Euglycemic Diabetic Ketoacidosis When Combining Empagliflozin With Cocaine

Abstract Disclosure: F.A. Squicimari-de Cardenas: None. J. Bosques-Lorenzo: None. L. Alejandro: None. C. Rodriguez: None. C. Carlos: None. J.M. Colon Castellano: None. Background: Euglycemic DKA (eu-DKA) is a life-threatening emergency that can occur in patients with either type 1 or type 2 DM, and manifests as a milder degree of hyperglycemia with glucose levels around 200mg/dL. Cocaine abuse alone has been associated with DKA and eu-DKA by inducing cortisol, epinephrine, and norepinephrine release from adrenal glands, and by its anorexigenic effects by suppressing the feeding centers in the central nervous system. Here we present a case of eu-DKA in a patient that combined SGLT2i with cocaine. Clinical Case: A 66 y/o male with past medical history significant for DM type 2 treated with empagliflozin 25mg daily, metformin 1000mg BID, glargine 45 units am and pm, and exenatide 2mg. Patient presented to the ER with general malaise, dyspnea, abdominal pain, and nauseas. On initial evaluation, vital signs were remarkable for temperature at 97.8 F, pulse at 101 bpm, blood pressure 172/113 mmHg, respiratory rate at 20 bpm. Physical examination significant for a tachypneic, restless, and acutely ill patient that was alert, oriented and able to provide medical history. He and his family members denied him using illicit drugs or alcohol. Laboratories were found with severe metabolic acidosis. Central CO2 at 3.8 mEq/L (24-32 mEq/L), central glucose 241 mg/dL (70-99 mg/dL), elevated anion gap at 29.2 mEq/L, B-hydroxybutyrate markedly elevated at >83.3 mg/dL (0.2 - 2.81 mg/dL), arterial blood gases with pH at 7.11 (7.38-7.46) and HCO3 at 3.0 mmol/L (21-29 mmol/L). Urinalysis was positive for ketones and glucosuria. Toxicology came back positive for cocaine, and negative for alcohol. Due to patients’ severe acidosis and concerns for cardiac arrest, he was intubated for airway protection and admitted to the ICU for further management. DKA protocol was initiated and treatment with IV insulin, IV 5% Dextrose, 0.45% normal saline, potassium replacement, and IV bicarbonate provided until anion gap closed and acidosis resolved. Eventually, patient was successfully extubated and transferred to ward for further management and diabetic treatment optimization. Conclusion: Cocaine use is an independent trigger for DKA and eu-DKA. Euglycemic DKA has been raising with the surge of SGLT2i. As in our patient, concomitant use of SGLT2i and cocaine can precipitate patients for development for DKA/eu-DKA. It is important to educate patients to avoid concomitant use of this two and to perform toxicology screening in patients presenting with DKA or eu-DKA. Presentation: Saturday, June 17, 2023

Bioinspired CuZn-N/C Single-Atom Nanozyme with High Substrate Specificity for Selective Online Monitoring of Epinephrine in Living Brain.

Though many elegant laccase mimics have emerged, these mimics generally have no substrate selectivity as well as low activity, making it difficult to fulfill the demand for monitoring in physiological conditions. Herein, inspired by the Cu-N ligand structure in the active site of natural laccase, we revealed that a carbon nanomaterial with atomically dispersed Cu and Zn atoms (CuZn-N/C) and a well-defined ligand structure could function as an effective laccase mimic for selectively catalyzing epinephrine (EP) oxidation. Catalytic activity of the CuZn-N/C nanozyme was superior to those of Cu-N/C and Zn-N/C and featured a Km value nearly 3-fold lower than that of natural laccase, which indicated that CuZn-N/C has a better affinity for EP. Density functional theory (DFT) revealed the mechanism of the superior catalytic ability of dual-metal CuZn-N/C as follows: (1) the exact distance of the two metal atoms in the CuZn-N/C catalyst makes it suitable for adsorption of the EP molecule, and the CuZn-N/C catalyst can offer the second hydrogen bond that stabilizes the adsorption; (2) molecular orbitals and density of states indicate that the strong interaction between the EP molecule and CuZn-N/C is important for EP catalytic oxidization. Furthermore, a sensitive and selective online optical detection platform (OODP) is constructed for determining EP with a low limit of detection (LOD) of 0.235 μM and a linear range of 0.2-20 μM. The system allows real-time measurement of EP release in the rat brain in vivo following ischemia with dexmedetomidine administration. This work not only provides an idea of designing efficient laccase mimics but also builds a promising chemical platform for better understanding EP-related drug action for ischemic cerebrovascular illnesses and opens up possibilities to explore brain function.

A nanoplatform-based aptasensor to electrochemically detect epinephrine produced by living cells.

Anovel aptasensor has beendesigned for quantitative monitoring of epinephrine (EP) based on cerium metal-organic framework (CeMOF) loaded gold nanoparticles (AuNPs). The aptamer, specific to EP, is immobilized on a flexible screen-printed electrode modified with AuNPs@CeMOF, enabling highly selective binding to the target biomolecule. Under optimized operational conditions, the peak currents using voltammetric detectionmeasured at voltage of 83mV (vs. Ag/AgCl) for epinephrine exhibit a linear increase within concentration in the range1pM-10nM. Following this detection strategy, a boasted limit of detection of 0.3pM was achieved, surpassing the sensitivity of most reported methods. The developed biosensor showcased exceptional performance in detection ofEP in spiked serum sample, with remarkable recovery range of 95.8-113% and precision reflected by low relative standard deviation (RSD) ranging from 2.23 to 6.19%. These results indicate the potential utility of this biosensor as a valuable clinical diagnostic tool. Furthermore, in vitro experiments were carried out using the presented biosensor to monitor the release of epinephrine from PC12 cells upon extracellular stimulation with K+ ions.

Comparison of anti-inflammatory influences of the splanchnic sympathetic nerve with those of the hypothalamic-pituitary-adrenal (HPA) axis during systemic inflammation

Branches of the splanchnic sympathetic nerve, that supply several abdominal organs including adrenals, provide the efferent neural pathway for reflex dampening of systemic inflammation (SI). In response to SI, the splanchnic nerves not only suppress secretion of the pro-inflammatory cytokine tumour necrosis factor-α (TNF) but drive release of the anti-inflammatory cytokine interleukin-10 (IL-10) (1,2,3). Recently we showed that the adrenal branch plays a major role in driving IL-10 secretion (3), while distal branches to other organs (e.g. spleen) suppress TNF secretion from immune cells (2,3). Glucocorticoid secretion resulting from activation of the HPA axis during SI is also anti-inflammatory. The present study compares the anti-inflammatory role of the adrenal cortex with that of the splanchnic nerve and adrenal medulla following i.v. lipopolysaccharide (LPS; 60 μg/kg) in urethane-anesthetized male rats. Plasma TNF and IL-10 levels sampled at 90 min after LPS injection were measured by ELISA. We hypothesized that the anti-inflammatory action of the splanchnic nerve differs from that of adrenal corticosteroids during SI. Gender differences were not investigated. We compared effects on endotoxin stimulated IL-10 and TNF secretion of cutting bilaterally (i) adrenal nerves, n=6, (ii) splanchnic nerves, n = 5, (iii) bilateral adrenalectomy, n = 8, (iv) combined adrenalectomy and splanchnic nerve cuts, n = 7, and (v) sham surgery, n = 5. As expected, TNF levels in response to LPS increased after cutting splanchnic (17.1 ± 2.5 ng/ml, mean ± SEM, p < 0.001) or adrenal (12.9 ± 3.8 ng/ml, p < 0.05) nerves compared with sham operated rats (3.9 ± 1.3 ng/ml). Adrenalectomy which reduces circulating adrenaline as well as corticosterone levels, resulted in greater plasma [TNF] (36.4 ± 5.2 ng/ml, p < 0.001), while combined splanchnic denervation and adrenalectomy further increased the TNF level to 61.3 ± 12. 4 ng/ml. Regarding plasma IL-10 levels, cutting either splanchnic or adrenal nerves alone reduced LPS-induced plasma IL-10 from 0.9 ± 0.2 ng/ml (sham) to 0.2 ± 0.1 and 0.3 ± 0.1 ng/ml respectively (both p < 0.05), whereas bilateral adrenalectomy alone (1.1 ± 0.1 ng/ml) or combined with splanchnic denervation (0.6 ± 0.1 ng/ml) did not. We conclude that glucocorticoids strongly suppress both pro- and anti-inflammatory cytokines. However, splanchnic sympathetic nerves are more selective, enhancing release of the anti-inflammatory cytokine IL-10, via adrenaline release, while suppressing pro-inflammatory cytokines such as TNF. References. (1) Martelli, D. et al., J Physiol. 592: 1677-86, 2014. (2) Martelli, D. et al., Am J Physiol Regul Integr Comp Physiol. 316: R235-242, 2019. (3) McKinley, M.J. et al., J Physiol. 600: 4521-36, 2022. NHMRC of Australia 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.

Nerve growth factor causes epinephrine release dysfunction by regulating phenotype alterations and the function of adrenal medullary chromaffin cells in mice with allergic rhinitis.

The presence of allergic rhinitis (AR) is an increased risk factor for the occurrence of bronchial asthma (BA). Nerve growth factor (NGF), in addition to its key role in the development and differentiation of neurons, may also be an important inflammatory factor in AR and BA. However, the pathogenesis of the progression of AR to BA remains to be elucidated. The present study aimed to investigate the ability of NGF to mediate nasobronchial interactions and explore possible underlying molecular mechanisms. In the present study, an AR mouse model was established and histology of nasal mucosa tissue injury was determined. The level of phenylethanolamine N‑methyl transferase in adrenal medulla was determined by immunofluorescence. Primary adrenal medullary chromaffin cells (AMCCs) were isolated and cultured from the adrenal medulla of mice. The expression levels of synaptophysin (SYP), STAT1, JAK1, p38 and ERK in NGF‑treated and untreated AMCCs were detected by reverse‑transcription‑quantitative PCR and western blotting. The epinephrine (EPI) and norepinephrine (NE) concentrations were measured by ELISA. It was found that the expression of SYP in AMCCs was enhanced in the presence of NGF, whereas, the concentration of EPI decreased significantly under the same conditions. Furthermore, NGF mediated the phenotypic and functional changes of AMCCs, resulting in decreased EPI secretion via JAK1/STAT1, p38 and ERK signaling. In conclusion, these findings could provide novel evidence for the role of NGF in regulating neuroendocrine mechanisms.

Relaxation of thoracic aorta and pulmonary artery rings of marmosets (Callithrix spp.) by endothelium-derived 6-nitrodopamine.

6-Nitrodopamine is a novel catecholamine released by vascular tissues, heart, and vas deferens. The aim of this study was to investigate whether 6-nitrodopamine is released from the thoracic aorta and pulmonary artery rings of marmosets (Callithrix spp.) and to evaluate the relaxing and anti-contractile actions of this catecholamine. Release of 6-nitrodopamine, dopamine, noradrenaline, and adrenaline was assessed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The relaxations induced by 6-nitrodopamine and by the selective dopamine D2 receptor antagonist L-741,626 were evaluated on U-46619 (3 nM)-pre-contracted vessels. The effects of 6-nitrodopamine and L-741,626 on the contractions induced by electric-field stimulation (EFS), dopamine, noradrenaline, and adrenaline were also investigated. Both aorta and pulmonary artery rings exhibited endothelium-dependent release of 6-nitrodopamine, which was significantly reduced by the NO synthesis inhibitor L-NAME. Addition of 6-nitrodopamine or L-741,626 caused concentration-dependent relaxations of both vascular tissues, which were almost abolished by endothelium removal, whereas L-NAME and the soluble guanylate cyclase inhibitor ODQ had no effect on 6-nitrodopamine-induced relaxations. Additionally, pre-incubation with 6-nitrodopamine antagonized the dopamine-induced contractions, without affecting the noradrenaline- and adrenaline-induced contractions. Pre-incubation with L-741,626 antagonized the contractions induced by all catecholamines. The EFS-induced contractions were significantly increased by L-NAME, but unaffected by ODQ. Immunohistochemical assays showed no immunostaining of the neural tissue markers S-100 and calretinin in either vascular tissue. The results indicated that 6-nitrodopamine is the major catecholamine released by marmoset vascular tissues, and it acts as a potent and selective antagonist of dopamine D2-like receptors. 6-nitrodopamine release may be the major mechanism by which NO causes vasodilatation.

Эффекты кардиотоксинов кобры на папиллярной мышце и сердце крысы, перфузируемом по Лангендорфу, не связаны с выбросом адреналина

In a study of the effects of cobra cardiotoxins on myocardial tissue, both right ventricular (papillary muscle) and left ventricular contractility (isovolumic recording of left ventricular pulse pressure during Langendorff perfusion of the heart) were assessed. In papillary muscle, both toxins, at a concentration of 5 μg/mL, caused short-term increases in contractility to 200±25% and 171±15% for CTX-1 and CTX-2, respectively, at the point of maximum effect. At the same time, for CTX-1 and CTX-2, the time to peak tension (TPT) increased from 104±2 to 111±2 and from 96±2 to 104±5 ms, the relaxation time to 50%(TR50%) from 64±4 to 70±6 and from 64±6 to 69±7 ms, and the relaxation time to 95%(TR95%) from 163±10 to 190±22 and from 148±16 to 155±20 ms, respectively. This significantly differs from the positive inotropic effect of the β-adrenomimetic isoproterenol (170±31%), which causes acceleration of TPT from 106±5 to 89±4 ms, TR50% from 58±6 to 43±4 ms, and TR95% from 145±15 to 90±14 ms. When the whole heart was exposed to cardiotoxins, an increase in contractility was also observed, followed by its suppression and contracture, in contrast to isoproterenol, which caused a steady increase in contractility coupled with an increase in heart rate. Pretreatment of papillary muscles with the β-blocker propranolol 
 (10 μM) did not prevent the development of cardiotoxin effects, but completely blocked the effects of isoproterenol. Our data indicate that the temporary increase in contractility under the action of cardiotoxins is not associated with the release of endogenous adrenaline, but rather is caused by changes in calcium homeostasis in cardiomyocytes.

ODP291 A Series of COVID Reinfections in a Vaccinated Patient leading to Development of Neurogenic Orthostatic Hypotension and Postural Orthostatic Tachycardia

Abstract The recently recognized 'long COVID' syndrome, encompasses symptoms of shortness of breath, chest pain, palpitations and orthostatic intolerance which can last for weeks or more following even mild illness. 'Long COVID' is postulated to be related to a virus- or immune-mediated disruption of the autonomic nervous system resulting in orthostatic intolerance syndromes. We report a 32-yr old female nurse who presented to the endocrinology clinic for symptoms of orthostatic hypotension following two COVID 19 infections. She had received 1 dose of Moderna vaccine in February and March of 2021 respectively. She was hospitalized for COVID June and September of 2021. Following the first hospitalization, she developed severe orthostatic hypotension, chest pain, shortness of breath, fatigue, anxiety, memory, concentration and word finding difficulty. Blood pressure ranged from 110/78 to 124/82 while lying and from 85/45 to 103/59 while standing. HR ranged from 88 to 96 while lying and from 130 to 42 while standing. She was started on midodrine 10 mg three times daily which was discontinued due to patient intolerance. She followed a regimen of adequate hydration, adequate salt intake, fall precautions, physical and speech therapy and high protein/low carb diet with frequent snacks, which improved her symptoms of dizziness and orthostasis, but without complete resolution. However, symptoms of anxiety, dizziness, chest pain and palpitations became even worse after the second hospitalization. She was started on Propranolol 20 mg 2-3 times daily depending on symptoms, and she felt better with this intervention. Evaluation for secondary causes of hypotension were all negative [Cortisol of 19 ug/dL (n=5.3-22.5); ACTH 20 pg/mL (n=0-47); TSH 2.36 uIU/mL (n=0.358 -3.74); Free T4 0.97 ng/dL (n=0.76-1.46); B12 480 pg/mL (n=193-986); Hba1c 4.9%; 25 OH Vitamin D 24.3 ng/mL; tryptase 3.7 mcg/L (n= <11); Sed rate 4 mm/HR (n=0-20). CBC, CMP, TPO and Tg abs, troponin levels, iron studies, ANA panel, Rheumatoid factor, anti-CCP abs, myasthenia gravis panel], CT head and MRI brain, EKG and QTc interval. Echocardiogram= small PFO. 'Post-acute COVID' refers to persistent symptoms 3 weeks after COVID-19 infection, while 'Chronic COVID' describes symptoms lasting more than 12 weeks. Symptoms include fatigue, dyspnea, chest pain, palpitations, dizziness, body aches, presyncope and orthostatic intolerance. Individuals may even develop post-traumatic stress disorder, panic attacks and irritable bowel syndrome. Orthostatic intolerance occurs due to release of epinephrine and norepinephrine leading to development of palpitations, breathlessness, and chest pain. It has been well described that orthostatic intolerance is preceded by viral infections and is associated with the development of autoantibodies α-/β-adrenoceptors and muscarinic receptors. COVID 19 itself can affect the autonomic nervous system through the release of a cytokine storm leading to sympathetic activation. It is crucial to consider the diagnosis of orthostatic intolerance syndromes post COVID infection. Presentation: No date and time listed

PSIII-A-7 Conditioning of Goats to Livestock Trailer: Effects on Physiological Stress Responses During Long-Distance Transportation

Abstract Meat goats are often transported long distances under commercial situations in the US. In a split-plot design, 168 twelve-month-old male Spanish goats were used to determine the effects of habituation to trailers on behavior and stress responses when transported for long periods. Animals were separated into two treatment (TRT) groups and maintained on two different paddocks. Concentrate supplement was fed to one group inside two livestock trailers (5.0 × 2.3 m each; habituated group, H), while the other group received the same quantity of concentrate, but not inside the trailers (non-habituated, NH). After 4 weeks of habituation period, goats were subjected to a 10-hour transportation stress in 4 replicates (n = 21 goats/replicate/TRT). Blood samples were collected prior to loading (Preload), 20 min after loading (0 h), and at 2, 4, 6, 8, and 10 h of transportation (Time). Behaviors of goats after loading were not affected by treatment. Plasma cortisol, glucose, and urea nitrogen concentrations and creatine kinase (CK) activities were significantly influenced by Time (P < 0.01). Plasma cortisol was not affected by TRT; however, glucose and CK in the H group tended to be lower than the NH group. Plasma non-esterified fatty acid (NEFA) concentrations were significantly lower (P < 0.01) in the H group than the NH group, likely in response to epinephrine release. There was a TRT × Time effect (P < 0.05) on CK activities, with the values increasing in the NH group and decreasing in the H group during the first two hours of transportation. Lymphocyte to neutrophil ratio also increased (P < 0.01) over transportation time. Habituation to trailer may be beneficial during long distance transportation of goats as indicated by certain physiological stress responses. Habituation may also facilitate the loading process, based on behavioral observations.

Molecular regulation, breed differences and genes involved in stress control in farm animals

Stress is a state of disturbed homeostasis evoking a multiplicity of somatic and mental adaptive reactions resulting from any of the 5 freedoms of animals being violated. Many environmental forces disrupt homeostasis in farm animals, such as extreme temperatures, poor nutrition, noise, hunger, and thirst. During stressful situations, neuronal circuits in the limbic system and prefrontal cortex are activated, which lead to the release of adrenalin and noradrenalin. The hormones released during stress are needed for adaptation to acute stress and are regulated by many genes. This review examined molecular regulation, breed differences, and genes involved in stress control in farm animals. Major molecular regulation of stress, such as oxidative, cytosolic heat shock, unfolded protein, and hypoxic responses, were discussed. The responses of various poultry, ruminant, and pig breeds to different stress types were also discussed. Gene expressions and polymorphisms in the neuroendocrine and neurotransmitter pathways were also elucidated. The information obtained from this review will help farmers mitigate stress in farm animals through appropriate breed and gene-assisted selection. Also, information obtained from this review will add to the field of stress genetics since stress is a serious welfare issue in farm animals.