Beta-adrenergic Agonist Research Articles

Ractopamine-induced changes in sarcoplasmic proteome profile of post-rigor pork <i>semimembranosus</i> muscle

Ractopamine is a beta-adrenergic agonist that increases leanness and carcass weight in finishing pigs. Our previous study observed that dietary ractopamine increased the abundance of several glycolytic enzymes in the sarcoplasmic proteome of post-rigor pork longissimus thoracis muscle. Pork semimembranosus is an economically important muscle and demonstrates differences in biochemistry compared with longissimus thoracis. Nonetheless, the effects of ractopamine on sarcoplasmic proteome of semimembranosus have not been evaluated yet. Therefore, this study examined the influence of ractopamine on sarcoplasmic proteome of post-rigor pork semimembranosus. Analyses of sarcoplasmic proteome of semimembranosus muscles from control (CON; diet without ractopamine) and ractopamine-fed (RAC; 7.4 mg/kg for 14 days followed by 10.0 mg/kg for 14 days) barrows revealed that haemoglobin subunit beta, alpha-crystallin B, and titin fragments were over-abundant in CON. In contrast, myosin light chain 1/3 and tripartite motif-containing protein 72 were over-abundant in RAC. The low abundance of haemoglobin subunit beta and alpha crystallin B in RAC could be attributed to fibre type shift (from oxidative to glycolytic) in response to ractopamine. The over-abundance of MLC 1/3 and tripartite motif-containing protein 72 in RAC could be due to the increased myofibrillar protein synthesis and muscle mass in ractopamine-fed pigs. Dietary ractopamine decreased the abundance of sarcoplasmic proteins involved in oxygen transport and chaperone activity, but increased the abundance of proteins involved in muscle contraction and plasma membrane repair in pork semimembranosus muscle. Keywords : Pork, ractopamine, sarcoplasmic proteome, semimembranosus

The Insular Cortex Dynamically Maps Changes in Cardiorespiratory Interoception

Palpitations and dyspnea are fundamental to the human experience of panic anxiety, but it remains unclear how the brain dynamically represents changes in these interoceptive sensations. We used isoproterenol, a rapidly acting peripheral beta-adrenergic agonist similar to adrenaline, to induce sensations of palpitation and dyspnea in healthy individuals (n=23) during arterial spin labeling functional magnetic resonance imaging (fMRI). We hypothesized that the right mid-insular cortex, a central recipient of viscerosensory input, would preferentially respond during the peak period of cardiorespiratory stimulation. Bolus infusions of saline and isoproterenol (1 or 2 μg) were administered in a blinded manner while participants continuously rated the intensity of their cardiorespiratory sensation using a dial. Isoproterenol elicited dose-dependent increases in cardiorespiratory sensation, with all participants reporting palpitations and dyspnea at the 2 μg dose. Consistent with our hypothesis, the right mid-insula was maximally responsive during the peak period of sympathetic arousal, heart rate increase, and cardiorespiratory sensation. Furthermore, a shift in insula activity occurred during the recovery period, after the heart rate had largely returned to baseline levels, with an expansion of activation into anterior and posterior sectors of the right insula, as well as bilateral regions of the mid-insula. These results confirm the right mid-insula is a key node in the interoceptive network, and inform computational models proposing specific processing roles for insula subregions during homeostatic inference. The combination of isoproterenol and fMRI offers a powerful approach for evaluating insula function, and could be a useful probe for examining interoceptive dysfunction in psychiatric disorders.

Trends in Severe Obesity Among 23 Million U.S. Children Aged 2–4 Years Who Enrolled in WIC — United States, 2000–2014

The aim of this study was to verify β2-AR expression in oral squamous cell carcinoma cell lines (SCC-9 and SCC-25), and to investigate the role of this receptor in migration and invasion of these neoplastic cells.SCC-9 and SCC-25 cells were investigated for gene and protein expression of β2-AR. Cell migration and invasion were analyzed by wound healing assay and transwell invasion camera system. Different concentrations (0.1, 1 and 10 μM) of norepinephrine were used to stimulate, and 1 μM propranolol was used to block the beta-adrenergic receptors on cancer cells. Differences in median values of SCC-9 and SCC-25 and β2-AR protein expression were analyzed by Friedman test and in case of significant differences; pairwise comparisons were performed using Bonferroni correction.The results showed that the β2-AR gene and protein expression were observed in both oral cancer cell lines. The concentration of 10 μM of norepinephrine significantly inhibited (p ≤ 0.05) migration of SCC-9 and SCC-25 cell lines. Furthermore, there was a significant reduction (p ≤ 0.05) in the effect of norepinephrine on cell migration when the β2-AR was inhibited by propranolol. The blockade by propranolol showed a tendency to reverse the effect of norepinephrine on the invasiveness of SCC-9 and SCC-25.The use of beta-adrenergic receptor agonists could become an adjuvant therapeutic target in the treatment of this malignancy.

A3 Early Career Award Speaker: Interactions between trace minerals and growth promoting practices in beef cattle

Trace minerals (TM) are necessary to support optimum animal growth. While TM requirements to prevent deficiency were recently updated, the requirements of beef cattle to support optimal performance remain ill defined. Advancements in animal genetics and utilization of technologies to promote growth of feedlot cattle have dramatically advanced production efficiencies in the beef industry. Trace minerals (Co, Cu, Fe, I, Mn, Se and Zn) are integral in a variety of growth processes in the body, including supporting metalloproteins involved in DNA synthesis, collagen production, energy utilization and critical antioxidant enzymes. Over 80% of feedlot cattle are estimated to receive hormone implants to enhance growth and efficiency. Recently, we examined the effect of feeding no supplemental TM, TM supplemented at NRC recommendations, or TM supplemented at industry recommended concentrations (all inorganic sources), fed to Angus cross steers receiving no implant or implanted with Component TE-IS, followed by Component TE-200 on d 56. Harvested on d 124, HCW was, as expected, improved (P = 0.0001) by implant, but HCW was also improved by 15 kg (P = 0.01) in steers supplemented at industry TM concentrations vs. those receiving no supplemental TM. Additional work is needed to clarify the potentially greater need for TM in cattle receiving hormone implants. Introduction of beta adrenergic agonists revolutionized late stage finishing cattle performance, increasing feed efficiency, ADG, and often HCW of cattle. In a series of studies we have found that increasing supplementation of Zn (from a combination of inorganic and organic sources) to steers fed ractopamine hydrochloride increases cattle performance (ADG, feed efficiency, and HCW in some studies) during the ractopamine feeding period. Zinc is thought to inhibit breakdown of the intracellular signalling molecule, cAMP, perhaps potentiating the growth response. Alternately, demand for Zn and other TM may simply be greater due to increased growth rates. We recently found the retention of Zn and N to be positively correlated and that both ractopamine hydrochloride and Zn increase N retention in steers. It seems likely that growth enhancing technologies influence TM requirements of cattle, and further work is needed to determine TM requirements that optimize cattle performance and maximize producer profitability.

Dynamic transcriptome changes during adipose tissue energy expenditure reveal critical roles for long noncoding RNA regulators.

Enhancing brown fat activity and promoting white fat browning are attractive therapeutic strategies for treating obesity and associated metabolic disorders. To provide a comprehensive picture of the gene regulatory network in these processes, we conducted a series of transcriptome studies by RNA sequencing (RNA-seq) and quantified the mRNA and long noncoding RNA (lncRNA) changes during white fat browning (chronic cold exposure, beta-adrenergic agonist treatment, and intense exercise) and brown fat activation or inactivation (acute cold exposure or thermoneutrality, respectively). mRNA–lncRNA coexpression networks revealed dynamically regulated lncRNAs to be largely embedded in nutrient and energy metabolism pathways. We identified a brown adipose tissue–enriched lncRNA, lncBATE10, that was governed by the cAMP-cAMP response element-binding protein (Creb) axis and required for a full brown fat differentiation and white fat browning program. Mechanistically, lncBATE10 can decoy Celf1 from Pgc1α, thereby protecting Pgc1α mRNA from repression by Celf1. Together, these studies provide a comprehensive data framework to interrogate the transcriptomic changes accompanying energy homeostasis transition in adipose tissue.

743 Using advanced technologies to quantify cattle behavior

For many decades, we have relied upon visual observation of animal behavior to define clinical disease, assist in breeding selection and predict growth performance. Limitations of visual monitoring of cattle behavior include training of personnel, subjectivity, and brevity; however, extensive time and labor is required to monitor behavior in a large number of animals. Furthermore, the prey instinct of cattle to disguise abnormal behaviors in the presence of a human evaluator is problematic. More recently, cattle behavior has been quantified objectively and continuously using advanced technologies to assess animal welfare, indicate lameness or disease, and detect estrus in both production and research settings. Some of these technologies include 3-axis accelerometers that quantify physical activity, systems that document feeding and watering behavior via radio frequency, and triangulation or global positioning systems to determine location and movement of cattle within a pen or pasture. Advances in Wi-Fi and radio frequency technology have allowed many of these systems to operate in real-time and efforts are underway to develop commercial applications that may allow early detection of respiratory or other cattle diseases in the production environment. Current challenges with commercial application of technology for early disease detection include establishment of an appropriate algorithm to ensure maximum sensitivity and specificity, cost benefit, and infringement of traditional methodology for clinical diagnosis. Advanced technologies have also allowed researchers to determine temporal variance in behavior or variability between experimental treatments. Specifically, we have used 3-axis accelerometers to continuously record standing and lying duration, step count, lying bouts and motion index generated in 15 min intervals. Our research efforts include effects of castration method and timing, weaning method, transportation, beta-adrenergic agonist feeding, and bovine respiratory disease on physical behavior in cattle. Benchmark activity data will also be discussed for cattle fed in confinement and during transport and lairage at a commercial beef processor.

Discovery of novel brain permeable and G protein-biased beta-1 adrenergic receptor partial agonists for the treatment of neurocognitive disorders.

The beta-1 adrenergic receptor (ADRB1) is a promising therapeutic target intrinsically involved in the cognitive deficits and pathological features associated with Alzheimer’s disease (AD). Evidence indicates that ADRB1 plays an important role in regulating neuroinflammatory processes, and activation of ADRB1 may produce neuroprotective effects in neuroinflammatory diseases. Novel small molecule modulators of ADRB1, engineered to be highly brain permeable and functionally selective for the G protein with partial agonistic activity, could have tremendous value both as pharmacological tools and potential lead molecules for further preclinical development. The present study describes our ongoing efforts toward the discovery of functionally selective partial agonists of ADRB1 that have potential therapeutic value for AD and neuroinflammatory disorders, which has led to the identification of the molecule STD-101-D1. As a functionally selective agonist of ADRB1, STD-101-D1 produces partial agonistic activity on G protein signaling with an EC50 value in the low nanomolar range, but engages very little beta-arrestin recruitment compared to the unbiased agonist isoproterenol. STD-101-D1 also inhibits the tumor necrosis factor α (TNFα) response induced by lipopolysaccharide (LPS) both in vitro and in vivo, and shows high brain penetration. Other than the therapeutic role, this newly identified, functionally selective, partial agonist of ADRB1 is an invaluable research tool to study mechanisms of G protein-coupled receptor signal transduction.

Effectiveness and safety of the oxytocin antagonist( atosiban) versus beta-adrenergic agonists (salbutamol) in the treatment of preterm labor

Background and Objectives: preterm labor complicates 5-10% of pregnancies and is a leading cause of neonatal morbidity and mortality worldwide and 70-80% of perinatal deaths occur in preterm infants. The aim of this study is to compare the effectiveness, safety and adverse effects of the oxytocin antagonist medication(atosiban) with those of beta-adrenergic agonist (salbutamol) in the treatment of patients with preterm labor.Patients and Methods: one hundred pregnant women with preterm labor were enrolled in this study from the period of( January 2014 – January 2015) at Al-Diwaniya Maternity and Pediatrics Teaching Hospital-Iraq with a gestational age of 24-34 weeks, they were randomly assigned to receive tocolytics either salbutamol (n=50) or atosiban (n=50).Salbutamol was given by(intravenous infusion 10-50 microgram)for up to 48 hour. Atosiban was given by (intravenous bolus dose of 6.75 mg then 300microgram/minute for 3 hour and 100microgram/minute for up to 48hour). Retreatment with the study drugs or alternative tocolytic agents was allowed. Main outcome measures included were tocolytic effectiveness, which was assessed in terms of number of women undelivered after 48hour and 7 days. Tocolytic safety was assessed in terms of maternal and fetal side effects and neonatal morbidity.Results: there were no significant differences between the salbutamol and the atosiban group in prolongation of pregnancy for 48 hour (44% versus 46%;p=0841)and 7 days(20% versus 32%;p=0.171) ,respectively. Maternal adverse events , including tachycardia occurred more frequently in the salbutamol group(22% versus 8%;p=0.050).Neonatal outcomes and complications were comparable between the two study groups(42% versus 32%;p=0.30).Conclusions: the oxytocin antagonist (atosiban) was as effective as beta-agonist(salbutamol) in delaying threatened preterm birth , and found to be better tolerated by both the mother and fetus than salbutamol , with a comparable neonatal safety profil.This study supports the clinical use of atosiban as a first line tocolytic in the treatment of preterm labor.

Abstract 324: Identification of Regulators of Cardiac DNA Methylation Using a Systems Genetics Approach in Mice

Systems biology approaches to studying heart failure in humans are hampered by numerous environmental factors and inciting incidents which combine to complicate the application of standard population-based tools and algorithms to the syndrome. We recently reported a study of heart failure in a large cohort of inbred mouse strains (the Hybrid Mouse Diversity Panel; HMDP) treated with the beta-adrenergic agonist isoproterenol. We demonstrated that it was possible to combine systems-level tools with the benefits of an animal model to capture and identify numerous genetic variations which drive heart failure. Recent research on the epigenome has identified changes to DNA methylation which affect the progression of cardiovascular diseases and suggests that DNA methylation may serve as an important additional marker or even driver of genes involved in heart failure-associated phenotypes. We have supplemented our prior study by completing reduced representational bisulfite sequencing on 88 strains from our panel both with and without catecholamine challenge. We have identified 75,000 CpGs which vary among the strains of the HMDP across the genome. Each CpG was mapped to the genome using GWAS to identify methylation quantitative trait loci (mQTLs). These mQTLs were then grouped together to identify mQTL hotspots, regions on the genome which regulate a sizable fractions of all observed CpGs. We then overlapped these hotspots with hotspots which regulate the expression of gene transcripts. Three genomic regions were identified as multi-omic master regulators which affect both DNA methylation and gene expression for over 5% of the expressed transcripts and varying CpGs. Closer examination of these loci identified the serine peptidase inhibitor Serpina3n as a likely candidate gene for the regulation of DNA methylation, gene expression and cardiac phenotypes. Subsequent in vitro and in vivo work demonstrate that Serpina3n knockdown results in reduced cellular hypertrophy and changes to hypertrophy-related gene expression. Further analysis of the other master regulatory regions will likely reveal additional genes which will improve our understanding of how DNA methylation acts to regulate pathways underlying heart failure.

The neuropeptide galanin promotes an anti-thrombotic phenotype on endocardial endothelial cells from heart failure patients.

Thromboembolic complications are a significant cause of mortality and re-hospitalization in heart failure (HF) patients. One source of thrombi is the ventricular endocardial surface that becomes increasingly pro-thrombotic as HF progresses. Anticoagulation comes with bleeding risks so identifying therapeutic agents for improving cardiac endothelial health are of critical clinical importance. Endocardial endothelial cells are closely apposed to cardiac sympathetic nerves. In HF, cardiac sympathetic nerves are dysregulated and promote disease progression. Whether endocardial endothelial health and function is impacted by sympathetic dysregulation in HF is unknown. Also unexplored is the impact of neuropeptides, such as galanin and neuropeptide Y (NPY), co-released from sympathetic nerve terminals, on endothelial health. In this study we examined the effect of sympathetic nerve-released neurotransmitters and neuropeptides on the procoagulant phenotype of cultured human endocardial endothelial cells from HF patients. As a functional readout of procoagulant state we examined thrombin-mediated von Willebrand factor (vWF) extrusion and multimer expression. We demonstrate that vWF extrusion and multimer expression is promoted by thrombin, that isoproterenol (a beta-adrenergic receptor agonist) augments this effect, whereas co-treatment with the beta-blockers propranolol and carvedilol blocks this effect. We also show that vWF extrusion and multimer expression is attenuated by treatment with the neuropeptide galanin, but not with NPY. Our results are consistent with a protective role of beta-blockers and galanin on endocardial endothelial health in heart failure. Improving endothelial health through galanin therapy is a future clinical application of this study.

Engineered 3D Cardiac Fibrotic Tissue to Study Fibrotic Remodeling.

Activation of cardiac fibroblasts into myofibroblasts is considered to play an essential role in cardiac remodeling and fibrosis. A limiting factor in studying this process is the spontaneous activation of cardiac fibroblasts when cultured on two-dimensional (2D) culture plates. In this study, a simplified three-dimensional (3D) hydrogel platform of contractile cardiac tissue, stimulated by transforming growth factor-β1 (TGF-β1), is presented to recapitulate a fibrogenic microenvironment. It is hypothesized that the quiescent state of cardiac fibroblasts can be maintained by mimicking the mechanical stiffness of native heart tissue. To test this hypothesis, a 3D cell culture model consisting of cardiomyocytes and cardiac fibroblasts encapsulated within a mechanically engineered gelatin methacryloyl hydrogel, is developed. The study shows that cardiac fibroblasts maintain their quiescent phenotype in mechanically tuned hydrogels. Additionally, treatment with a beta-adrenergic agonist increases beating frequency, demonstrating physiologic-like behavior of the heart constructs. Subsequently, quiescent cardiac fibroblasts within the constructs are activated by the exogenous addition of TGF-β1. The expression of fibrotic protein markers (and the functional changes in mechanical stiffness) in the fibrotic-like tissues are analyzed to validate the model. Overall, this 3D engineered culture model of contractile cardiac tissue enables controlled activation of cardiac fibroblasts, demonstrating the usability of this platform to study fibrotic remodeling.

Accidental Intramuscular Isoprenaline in Early Pregnancy: The Effects, Management and Outcome

Isoprenaline is a beta-adrenergic agonist, used for the treatment of cardiovascular emergencies like hypotensive shock or sever bradycardia. This drug is not indicated to use in bolus form either intramuscularly or intravenously. Accidentally a 22 year old pregnant woman was given an intramuscular injection of two mg Isoprenaline. It was a lethal dose, fortunately the patient survived. This article narrated the events, the ECG changes, management and outcome of the patient and pregnancy.J Bangladesh Coll Phys Surg 2016; 34(2): 108-111

Translational science approach for assessment of cardiovascular effects and proarrhythmogenic potential of the beta-3 adrenergic agonist mirabegron

IntroductionTranslational assessment of cardiac safety parameters is a challenge in clinical development of beta-3 adrenoceptor agonists. The preclinical tools are presented that were used for assessing human safety for mirabegron. MethodsStudies were performed on electrical conductance at ion channels responsible for cardiac repolarization (IKr, IKs, Ito, INa, and ICa,L), on QT-interval, subendocardial APD90, Tpeak-end interval, and arrhythmia's in ventricular dog wedge tissue in vitro and on cardiovascular function (BP, HR, and QTc) in conscious dogs. ResultsIn conscious dogs, mirabegron (0.01–10mg/kg, p.o.) dose-dependently increased HR, reduced SBP but DBP was unchanged. Propranolol blocked the decrease in SBP and attenuated HR increase at 100mg/kg mirabegron. Mirabegron, at 30, 60, or 100mg/kg, p.o., had no significant effect on the QTc interval. In paced dog ventricular wedge, neither mirabegron nor metabolites M5, M11, M12, M14, and M16 prolonged QT, altered transmural dispersion of repolarization, induced premature ventricular contractions, or induced ventricular tachycardia. Mirabegron nor its metabolites inhibited IKr, IKs, Ito INa, or ICa,L at clinically relevant concentrations. DiscussionUp to exposure levels well exceeding human clinical exposure no discernible effects on ion channel conductance or on arrhythmogenic parameters in ventricular wedge resulted for mirabegron, or its main metabolites, confirming human cardiac safety findings. In vivo, dose-related increases in HR with effects markedly higher than seen clinically, was mediated in part by cross-activation of beta-1 adrenoceptors. This non-clinical cardiac safety test program therefore proved predictive for human cardiac safety for mirabegron.

Beta Adrenergic Regulation of Intrapulmonary Arteriovenous Anastomoses in Intact Rat and Isolated Rat Lungs

Intrapulmonary arteriovenous anastomoses (IPAVA) allow large diameter particles of venous origin to bypass the pulmonary capillary bed and embolize the systemic arterial circulation. IPAVA have been routinely observed in healthy humans with exercise, hypoxia, and catecholamine infusion, but the mechanism by which they are recruited is not well-defined. We hypothesized that beta-adrenergic receptor stimulation recruits IPAVA and that receptor blockade would limit hypoxia-induced IPAVA recruitment. To test our hypothesis, we evaluated the transpulmonary passage of microspheres in intact rats and isolated rats lung infused with the beta-adrenergic receptor agonist isoproterenol. We also evaluated IPAVA recruitment in intact rats with hypoxia and the beta-adrenergic receptor blocker propranolol. We found that IPAVA are recruited in the intact rat by isoproterenol and their recruitment by hypoxia can be minimized by propranolol, suggesting a role for the adrenergic system in the recruitment of IPAVA by hypoxia. IPAVA recruitment is completely abolished by ventilation with 100% oxygen. Isoproterenol also recruits IPAVA in isolated rat lungs. The fact that isoproterenol can recruit IPAVA in isolated lungs, without increased pulmonary flow, suggests that elevated cardiac output is not required for IPAVA recruitment.

Dialkyl anionic surfactant in field-amplified sample injection and sweeping-micellar electrokinetic chromatography for determination of eight leanness-promoting β-agonists in animal feeds

The beta-adrenergic agonists (β-agonists) working as repartitioning agents that make the carcass leaner and enhance the feeding efficiency in animals have been banned in the European Union, China and Taiwan. Here, traditional anionic surfactants, such as sodium dodecyl sulfate (SDS) were replaced with sodium di-(2-ethylhexyl)-sulfosuccinate (AOT) in field-amplified sample injection and sweeping-micellar electrokinetic chromatography (FASI-sweeping MEKC) for simultaneous analysis of eight β-agonists in animal feeds. The AOT vesicles provided a better resolution of β-agonists than micelles of SDS. The detection limits of the eight β-agonists were above 5ng/mL by using this stacking capillary electrophoresis (CE) method. In comparison of traditional MEKC method (sample injection, 1psi for 5s), the stacking strategy provided 400–2000 fold sensitivity enhancement. After method validation, this method was successfully applied for analyzing four animal feeds, and none β-agonist was detected. This strategy possessing good resolution of eight β-agonists was suitable for serving as a tool for routine analysis of animal feeds.

Beta 2-adrenergic receptor agonists are novel regulators of macrophage activation in diabetic renal and cardiovascular complications

Macrophage activation is increased in diabetes and correlated with the onset and progression of vascular complications. To identify drugs that could inhibit macrophage activation, we developed a cell-based assay and screened a 1,040 compound library for anti-inflammatory effects. Beta2-adrenergic receptor (β2AR) agonists were identified as the most potent inhibitors of phorbol myristate acetate-induced tumor necrosis factor-α production in rat bone marrow macrophages. In peripheral blood mononuclear cells isolated from streptozotocin-induced diabetic rats, β2AR agonists inhibited diabetes-induced tumor necrosis factor-α production, which was prevented by co-treatment with a selective β2AR blocker. To clarify the underlying mechanisms, THP-1 cells and bone marrow macrophages were exposed to high glucose. High glucose reduced β-arrestin2, a negative regulator of NF-κB activation, and its interaction with IκBα. This subsequently enhanced phosphorylation of IκBα and activation of NF-κB. The β2AR agonists enhanced β-arrestin2 and its interaction with IκBα, leading to downregulation of NF-κB. A siRNA specific for β-arrestin2 reversed β2AR agonist-mediated inhibition of NF-κB activation and inflammatory cytokine production. Treatment of Zucker diabetic fatty rats with a β2AR agonist for 12 weeks attenuated monocyte activation as well as pro-inflammatory and pro-fibrotic responses in the kidneys and heart. Thus, β2AR agonists might have protective effects against diabetic renal and cardiovascular complications.

A case report of patient who had two manic episodes with psychotic features induced by nasal decongestant

ObjectivePhenylephrine, pseudoephedrine and ephedrine are the sympathomimetic drugs that have been used most commonly in oral preparations for the relief of nasal congestion. These drugs stimulate the central nervous system that is affected by the alpha and beta adrenergic agonism. Sympathomimetic agents used in the treatment of flu and common cold with ephedrine and pseudoephedrine are case reports. That the manic and psychotic episodes are triggered. In this article, we would like to present a bipolar manic disorder with two manic episodes and both of them triggered by influenza drugs.CaseA 25-year-old man patient was admitted to psychiatric outpatient clinic with increasing complaints such as increasing energy, speaking much, decreasing sleep, increasing the libido after using the flu drug that prescribed to him containing phenylephrine. Also, 2 years ago, he has manic attack triggered after the flu drug.DiscussionIn recent years the study of epilepsy and bipolar disorder in common suggests that bipolar disorder may affect the kindling phenomenon. In our case, two of reasons in the neurobiology of developing manic attacks the kindling phenomenon is likely to be effective. First, the possibility of using cold medicine containing ephedrine or pseudoephedrine in the first manic episode, in the second manic episode having spent the attack with FAQ stimulant effect of lower phenylephrine. Second, in the first episode after using the 5–6 tablets developing manic attacks. In the second attack to be triggered with just 2 doses may indicate the effect of kindling.Disclosure of interestThe authors have not supplied their declaration of competing interest.

Hyperspectral Measurements Allow Separation of FRET Signals from Non-Uniform Background Fluorescence

In recent years Forster resonance energy transfer (FRET) has become a standard imaging approach to gain insight into localized biochemical processes within cells. These processes include changes in protein colocalization, second messenger concentration, and activation of effector proteins such as protein kinase A (PKA). However, there are several limitations in FRET measurements that are not often considered. In recent years our group has used hyperspectral imaging approaches to address a subset of these limitations, including the inherently low signal-to-noise ratio of standard two and three filter set FRET measurements. Here we present data demonstrating that hyperspectral imaging approaches can be used to correct for non-uniform background fluorescence in low intensity FRET measurements. We utilized the FRET-based cAMP probe H188. The H188 probe contains a cAMP binding domain between donor (Turquoise) and acceptor (Venus) fluorescent proteins. The probe was transfected into pulmonary microvascular endothelial cells plated on glass coverslips. Hyperspectral image stacks were acquired using a Nikon A1R inverted confocal microscope. Changes in cAMP were triggered by addition of 0.1 µM isoproterenol (a beta adrenergic receptor agonist) 0.1 µM PGE1 (a prostanoid receptor agonist) or 50 µM forskolin (an adenylyl cyclase activator). Data were analyzed using Nikon Elements software and custom MATLAB scripts. Results demonstrate that non-uniform background fluorescence associated with glass coverslips can contaminate FRET measurements from weak fluorescence signals. Linear unmixing approaches were able to separate the abundances of background, donor and acceptor fluorescence signals. Thus, results from this study demonstrate that hyperspectral unmixing approaches can readily separate non-uniform background fluorescence from other fluorescence signatures, allowing for more accurate quantification of FRET efficiency. This work was supported by NIH P01HL066299, NIH S10RR027535, NIH T32HL076125, AHA 16PRE27130004, USA SURF Program, and the Abraham Mitchell Cancer Research Fund.

Glucokinase Mediated Glucosensing in Hypothalamic Neurons

Glucosensing is the ability of specialized cells to detect changes in extracellular glucose concentration and convert this information into a change in membrane potential; this allows glucose levels to influence the activity of cells that regulate glucose-altering or glucose-dependent physiological processes. Two major glucosensing cellular populations are pancreatic and hypothalamic cells; the mechanism of glucosensing in pancreatic beta cells is established, but that of hypothalamic neurons remains controversial. Beta cells glucosense by expressing a specialized type of hexokinase (the rate-limiting enzyme in glucose metabolism) called glucokinase (GCK). GCK, unlike standard hexokinase, is not saturated at physiological levels of glucose, allowing glucose metabolism in cells expressing GCK to increase when the extracellular glucose concentration is raised. This increase in metabolism is then converted into a membrane depolarization. Furthermore, S-nitrosylation can enhance the activity of GCK, and therefore enhance glucose metabolism and glucosensing. Here we use GT1-7 cells to explore whether a similar mechanism of glucosensing exists in hypothalamic neurons. The addition of a glucose concentration at which GCK is sensitive (and standard hexokinase is saturated) leads to an increase in cellular metabolism, as measured by NADH autofluorescence; but only in the presence of isoproterenol, a beta-adrenergic receptor agonist which increases intracellular calcium to activate neuronal nitric oxide synthase and facilitate S-nitrosylation. We also use a FRET-based GCK sensor to measure GCK activation under low and high glucose concentrations and in the presence of isoproterenol. These studies reveal the presence of a functional GCK and the sensitivity of GCK to post-translational/signaling cascade modulation. Further, the lack of metabolic response to glucose alone may be linked to high basal activity of GCK in this serum-grown cell line. These results point to a functional, receptor-potentiated glucose sensing mechanism in neurons that is mediated through post-translational activation of GCK.

Modulation of neuroinflammation and pathology in the 5XFAD mouse model of Alzheimer's disease using a biased and selective beta-1 adrenergic receptor partial agonist

Degeneration of noradrenergic neurons occurs at an early stage of Alzheimer's Disease (AD). The noradrenergic system regulates arousal and learning and memory, and has been implicated in regulating neuroinflammation. Loss of noradrenergic tone may underlie AD progression at many levels. We have previously shown that acute administration of a partial agonist of the beta-1 adrenergic receptor (ADRB1), xamoterol, restores behavioral deficits in a mouse model of AD. The current studies examined the effects of chronic low dose xamoterol on neuroinflammation, pathology, and behavior in the pathologically aggressive 5XFAD transgenic mouse model of AD. In vitro experiments in cells expressing human beta adrenergic receptors demonstrate that xamoterol is highly selective for ADRB1 and functionally biased for the cAMP over the β-arrestin pathway. Data demonstrate ADRB1-mediated attenuation of TNF-α production with xamoterol in primary rat microglia culture following LPS challenge. Finally, two independent cohorts of 5XFAD and control mice were administered xamoterol from approximately 4.0–6.5 or 7.0–9.5 months, were tested in an array of behavioral tasks, and brains were examined for evidence of neuroinflammation, and amyloid beta and tau pathology. Xamoterol reduced mRNA expression of neuroinflammatory markers (Iba1, CD74, CD14 and TGFβ) and immunohistochemical evidence for microgliosis and astrogliosis. Xamoterol reduced amyloid beta and tau pathology as measured by regional immunohistochemistry. Behavioral deficits were not observed for 5XFAD mice. In conclusion, chronic administration of a selective, functionally biased, partial agonist of ADRB1 is effective in reducing neuroinflammation and amyloid beta and tau pathology in the 5XFAD model of AD.