Beta-adrenergic Receptor Agonist Research Articles

Ginsenoside Rg3, enriched in red ginseng extract, improves lipopolysaccharides-induced suppression of brown and beige adipose thermogenesis with mitochondrial activation

Brown adipose tissue (BAT) which is a critical regulator of energy homeostasis, and its activity is inhibited by obesity and low-grade chronic inflammation. Ginsenoside Rg3, the primary constituent of Korean red ginseng (steamed Panax ginseng CA Meyer), has shown therapeutic potential in combating inflammatory and metabolic diseases. However, it remains unclear whether Rg3 can protect against the suppression of browning or activation of BAT induced by inflammation. In this study, we conducted a screening of ginsenoside composition in red ginseng extract (RGE) and explored the anti-adipogenic effects of both RGE and Rg3. We observed that RGE (exist 0.25 mg/mL of Rg3) exhibited significant lipid-lowering effects in adipocytes during adipogenesis. Moreover, treatment with Rg3 (60 μM) led to the inhibition of triglyceride accumulation, subsequently promoting enhanced fatty acid oxidation, as evidenced by the conversion of radiolabeled 3H-fatty acids into 3H-H2O with mitochondrial activation. Rg3 alleviated the attenuation of browning in lipopolysaccharide (LPS)-treated beige adipocytes and primary brown adipocytes by recovered by uncoupling protein 1 (UCP1) and the oxygen consumption rate compared to the LPS-treated group. These protective effects of Rg3 on inflammation-induced inhibition of beige and BAT-derived thermogenesis were confirmed in vivo by treating with CL316,243 (a beta-adrenergic receptor agonist) and LPS to induce browning and inflammation, respectively. Consistent with the in vitro data, treatment with Rg3 (2.5 mg/kg, 8 weeks) effectively reversed the LPS-induced inhibition of brown adipocyte features in C57BL/6 mice. Our findings confirm that Rg3-rich foods are potential browning agents that counteract chronic inflammation and metabolic complications.

Abstract LB123: Conditionally active, therapeutic lymphotoxin beta receptor (LTBR) agonist bispecific antibodies for induction of tertiary lymphoid structures in the treatment of solid tumors

Abstract The presence of tertiary lymphoid structures (TLS) and associated high endothelial venules (HEV) in the tumor microenvironment strongly correlates with improved prognosis and treatment outcomes across a wide range of solid tumors. These clinical observations, now replicated repeatedly, highlight the therapeutic potential of induction of TLS and/or boosting TLS functions, which include acting as a point of entry and education for immune effector cells locally in the tumor. Lymphotoxin beta receptor (LTBR) signalling is essential for lymphoid structure development and the ectopic expression of its ligands, lymphotoxin αβ and LIGHT, is sufficient for TLS formation in vivo but challenging for therapeutic development. Here we present human bispecific antibodies (bsAbs) which conditionally agonize LTBR on co-engagement of fibroblast activation protein (FAP), a tumor microenvironment specific marker expressed by cancer associated fibroblasts. In vitro characterisation of these therapeutic bsAbs is presented, along with in vivo preclinical evaluation of surrogate bsAbs binding to mouse LTBR and mouse FAP in murine tumor models. Results - Therapeutic bsAbs were identified following an extensive screening campaign combining LTBR and FAP binding arms on an engineered human Fc with minimal FcyR binding. In vitro assays using primary cancer associated fibroblasts and LTBR positive cells demonstrated conditionally active therapeutic bsAbs potently activated LTBR in the presence of FAP expressing cells, whereas in the absence of FAP expressing cells no activity was observed. Surrogate bsAbs demonstrated monotherapy activity and led to tumor regressions in combination with anti-PDL1 therapy in an EMT6 mouse model, a syngeneic orthotopic model of breast cancer. Flow cytometry of endpoint tumors showed robust formation of HEVs and increased infiltration of T cells and B cells. In addition, we demonstrate that LTBR agonism, but not PD-L1 inhibition, leads to the formation of TLS structures containing organized lymphocyte aggregates with the appearance of germinal centres and accumulations of T and B cells in a mouse model of lung cancer. In conclusion, conditionally active FAP-LTBR bispecifics activate LTBR in the tumor microenvironment and induce HEV and TLS formation, leading to potent monotherapy activity in vivo, and to tumor regression in combination with PD-L1. These data support the development of FAP-LTBR bispecifics for the treatment of solid tumors as monotherapy and in combination with standard of care. Citation Format: Marta Lewandowska, Fevzi Demircioglu, Lucy Penfold, Rebecca B. Riddle, Sameer Sirohi, Floriane Laurent, Richard Brown, Sonia Bains, Dara Bevan, Emma Stanley, Jeanine Pignatelli, James W. Legg, Ray Jupp. Conditionally active, therapeutic lymphotoxin beta receptor (LTBR) agonist bispecific antibodies for induction of tertiary lymphoid structures in the treatment of solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB123.

Compatibility study of formoterol fumarate-lactose dry powder inhalation formulations: Spray drying, physical mixture and commercial DPIs

Formoterol fumarate (FF) is a long-acting beta-adrenergic receptor agonist that is administered as a dry powder inhaler (DPI) for chronic respiratory conditions. In most DPI formulations, the drug substance and the carrier are the two major components. Lactose is the most common carrier, which can undergo a Maillard reaction in the presence of primary and secondary amines. FF contains a secondary amine. Since the potential for Maillard reaction in DPI formulations has not been investigated yet, this study aims to investigate any potential reaction between lactose and FF. The formulations were prepared using a tubular mixer and spray drying techniques. FF and lactose were blended using a tubular mixer (TM FF-Lac). In the spray drying method, FF and lactose were dissolved in distilled water: MeOH and then spray dried (SD FF-Lac). The samples were stored at both ambient conditions and elevated temperature and humidity (40 °C ± 2, 75% ± 5 RH), and subsequently analyzed to identify any signs of incompatibilities. Differential scanning calorimetry (DSC), field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and liquid chromatography-mass spectrometry (LC-MS-MS) were employed for sample analysis. The in vitro aerosolization performance was evaluated using a next-generation impactor (NGI). The results showed that the colour of the DPI formulations changed from white to brown after storage at elevated temperature and humidity. Although the SD FF-Lac had better aerosolization properties, it was more vulnerable to degradation due to its amorphous nature. The FF-lactose adduct was formed in all samples. Interestingly, adduct compounds were also detectable before the storage for stability and stress tests. Incompatibility was confirmed by HPLC-PDA analysis. The mass spectra revealed the m/z of the adduct compound and confirmed the Millard reaction. Browning occurred in the prepared DPI formulations following storage at elevated temperature and humidity. Moreover, adduct compounds were also found in DPI formulations kept at ambient conditions and commercial DPI. Overall, mass spectroscopy was capable of detecting the incompatibility.

Albuterol as an adjuvant in acute anticholinesterase pesticide poisoning: a randomized, placebo-controlled clinical trial.

Acute anticholinesterase pesticide poisoning is a serious clinical problem, particularly in developing countries. Atropine is the most acceptable treatment for acute anticholinesterase poisoning. However, it only stops fluid production. Albuterol is a beta-2 receptor agonist that can increase fluid removal and speed the return of effective oxygen exchange. This study aims to evaluate the safety and efficacy of nebulized albuterol as an adjuvant therapy in patients with acute anticholinesterase poisoning. This stratified block randomized, single-blinded, placebo-controlled, parallel-group clinical trial was conducted between November 2020 and October 2021. It enrolled 80 patients with acute anticholinesterase pesticide poisoning who were admitted to Tanta University Poison Control Center. Patients were allocated into two groups (40 patients each). The strata were based on the severity of poisoning (moderate and severe). Patients in group I received 10mg of nebulized albuterol. Group II received an equivalent volume of nebulized normal saline. Additionally, standard treatment was provided to both groups. Outcomes included oxygenation, mortality, need for endotracheal intubation and mechanical ventilation, hospital stay duration, time to atropinization, and total doses of atropine and oxime. We found insignificant differences in sociodemographics, exposure characteristics, clinical manifestations, or routine laboratory tests between the studied groups. The median values of oxygen saturation by pulse oximetry were 99% in the albuterol moderate toxicity group and 98% in the control moderate toxicity group. Albuterol significantly improved oxygen saturation in moderate intoxicated patients (P=0.039). Therefore, nebulized albuterol is a safe drug. Moreover, it may improve oxygenation in acute anticholinesterase pesticide poisoning.

Risk assessment in patients with symptomatic and asymptomatic pre-excitation.

Controversy remains as to whether the exercise stress test (EST) is sufficient for risk evaluation in patients with pre-excitation. This study aims to clarify the usefulness of EST in risk stratification in both asymptomatic and symptomatic patients presenting with pre-excitation. This prospective study includes consecutive asymptomatic and symptomatic patients with pre-excitation referred for risk assessment. All participants performed an incremental EST (bicycle) prior to an electrophysiology study (EPS). Primary data from the EST included loss of pre-excitation during exercise, and primary data from the EPS included the measurement of accessory pathway effective refractory period (APERP), shortest pre-excited RR interval (SPERRI), and inducible arrhythmia with the use of a beta-adrenergic receptor agonist if deemed necessary. One hundred and sixty-four patients (59 asymptomatic, 105 symptomatic) completed an EST and EPS. Forty-five patients (27%) demonstrated low-risk findings on EST, of which 19 were asymptomatic and 26 were symptomatic. Six patients with low-risk EST findings had SPERRI/APERP ≤ 250 ms at EPS, and two of them were asymptomatic. The sensitivity, specificity, positive predictive value, negative predictive value (NPV), and accuracy of low-risk EST for excluding patients with SPERRI/APERP ≤ 250 ms were 40, 91, 87, 51, and 60%, respectively. The number of patients with inducible arrhythmia at EPS was similar in the asymptomatic (36, 69%) and symptomatic (73, 61%) groups. Sudden loss of pre-excitation during EST has a low NPV in excluding high-risk APs. The EPS with the use of isoproterenol should be considered to accurately assess the risk of patients with pre-excitation regardless of symptoms (ClinicalTrials.gov Identifier: NCT03301935).

Congenital myasthenic syndromes: beta adrenergic receptor agonist treatment

Acetylcholinesterase inhibitors, such as pyridostigmine, are the standard symptomatic treatment for myasthenia gravis, and so have naturally been applied to the genetic forms of myasthenia, termed congenital myasthenic syndromes (CMS). Although effective for many CMS in others there was no clear response and in some it was positively harmful. Now, with greater understanding of the mutations and molecular mechanisms underlying CMS, treatments can be tailored for the specific syndrome and depending on diseases severity and patient response this can include utilizing different combinations of the drugs. In CMS, over the last 15-20 years b2-adrenergic receptor agonists have moved from occasional use to a mainstream medication. Many cases show life-transforming improvement both when the b2-adrenergic receptor agonists are used alone or in combination. Here we feature how the identification of DOK7-CMS first highlighted the consistent benefit of b2-adrenergic receptor agonists as medication and how it’s application to many different CMS subtypes evolved. The molecular pathogenic mechanisms for many CMS subtypes are now established and this report will also discuss a hypothetical rationale for which forms of CMS are likely to benefit from the b2-adrenergic receptor agonists.

Abstract 1592: The role of beta-adrenergic receptor agonists and inhibitors on the efficacy of Doxorubicin in TNBC cell lines

Abstract Despite the significant advancements in breast cancer (BCa) research and potential therapeutic targets, mortality rates for African American (AA) women remain high compared to other ethnic groups. In the United States, BCa is the second leading cause of cancer-related deaths and is commonly classified by the expression of molecular markers. Treatment and prognosis are determined by the presence and absence of the estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (Her2), and, more recently, the androgen receptor (AR). AA women are more likely to be diagnosed with triple-negative breast cancer (TNBC) and experience a higher risk of BCa mortality than European American women (EA), creating a survival disparity. Biological and non-biological factors are implicated in the difference between EA and AA women contributing to the racial disparities in BCa mortality. Potential implications of this disparity in AA women have been linked to treatment, health care access, socioeconomic status (SES), stress, genetics, and comorbidities. Due to the fraction of available chemotherapeutics for TNBC and the survival disparity in AA women, identifying mechanisms contributing to this aggressive, hard-to-treat BCa is warranted. Studies examining psychological stress and its connection has been shown to contribute to the etiology and aggressiveness of BCa. A direct link has been made in animal models between induced psychological stress and tumor development.Specifically, psychological stress has resulted in higher catecholamine levels and is associated with cancer, resulting in higher incidence, aggression, and mortality. Norepinephrine and epinephrine signal through β-Adrenergic receptors (β-ARs), and these interactions have been shown to decrease apoptosis in prostate cancer cells, increasing malignancy in BCa through the p38 MAPK signaling pathway, and promote BCa stem-like cells. Inhibitors of β-ARs or beta-blockers have been shown to improve the efficacy of chemotherapeutics and decrease the risk of heart failure in cancer patients receiving anthracycline therapy. In this study, we use four TNBC cells (MDA-MB-231, HCC-1143, MDA-MB-468, HCC-1806) that differ in the percentage of West African Ancestry (WAA); we identify differential expression of the ADRB2 protein across all cell lines utilizing flow cytometry. We also show that the non-selective inhibitor of ADRB1 and ADRB2, propranolol, increases the efficacy of Doxorubicin in some cells, which is determined by IC50. We find that the agonist norepinephrine (NE) has no significant impact on cellular growth but impacts the cell's response to Doxorubicin. Here we show that beta-blockers may provide a clinical benefit in combination with Doxorubicin which is dependent upon cell line, and that the β-AR is a promising target for treating TNBC. Citation Format: Alanna Burwell, Portia Andrews, Dayami Lopez, K. Sean Kimbro. The role of beta-adrenergic receptor agonists and inhibitors on the efficacy of Doxorubicin in TNBC cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1592.

Thyroid hormone Beta receptor agonists for treatment of kidney disease: A promising agent?

Chronic kidney disease is a common disorder affecting a significant portion of the adult population with high mortality and morbidity. Obesity and hyperlipidemia are prevalent in chronic kidney disease, and they may trigger fat accumulation in renal parenchyma and eventually fatty kidney. Chronic kidney disease and fatty kidney are also strongly associated with nonalcoholic fatty liver disease. Because they both lead to detrimental effects on organ function, they both need to be treated effectively to improve the outcome. In this narrative review, we have hypothesized that thyroid hormone beta receptor agonists, a novel drug group, may potentially be beneficial in the management of chronic kidney disease due to its promising outcomes among patients with nonalcoholic fatty liver disease, a condition sharing multiple common underlying pathophysiological mechanisms. Thyroid hormone beta receptors are abundantly expressed in liver and kidney tissues, while both nonalcoholic fatty liver disease and chronic kidney disease share various similar pathophysiological mechanisms and triggers. Therefore, thyroid hormone beta receptor agonists may become a promising tool in the management of patients with chronic kidney disease.

Cancer immunotherapies transition endothelial cells into HEVs that generate TCF1+ T lymphocyte niches through a feed-forward loop.

The lack of Tcell infiltrates is a major obstacle to effective immunotherapy in cancer. Conversely, the formation of tumor-associated tertiary-lymphoid-like structures (TA-TLLSs), which are the local site of humoral and cellular immune responses against cancers, is associated with good prognosis, and they have recently been detected in immune checkpoint blockade (ICB)-responding patients. However, how these lymphoid aggregates develop remains poorly understood. By employing single-cell transcriptomics, endothelial fate mapping, and functional multiplex immune profiling, we demonstrate that antiangiogenic immune-modulating therapies evoke transdifferentiation of postcapillary venules into inflamed high-endothelial venules (HEVs) via lymphotoxin/lymphotoxin beta receptor (LT/LTβR) signaling. In turn, tumor HEVs boost intratumoral lymphocyte influx and foster permissive lymphocyte niches for PD1- and PD1+TCF1+ CD8 Tcell progenitors that differentiate into GrzB+PD1+ CD8T effector cells. Tumor-HEVs require continuous CD8 and NK cell-derived signals revealing that tumor HEV maintenance is actively sculpted by the adaptive immune system through a feed-forward loop.

Влияние агониста бета3-адренорецепторов мирабегрона на активность нейтрофилов венозной крови небеременных женщин

The aim of this paper was to determine the effect of the beta3-adrenergic receptor agonist mirabegron on the free-radical activity of venous blood neutrophils in non-pregnant women. Materials and methods. Using the chemiluminescent method (BChL-07 biochemiluminometer), the parameters of the radical response of venous blood neutrophils in 40 non-pregnant women were determined in different phases of the cycle and on the first day after childbirth with additional cell stimulation with latex particles (0.08 μm). When the noise level was automatically subtracted, the following were recorded: maximum intensity of the synthesis of reactive oxygen species, time to reach the maximum of synthesis intensity, and area under the chemiluminogram curve, reflecting the total synthesis of radical particles over the course of 30 min. These parameters were determined before and after an in vitro introduction of the beta -adrenergic receptor agonist mirabegron (10–6 g/l), beta-adrenergic receptor blocker propranolol hydrochloride (10–6 g/l) and agonist against the background of the blocker into the venous blood. Results. Using the Wilcoxon signed-rank test, we showed an increase in the free-radical activity of neutrophils in women in the follicular phase of the menstrual cycle under the influence of mirabegron for all three evaluated parameters. In the luteal phase, mirabegron significantly increased the rate of reaction development and the total synthesis of radical particles, but did not affect the intensity of production of activated oxygen species. However, no effect was observed on the first day after delivery. The possible mechanism of the revealed mirabegron action is discussed in the article. The authors believe that changes in the activity of venous blood neutrophils under the action of mirabegron may be due to changes in the quantity or sensitivity of beta3-adrenergic receptors in different phases of reproduction.

Metabolomics reveals inosine 5′-monophosphate is increased during mice adipocyte browning

Adipocyte browning is one of the potential strategies for the prevention of obesity-related metabolic syndromes, but it is a complex process. Although previous studies make it increasingly clear that several transcription factors and enzymes are essential to induce browning, it is unclear what dynamic and metabolic changes occur in induction of browning. Here, we analyzed the effect of a beta-adrenergic receptor agonist (CL316243, accelerator of browning) on metabolic change in mice adipose tissue and plasma using metabolome analysis and speculated that browning is regulated partly by inosine 5′-monophosphate (IMP) metabolism. To test this hypothesis, we investigated whether Ucp-1, a functional marker of browning, mRNA expression is influenced by IMP metabolism using immortalized adipocytes. Our study showed that mycophenolic acid, an IMP dehydrogenase inhibitor, increases the mRNA expression of Ucp-1 in immortalized adipocytes. Furthermore, we performed a single administration of mycophenolate mofetil, a prodrug of mycophenolic acid, to mice and demonstrated that mycophenolate mofetil induces adipocyte browning and miniaturization of adipocyte size, leading to adipose tissue weight loss. These findings showed that IMP metabolism has a significant effect on adipocyte browning, suggesting that the regulator of IMP metabolism has the potential to prevent obesity.

Aldehyde dehydrogenase 2 and arrhythmogenesis.

Cardiac arrhythmia is a common cardiovascular disease that leads to considerable economic burdens and significant global public health challenges. Despite the remarkable progress made in recent decades, antiarrhythmic therapy remains suboptimal. Aldehyde dehydrogenase 2 (ALDH2), a critical detoxifying enzyme, catalyzes toxic aldehydes and protects individuals from damage caused by oxidative stress. Accumulating evidence has demonstrated that ALDH2 activation has potential antiarrhythmic benefits. The correlation between ALDH2 deficiency and arrhythmogenesis has been widely recognized. In this review, we summarize recent research on the potential role of ALDH2 activation and antiarrhythmic protection, as well as the role played by the ALDH2∗2 polymorphism (rs671) in promoting arrhythmic risk. Additionally, we discuss important new findings illustrating the use of ALDH2 activators, which may prove to be promising antiarrhythmic therapy agents.

Involvement of isoproterenol-induced intracellular Zn2+ dynamics in the basolateral amygdala in conditioned fear memory.

Beta-adrenergic receptors in the basolateral amygdala play an essential role in fear memory, while the physiological role of intracellular Zn2+ remains to be clarified. Intracellular Zn2+ level was decreased 5 min after local injection of 500μM isoproterenol (2μl), a nonselective beta-adrenergic receptor agonist into the basolateral amygdala, suggesting that intracellular Zn2+ dynamic is linked with beta-adrenergic receptor signaling in the basolateral amygdala. When isoproterenol was injected into the basolateral amygdala 20min prior to long-term potentiation (LTP) induction, LTP at perforant pathway-basolateral amygdala was enhanced and conditioned fear memory was also augmented, suggesting that isoproterenol leads to utilization of Zn2+ to consolidate fear memory followed by lowering intracellular Zn2+. We postulated that synaptic Zn2+ dynamics under conditioned fear experience regulates conditioned fear memory in cooperation with beta-adrenergic receptor signaling. When either intracellular Zn2+ chelator (ZnAF-2DA) or extracellular Zn2+ chelator (CaEDTA) was locally injected into the basolateral amygdala in the same manner, LTP was also enhanced. The local injection of ZnAF-2DA augmented fear memory. It is likely that the decrease in availability of intracellular Zn2+ by Zn2+ chelators under fear experience affects the function of Zn2+-required proteins followed by augmentation of fear memory and its related LTP. The present study suggests that beta-adrenergic receptor signaling is linked with intracellular Zn2+ signaling in the basolateral amygdala to consolidate conditioned fear memory. Because intracellular Zn2+ signaling is required for fear memory, the decrease in availability of intracellular Zn2+ may augment fear memory and its related LTP under non-physiological condition.

Select Amino Acid Formulation Influenced Anion Secretion Induced by Th2 Cytokines in Human Bronchial Epithelial Cells (HBECs)

IntroductionAirway surface liquid is tightly regulated by ion channels and/or transporters, and plays a critical role in inflammatory airway diseases, such as ARDS, and in edema fluid clearance during lung injury. Although beta‐adrenergic agonists have been shown to improve alveolar fluid clearance, they have limited effect in late stages of ARDS due to reduced epithelial beta‐adrenergic receptor function or agonist delivery to target regions. CFTR, CLCA2, TMEM16A, and alternative Cl‐/HCO3‐ exchangers (e.g., SLC4 family) are known to be expressed in lung epithelia. Their relative contribution during late stages of lung injury where TH2 cytokines such as IL13 are elevated is not fully understood. The expression pattern of these anion channels are dysregulated in ARDS, and restoration of channel activities could improve fluid accumulation within the lung. In the present study, various anion channel and/or exchanger activities were measured in HBECs exposed to IL13. We have shown that select amino acids influence anion channel activity. Therefore, the effect of a proprietary amino acid formulation on specific anion channels were studied in IL13‐treated HBECs.MethodsTransepithelial short‐circuit current (Isc) was sequentially blocked using inhibitors for ENaC (benzamil), CFTR CFTRInh172), TMEM16A (CaCCInhA01) and Na+/K+/2Cl‐ co‐transporter (bumetanide) in differentiated HBECs mounted in Ussing chambers. Patch‐clamp whole‐cell recordings were performed to determine anion channel activity after IL13 treatment, and channel and/or exchanger expression was analyzed by immunofluorescence.ResultsIL13 increased benzamil‐insensitive (anion) Isc (87.5 ± 4.7 µA.cm‐2 vs 21.9 ± 1.8 µA.cm‐2; P <0.001) and decreased CFTR Isc (1.4 ± 0.52 µA.cm‐2 vs 12.2 ± 0.83 µA.cm‐2; P <0.001) compared to controls. Amino acids reduced benzamil‐insensitive Isc (55.9 ± 4.8 µA.cm‐2; P <0.001) and increased CFTR Isc (6.7 ± 0.75 µA.cm‐2; P <0.001). There was no change in TMEM16A Isc with IL13 treatment. Bumetanide‐sensitive Isc was higher in IL13‐treated cells compared to controls (52.5 ± 3.6 µA.cm‐2 vs 5.5 ± 0.62 µA.cm‐2; P <0.001) suggesting anion secretion via alternate mechanisms, and amino acids decreased bumetanide‐sensitive anion secretion (20.5 ± 2.9 µA.cm‐2; P <0.001). Similarly, whole‐cell patch clamp recordings showed that amino acids restored the CFTR response in IL13‐treated cells. Immunofluorescence imaging demonstrated decreased CFTR expression with IL13, while amino acids recovered CFTR at the apical membrane.ConclusionTreatment with IL13 increased anion secretion while reducing CFTR channel activity in HBECs. Amino acid formulation restored CFTR activity and decreased total anion current in the presence of IL13. These studies suggest anion secretion via alternate anion secretory pathways. Future studies should be directed at understanding the link between CFTR and alternate anion channels in conditions associated with elevated IL13, and how amino acids regulate these channel and/or exchanger functions

Exercise-induced beige adipogenesis of iWAT in Cidea reporter mice

Obesity is caused by an imbalance between energy intake and energy expenditure. Exercise is attracting attention as one of the ways to treat obesity. Exercise induces ‘beige adipogenesis’ in white adipose tissue, increasing total energy expenditure via energy dissipation in the form of heat. Also, beige adipogenesis can be induced by treatment with a beta-adrenergic receptor agonist. We developed a Cidea-dual reporter mouse (Cidea-P2A-Luc2-T2A-tdTomato, Luciferase/tdTomato) model to trace and measure beige adipogenesis in vivo. As a result, both exercise and injection of beta-adrenergic receptor agonist induced beige adipogenesis and was detected through fluorescence and luminescence. We confirmed that exercise and beta-adrenergic receptor agonist induce beige adipogenesis in Cidea-dual reporter mouse, which will be widely used for detecting beige adipogenesis in vivo.

Intracoronary epinephrine and verapamil in the refractory no-reflow phenomenon in patients with acute myocardial infarction

Despite modern advances in performing percutaneous coronary interventions, refractory no-reflow remains a serious problem that worsens in-hospital and long-term prognosis. Low-dose adrenaline may exhibit potent beta-receptor agonist properties that mediate coronary vasodilation.Aim. To evaluate the efficacy and safety of intracoronary administration of epinephrine and verapamil, as well as their combination, compared with standard treatment in patients with ST-segment elevation myocardial infarction (STEMI) and refractory no-reflow during percutaneous coronary interventions.Material and methods. Patients with STEMI and refractory no-reflow will be randomized into 4 groups: standard therapy, intracoronary adrenaline, intracoronary verapamil, intracoronary epinephrine + verapamil. All patients will be assessed for epicardial blood flow using the Thrombolysis in Myocardial Infarction (TIMI) and Myocardial Blush Grade (MBG) scales, peak troponin levels, ST segment changes, echocardiography, magnetic roesnance imaging, and dynamic single photon emission computed tomography.Results. Based on the pharmacodynamic effects of epinephrine and verapamil, their combination is expected to have a more potent vasodilating effect.Conclusion. If the Intracoronary administration of EPInephrine and VERapamil in the refractory no-reflow phenomenon (EPIVER) study will be successful, a novel, more effective method for managing refractory no-reflow phenomenon will appear. This will ensure better preservation of left ventricular systolic function, as well as improve the prognosis and clinical course of the disease.

Role of Tocolytics in Preterm

Tocolytics are the drugs that are used to prolong the time of child birth, for a short period - up to 2 days. If the labour starts too early during pregnancy also termed as preterm birth, which is supposed to be one of the major cause of neonatal mortality and morbidity, all over the world. Administration of tocolytic agents can help in preterm labour to delay the preterm labour by 48 hours. Tocolytics temporarily slow the contractions and hence it is possible to transport the patient to a hospital, where specialized care can be provided to premature baby and also corticosteroids can be administered, which help mature the baby’s lungs. Different class of drugs are studied as tocolytics. Beta-Adrenergic receptor agonists like Terbutaline, Ritodrine, Fenoterol, Metaproterenol, Hexoprenaline, Albuterol, Orciprenaline, Nylidrin. Calcium channel blockers like Nicardipine and Nifedipine. Magnesium Sulfate. Nonsteroidal anti-inflammatories or prostaglandin inhibitors like Indomethacin, and Oxytocin inhibitors like Atosiban were found effective in different studies with some side effects, are discussed here briefly. Some Nitrates, alcohol, Progesterone have also been tried with not much significant effect. Calcium channel blocker specially Nifedipine is most studied and found to be safer and effective for tocolysis. Individual uterine relaxant therapy should take into account individual differences in metabolism and response to drugs.. Short term use of tocolytics has been proven more effective and safer than longer use of it for pre-term labour.

Isoproterenol injected into the basolateral amygdala rescues amyloid β1-42-induced conditioned fear memory deficit via reducing intracellular Zn2+ toxicity

On the basis of amyloid β (Aβ) peptides as triggers in atrophy of structures in the limbic system, here we postulated that Aβ1-42-induced intracellular Zn2+ toxicity in the basolateral amygdala contributes to conditioned fear memory. Aβ1-42 increased intracellular Zn2+ level in the amygdala after local injection of Aβ1-42 into the basolateral amygdala, resulting in conditioned fear memory deficit via attenuated LTP at perforant pathway-basolateral amygdala synapses. Co-injection of isoproterenol, a beta-adrenergic receptor agonist, reduced Aβ1-42-mediated increase in intracellular Zn2+, resulting in rescue of the memory deficit and attenuated LTP. The present study suggests that beta-adrenergic activity induced by isoproterenol in the basolateral amygdala rescues the impairment of conditioned fear memory by Aβ1-42. The rescuing effect may be linked with reducing Aβ1-42-induced intracellular Zn2+ toxicity. Furthermore, Aβ1-42 injection into the basolateral amygdala also attenuated LTP at perforant pathway-dentate granule cell synapses, while co-injection of isoproterenol rescued it, suggesting that Aβ1-42 toxicity in the basolateral amygdala also affects hippocampus-dependent memory. It is likely that beta-adrenergic receptor activation in the basolateral amygdala rescues the limbic system exposed to Aβ1-42 toxicity.

Cell therapy rescues aging-induced beta-1 adrenergic receptor and GRK2 dysfunction in the coronary microcirculation.

Our past study showed that coronary arterioles isolated from adipose-derived stromal vascular fraction (SVF)-treated rats showed amelioration of the age-related decrease in vasodilation to beta-adrenergic receptor (β-AR) agonist and improved β-AR-dependent coronary flow and microvascular function in a model of advanced age. We hypothesized that intravenously (i.v.) injected SVF improves coronary microvascular function in aged rats by re-establishing the equilibrium of the negative regulators of the internal adrenergic signaling cascade, G-protein receptor kinase 2 (GRK2) and G-alpha inhibitory (Gαi) proteins, back to youthful levels. Female Fischer-344 rats aged young (3 months, n = 24), old (24 months, n = 26), and old animals that received 1 × 107 green fluorescent protein (GFP+) SVF cells (O + SVF, n = 11) 4 weeks prior to sacrifice were utilized. Overnight urine was collected prior to sacrifice for catecholamine measurements. Cardiac samples were used for western blotting while coronary arterioles were isolated for pressure myography studies, immunofluorescence staining, and RNA sequencing. Coronary microvascular levels of the β1 adrenergic receptor are decreased with advancing age, but this decreased expression was rescued by SVF treatment. Aging led to a decrease in phosphorylated GRK2 in cardiomyocytes vs. young control with restoration of phosphorylation status by SVF. In vessels, there was no change in genetic transcription (RNAseq) or protein expression (immunofluorescence); however, inhibition of GRK2 (paroxetine) led to improved vasodilation to norepinephrine in the old control (OC) and O + SVF, indicating greater GRK2 functional inhibition of β1-AR in aging. SVF works to improve adrenergic-mediated vasodilation by restoring the β1-AR population and mitigating signal cascade inhibitors to improve vasodilation.

EH Domain-Containing 2 Deficiency Restricts Adipose Tissue Expansion and Impairs Lipolysis in Primary Inguinal Adipocytes

Lipid uptake can be facilitated via caveolae, specific plasma membrane invaginations abundantly expressed in adipocytes. The dynamin-related protein EH domain-containing 2 (EHD2) stabilizes caveolae at the cell surface. Here, we have examined the importance of EHD2 for lipid handling using primary adipocytes isolated from EHD2 knockout (Ehd2−/−) C57BL6/N mice. Following high-fat diet (HFD) feeding, we found a clear impairment of epididymal, but not inguinal, adipose tissue expansion in Ehd2−/− compared with Ehd2+/+ (WT) mice. Cell size distribution analysis revealed that Ehd2−/− mice had a lower proportion of small adipocytes, and an accumulation of medium-sized adipocytes in both epididymal and inguinal adipose tissue. Further, PPARγ activity, FABP4 and caveolin-1 expression were decreased in adipocytes isolated from Ehd2−/− mice. Inguinal adipocytes isolated from Ehd2−/− mice displayed reduced lipolysis in response to beta adrenergic receptor agonist, which was associated with reduced phosphorylation of perilipin-1 and hormone sensitive lipase (HSL). This impairment could not be rescued using a cAMP analog, indicating that impaired lipolysis in Ehd2−/− primary adipocytes likely occurs at the level of, or downstream of, protein kinase A (PKA). Altogether, these findings pinpoint the importance of EHD2 for maintained intracellular lipid metabolism, and emphasize differences in mechanisms regulating lipid handling in various adipose-tissue depots.