bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of omega‐3 and omega‐6 fatty acids on nitric oxide and IL‐6 production by RAW 264.7 macrophages
Authors: Danchine Veronika Denise , Caldari Cristina 
Published: June 2021

Chronic low‐grade inflammation, or meta‐inflammation, which may occur in obese subjects, is related to the infiltration of macrophages into adipose tissue (AT) as adipocyte number and size increase. The increased count in fat cells is correlated with an increase in adipose tissue macrophages (ATMs). These ATMs can secrete pro‐inflammatory cytokines which can have effects on the surrounding adipocytes. Omega‐3 fatty acids (FAs) reduce inflammation through direct competition with omega‐6 FAs, the precursors of the inflammatory eicosanoids, as well as through changes in gene expression of inflammatory molecules. Less is known about the effects of omega FAs on pro‐inflammatory cytokine production by immune cells, such as macrophages. The objective of this research was to determine the effects of omega‐3 and omega‐6 FAs on the production of nitric oxide (NO), a signaling molecule secreted by activated macrophages which is involved in mediating inflammation and production of the pro‐inflammatory cytokine interleukin‐6 (IL‐6), in lipopolysaccharide (LPS)‐activated macrophages. RAW 264.7 (murine macrophages) were incubated with 100 μM of sodium salts of linoleic acid (LA; C18:2, omega‐6), arachidonic acid (ARA; C20:4, omega‐6), or eicosapentaenoic acid (EPA; C20:5, omega‐3) for 24 h. After pre‐incubation with FAs macrophages were stimulated with 0.01 μg/ml of LPS for 6 h. Conditioned media was collected for NO and IL‐6 quantification via Griess assay and ELISA, respectively. FA treatments were cytoprotective, resulting in higher percentages of live cells than both control and LPS‐treated cells (Trypan blue exclusion method; p<0.0001, two‐way ANOVA). LPS‐stimulated macrophages pre‐incubated with LA, ARA, or EPA produced less NO than LPS‐stimulated macrophages without FA pre‐incubation (p=0.0001, two‐way ANOVA). Pre‐incubation of macrophages with LA, ARA, or EPA decreased IL‐6 production in comparison to LPS‐stimulated macrophages, with omega‐6 FAs having a more pronounced effect than omega‐3 FAs (p<0.0001, two‐way ANOVA). In conclusion, both omega‐6 and ‐3 FAs decreased activation of murine macrophages and decreased production of the inflammatory molecules NO and IL‐6, with omega‐6 FAs having a more marked effect than omega‐3 FAs. The stronger anti‐inflammatory effect of omega‐6 FAs on murine macrophages, compared to omega‐3 FAs, warrants further investigation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelial Extracellular Vesicle Production and Cargo Property are Altered after Inflammatory Injury
Authors: Chatterjee Victor , Yang Xiaoyuan , Yuan Sarah Y. 
Published: June 2021

Extracellular vesicles (EVs) play an important role in cell‐cell communication and signal transduction by transferring bioactive cargo contents between cells or tissues. During inflammation, the vascular endothelium is activated and releases vasoactive EVs that alter vascular homeostasis via paracrine or autocrine pathways. In this study, we examined the molecular property and mechanisms of production of EVs derived from endothelial cells (EC) during septic or sterile (thermal injury) inflammation, and during stimulation by inflammatory agonists, including tumor necrosis alpha (TNFa), interleukins (ILs), lipopolysaccharide (LPS) and thrombin. EVs isolated from plasma or cell culture conditioned media were analyzed by flow cytometry using cell specific markers. Size‐based gating of microvesicles (MV) was performed using a mixture of standardized beads of different sizes and absolute MV numbers calculated based on reference beads of known concentration. Immuno‐capture of smaller exosomes was performed with anti‐CD9 coated beads, probed for proteins of interest and visualized by flow cytometry. The results revealed that mice subjected to cecal ligation and puncture‐induced septic injury or thermal injury had increased CD144+ EVs but decreased endothelial nitric oxide synthase (eNOS)+ EVs in plasma as compared to sham‐treated mice. Likewise, endothelial cells stimulated by TNFa, IL‐1b or thrombin release EVs containing a higher level of caveolin‐1 (Cav‐1) and lower level of eNOS compared to non‐stimulated cells. Pre‐treatment with the ROCK inhibitor Y27632 prevented this response. Furthermore, EV‐EC interactions were observed under confocal microscopy. In endothelial cells, internalization of inflammatory EVs induced eNOS translocation from the cell membrane to cytosol. These findings suggest that inflammation alters endothelial production of EVs and their cargo property and function.Support or Funding InformationThis work was supported by NIH grants HL070752, HL126646 and GM097270This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Keystone Species in the Pathogenic Process of NAFLD
Authors: Fang Sa , Wu Dingfeng , Yang Zihuan , Jiao Na , Lin Yunzheng , Li Xiaoyi , Xiao Jing , Yuan Meifei , Zhu Ruixin , Zhu Lixin 
Published: June 2021

BackgroundThe role of gut microbiome in the pathogenic process of NAFLD has been increasingly recognized. However, the keystone species in the gut of NAFLD, the critical factor(s) in maintaining the stability of the entire gut microbiome, has not been reported.MethodSparCC method was used to construct the microbial interaction ecosystem of human gut microbiome in healthy condition. PageRank algorithm was used to evaluate the importance of each bacterial species in healthy gut micro‐ecology system. Subsequently, the keystone species required for maintaining the stability of gut micro‐ecological system was identified on the basis of network stability assessment. Finally, the keystone species specific for NAFLD was determined by the quantitative analysis of gut microbiome between NAFLD patients and healthy people. And the relevant OTUs were annotated at species‐level based on phylogenetic trees. Furthermore, the same keystone species identification analysis was also conducted in NAFLD rats induced by high‐fat diet.ResultsHere we reconstructed the microbial interaction ecosystem of healthy human gut microbiome, consisting of 61 bacterial species and 90 pairwise interactions. Wherein, Bacteroides ovatus, Faecalibacterium prausnitzii, Barnesiella intestinihominis, Odoribacter splanchnicus, Roseburia faecis, Eubacterium ventriosum, Clostridium leptum were the seven keystone species that maintained the stability of the gut micro‐ecological system. The results of quantitative analysis indicated that the abundance of B. intestinihominis, E. ventriosum and C. leptum decreased significantly in the gut of NAFLD patients, and they were identified as the keystone species specific for NAFLD. These species could participate in the initiation and development of NAFLD by regulating the metabolic process of bile acid signaling and the activity of inflammasome. In addition, similar results were observed in the gut microbiome of NAFLD rats.ConclusionThis research identifies the keystone species specific for NAFLD, and highlights a novel target for the precise prevention and treatment of NAFLD based on gut microbiome. Importantly, our strategy for the identification of the keystone species can be extended to other ecological systems.Support or Funding InformationThis work was supported by National Natural Science Foundation of China 81770571 (to LZ), 81774152(to RZ), 41530105 (to RZ).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Rapastinel as a non‐opioid treatment for opioid dependence
Authors: Ferrante Julia Renee , Lamichhane Nidesh , Kuhn Cynthia , Armstrong Christopher , Walker David , Patkar Ashwin 
Published: June 2021

An increasing number of opioid overdose deaths have resulted from the rising availability of both prescription and nonprescription opioids. Pharmacotherapies, such as buprenorphine and methadone, are used to treat those dependent on opioids. However, these commonly used pharmacotherapies are opioid partial agonists and agonists, so the patient remains in an opioid‐dependent state throughout treatment. Treatment requires long‐term tapering with buprenorphine or methadone, during which withdrawal symptoms can occur. These limitations show a clear need for a non‐opioid pharmacotherapy. Drugs that target the NMDA receptor have been proposed as alternatives to opioid‐targeted pharmacotherapies. The NMDA receptor is involved in the maintenance of opioid dependence and its blockade may reverse the neuroadaptive changes induced by opioid dependence. Regulation of the NMDA receptor could accelerate treatment of opioid dependency (Glass, 2011). Ketamine, an NMDA antagonist, has potential as a treatment, but has associated side effects and potential for abuse. Rapastinel is a novel drug marketed as an antidepressant, and it acts as a partial agonist of the NMDA receptor complex. The negative side effects reported with ketamine have not been reported with rapastinel (Moskal et al., 2016). Since rapastinel acts as a partial agonist and has no reported side effects, it may be a more tolerable treatment option for those dependent on opioids. Male and female adolescent (between 28 and 30 days old) Sprague‐Dawley rats were injected with morphine in increasing doses twice a day for five days (5 mg/kg/day–25 mg/kg/day) to induce opioid dependency. On day six, rats were injected with naloxone (1 mg/kg) and withdrawal signs were quantified. Rats were then given either ketamine injections (1 mg/kg) twice daily (n=12), rapastinel injections (5 mg/kg) every other day (n=14), or saline injections (n=24). On day nine, rats were again administered naloxone (1 mg/kg), and withdrawal signs were quantified to measure efficacy of ketamine and rapastinel. Rapastinel treated rats exhibited significantly fewer withdrawal signs than those treated with ketamine, which did not differ from controls. These results show that rapastinel significantly enhances recovery from opioid dependence.Support or Funding InformationFunding provided by ASPET, Duke University SURPH program, and Duke University Department of Pharmacology and Cancer BiologyThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A novel conformation‐sensitive antibody‐based strategy to explore PKC signaling induced by opiate activation of μ‐opioid receptors
Authors: Duarte Mariana Lemos , Devi Lakshmi 
Published: June 2021

Opiates such as morphine and fentanyl are the most widely used analgesics for the treatment of pain; however, prolonged treatment with opiates decreases the responsiveness of opioid receptors, leading to the development of tolerance and opiate addiction. Recent alarming reports of the widespread misuse of fentanyl leading to the ‘Opioid Epidemic” have begun to refocus scientific investigations into the mechanisms of action of opioid receptors. Despite intense research efforts, the molecular mechanisms underlying opioid addiction remain unknown contributing to a public health challenge. Activation of opioid receptors leads to the initiation of a number of signaling cascades, resulting in enhanced protein phosphorylation. Among the various kinases, protein kinase C (PKC) has been reported to play a crucial role in the desensitization of opioid receptors. To date, the majority of studies with PKC have used genetic and/or pharmacological manipulation of PKC to explore the dynamics of PKC activation. In order to investigate the activation of endogenous PKC in native tissue, we developed conformational‐sensitive antibodies that are able to detect time‐ and ligand‐mediated activation of native PKC. We also generated antibodies to several substrates of PKC including pPP2A (regulatory subunit phosphoS90 PPP2R5D), phosphoSer143 MARCKS, and phosphoS375/T376 MOR. We used a high‐throughput microscopy approach to explore the temporal dynamics of opiate‐mediated PKC signaling and track the temporal dynamics of PKC activation in cells treated with PMA, morphine or fentanyl. Interestingly, in contrast to the PKC activation induced by morphine that is rapid but transient the activation by fentanyl is slow and sustained for a longer period of time. We are able to detect rapid activation of PKC in the striatum of animals acutely treated with morphine suggesting that the conformation‐sensitive activation, state sensitive anti‐PKC antibodies serve as hitherto unavailable, critical tools to investigate the role of PKC in opioid receptor signaling in particular and PKC‐mediated signaling in general.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Icariside II, a phosphodiesterase 5 inhibitor, attenuates brain injury in focal cerebral ischemia/reperfusion rats: Involvement of inhibition of GSK‐3β‐mediated activation of autophagy
Authors: Gao Jianmei , Li Fei , Long Long , Xu Fan , Feng Linying , Liu Yuangui , Shi Jingshan , Gong Qihai 
Published: June 2021

Cerebral ischemia/reperfusion (I/R) is always exacerbated neuronal damage when timely restoring cerebral blood flow after ischemia stroke. Cerebral I/R results in excessive autophagy and leads to neuronal demise. The present study was designed to investigate the role of autophagy during ICS II‐mediated neuroprotection on cerebral I/R in vivo and in vitro. Here, we found that ICS II not only protected against middle cerebral artery occlusion‐induced injury in rats, but also attenuated primary cortical neurons injury upon oxygen‐glucose deprivation. Moreover, autophagy was activated as evidenced by triggered autophagic influx and increased ATG expressions after stimulation of cerebral I/R. However, ICS II signifcantly inhibited excessive autophagy as evidenced by repressing autophagic flux, combined with decreasing ATG expressions including LC3 II, Beclin 1, ATG5 and ATG7. Interestingly, ICS II, a PDE5 inhibitor, not only restored cGMP/PKG pathway, but also inactivated GSK‐3β through mediating phosphorylation at tyr216 and ser9. Most intrugingly, siGSK‐3β obviously promoted the beneficial effect of ICS II, which further confirmed that the neuroprotective mechanism of ICS II is, at least in partly, through inactivating GSK‐3β. Furthermore, we reveled that the cGMP/PKG pathway acts as the upstream of GSK‐3β inactivation in the inhibitory effect of ICS II on cerebral I/R‐induced autophagic neuronal death, as evidenced by GSK‐3β inhibitor or PKG inhibitor in vitro. Most importantly, the findings demonstrated that ICS II protected against cerebral I/R‐induced injury via mediating PKG/GSK‐3β/autophagy axis. This study will raise the possibility that ICS II may be exploited into a potent PDE 5 inhibitor against ischemia stroke.Support or Funding InformationThis work was supported by Natural Science Foundation of China Natural Science Foundation of China (Grant No. 81560666), Program for excellent young talents of Zunyi Medical University (Grant No. 15zy‐002), Science and Technology Innovation Talent Team of Guizhou Province (Grant No.20154023), The hundred level of high‐level innovative talents in Guizhou Province (Grant No. QKHRCPT 20165684), and Program for Changjiang Scholars and Innovative Research Team in University, China (Grant No. IRT_17R113).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modulatory Effect of a Standardised Andrographis Paniculata Aqueous Extract on Urine Metabolomics in a Rat Model of Neuroinflammation
Authors: Sani Dahiru , Kirby Brian P , Basri Hamidon , Stanslas Johnson 
Published: June 2021

In this study, the effect of pre‐treatment with standardised andrographis paniculata aqueous extract (APAE) on rats urine metabolomic changes following Lipopolysaccharide (LPS)‐induced neuroinflammation was evaluated based on proton‐nuclear magnetic resonance (1H‐NMR) spectroscopy. Both principal component analysis and orthogonal Partial Least Square‐Discriminant Analysis (OPLS‐DA) revealed that LPS (1 mg/kg) induces metabolic alterations. LPS group showed increased urinary levels of oxoglutarate, creatinine, citrate, allantoin, lactate and hippurate. However, rats pre‐treatment with APAE for 7 days prior to LPS administration prevented the upregulation of these molecules similar to the control group to confer neuroprotection. Our findings imply that metabolomic procedures based on NMR could be applicable to elucidate neuro‐inflammation via the biomarkers exploration. In addition, we proposed a mechanistic metabolic pathway for APAE suppression of neuroinflammation.Support or Funding InformationThis study was supported by the Malaysia Ministry of Agriculture (NRGS grant, NH612D009) and by Government of Sokoto State, NigeriaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Specific TBC Domain‐Containing Proteins Control the ER‐Golgi‐Plasma Membrane Trafficking of GPCRs
Authors: Wei Zhe , Huang Wei , Wu Guangyu 
Published: June 2021

G protein‐coupled receptors (GPCRs) constitute the largest superfamily of cell‐surface signaling proteins. However, the molecular mechanisms underlying their cell surface delivery after synthesis remain poorly understood. Here we screen the TBC domain‐containing proteins, putative Rab GTPase‐activating proteins (GAPs), in the intracellular trafficking of GPCRs and identify several TBC proteins that activity‐dependently regulate the anterograde transport, en route from the endoplasmic reticulum to the Golgi or from the Golgi to the cell surface, of several prototypic GPCR members without affecting other plasma membrane proteins. We also show that TBC1D6 functions as a GAP for Rab26, physically associates with Rab26, and attenuates Rab26 interaction with GPCRs. Furthermore, both overexpression and depletion of TBC1D6 inhibit the post‐Golgi traffic of GPCRs. These data demonstrate for the first time the important roles of the TBC proteins in forward trafficking of nascent GPCRs and reveal novel regulatory mechanisms of GPCR targeting to the functional destination.Support or Funding InformationNIH R01GM118915This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Therapeutic Assessment and Neuroprotective Effects of a Novel G Protein‐Coupled Estrogen Receptor (GPR30/GPER) Agonist
Authors: Sondergard Chelsea , Gunn Joseph , Pemberton Kyle , Arnatt Christopher K 
Published: June 2021

The G protein‐coupled estrogen receptor (GPER) has been implicated in the neuroprotective effects of 17β‐estradiol and selective GPER agonist, G‐1. Currently, G‐1 remains the standard for GPER agonists, necessitating the design and identification of novel candidates. Previously, our group synthesized and biologically evaluated a series of GPER antagonists. Modification of an aromatic moiety on the scaffold altered the pharmacological profile from antagonists to agonists. Using a fluorescence‐based binding assay for the classical estrogen receptors, ERa and ERb, we discovered off‐target binding. Due to the similarity in functionalization to estradiol, we hypothesized that the hydrogen bonding abilities of the initial candidate were responsible for the observed off‐target binding. To mitigate off‐target binding, modifications were made to reduce the hydrogen bonding ability. For each resulting candidate, the EC50 value was determined through in vitro biochemical screening. From these studies, we identified a lead compound that exhibited an EC50 value below that of our experimentally determined EC50 value of G‐1. Using E18 rat embryonic hippocampal neurons, the lead compound was shown to increase synaptogenesis. These results suggest that the identified lead compound for GPER may be a potential candidate for neurodegenerative diseases.Support or Funding InformationSaint Louis UniversityThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Marine Pharmacology and Pharmaceuticals Pipeline in 2018
Authors: Abadines Ivan B , Le Kelly , Newman David J , Glaser Keith B , Mayer Alejandro M 
Published: June 2021

The ocean's organisms continue to deliver a chemical biodiversity that contributes to both the global preclinical and clinical pharmaceutical pipelines. Thus, in November 2018, the clinical marine pharmaceutical pipeline consisted of seven marine‐derived drugs approved by the United States Food and Drug Administration (FDA): for cancer, cytarabine (Cytosar‐U®, Depocyt®, FDA‐approved 1969); for Herpes Simplex Virus, vidarabine (Vira‐A®, FDA‐approved 1976); for pain, ziconotide (Prialt®, FDA‐approved 2004); for hypertriglyceridemia, omega‐3‐acid ethyl esters (Lovaza®, FDA‐approved 2004); for cancer, eribulin mesylate (Halaven®, FDA‐approved 2010), brentuximab vedotin (Adcetris®, FDA‐approved 2011), and trabectedin (Yondelis®, FDA‐approved 2015). The website www.clinicaltrials.gov was searched to determine marine‐derived pharmaceuticals that were in Phase I, Phase II and Phase III of clinical development in November 2018. Confirmation of our findings in the www.clinicaltrials.gov website was conducted by visiting the websites of all pharmaceutical companies supporting the development of compounds currently in Phase I, Phase II and Phase III of clinical development. Our clinical research determined that there were 25 marine‐derived compounds in active clinical trials in November 2018: 6 marine‐derived compounds were in Phase III, 10 compounds in Phase II, and at least 9 compounds in Phase I, many of them auristatin‐containing antibody drug conjugates or ADCs. Regular updates on the clinical marine pharmaceutical pipeline will be posted on the dedicated website: http://marinepharmacology.midwestern.edu/clinical_pipeline.html. Furthermore, it is important to note that the global preclinical marine pharmacology pipeline continues to generate considerable data on multiple pharmacological classes, as highlighted in a recent review: A. M. S. Mayer, A. D. Rodríguez, O. Taglialatela‐Scafati and N. Fusetani. Marine Pharmacology in 2012–2013: Marine Compounds with Antibacterial, Antidiabetic, Antifungal, Anti‐Inflammatory, Antiprotozoal, Antituberculosis, and Antiviral Activities; Affecting the Immune and Nervous Systems, and other Miscellaneous Mechanisms of Action. Marine Drugs 15 (9): 273, 2017. We thus conclude that in November 2018, both the marine pharmacology preclinical and clinical pharmaceutical pipelines continued to remain very active.Support or Funding InformationSupport by the College of Pharmacy, Vicki Sears, Pharmacology Department, College of Graduate Studies and Elizabeth Tallman, Information Technology Services, Midwestern University are gratefully acknowledged.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.