bluuebunny/crossref_metadata_2025_split huggingface dataset:
Demand Analysis of the Mu Opioid Receptor Agonist Remifentanil Alone and in Combination with the Cannabinoid Receptor Agonist JWH018
Authors: Maguire David R. , France Charles P. 
Published: June 2021

Opioid abuse is a significant public health problem. With continuing emergence of novel psychoactive substances (e.g., synthetic cannabinoids found in “K2”/”spice” preparations), the co‐administration of opioids and other novel drugs is likely to become more prevalent which might increase the risk for abuse and other adverse effects. This study examined the impact of the synthetic cannabinoid receptor agonist JWH018 on the reinforcing effectiveness of the mu opioid receptor agonist remifentanil in rhesus monkeys (n=5) using economic demand analyses. Lever presses delivered iv infusions of a drug or drug mixture according to a fixed ratio schedule. For each condition, the ratio (10–3200) progressively increased across sessions yielding a demand curve: consumption (infusions obtained) plotted as a function of price (fixed ratio value). When available alone, remifentanil (0.00032 mg/kg/infusion) occasioned the highest unconstrained demand (consumption at the lowest cost) and highest essential value, while JWH018 (0.0032 mg/kg/infusion) alone occasioned lower unconstrained demand and essential value. Unconstrained demand for a mixture of remifentanil and JWH018 was lower than that of remifentanil alone but higher than that of JWH018 alone. Essential value of the mixture was not significantly different from that of remifentanil alone. These data indicate that synthetic cannabinoids such as JWH018 might impact some aspects of opioid self‐administration (i.e., decreased consumption at the lowest price) but do not enhance reinforcing effectiveness as measured by sensitivity of consumption to increasing costs. Opioid/cannabinoid mixtures do not appear to have greater abuse potential compared with an opioid alone.Support or Funding InformationSupported by the NIH (R01DA005018) and the Welch Foundation (AQ‐0039).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of Heat Shock Protein 90 in the Spinal Cord Strongly Promotes Morphine Anti Nociception by Increasing ERK MAPK Activation
Authors: Duron David Isaiah , Lei Wei , Blagg Brian , Streicher John 
Published: June 2021

Hundreds of millions of individuals suffer from acute and chronic pain in the United States alone. The current gold standard of treatment for patients with moderate to severe pain is the use of opioid analgesics. Although the efficacy of these medications is often superior to other options in the clinic, their use is significantly limited due to their side effects and potential for abuse. Recent progress in the research of opioid receptor signaling has shown that manipulation of opioid signaling by biased agonists or other means can improve the therapeutic profile of opioids; however, the field is limited by a lack of understanding of the opioid signaling complex. We have previously shown that inhibition of the chaperone protein Heat shock protein 90 (Hsp90) in the brain strongly blocks opioid anti‐nociception by blocking ERK activation. Here we demonstrate a reversed effect in the spinal cord in that intrathecal (i.t.) injections of the Hsp90 inhibitors 17‐AAG or KU‐32 in CD‐1 mice strongly increase morphine anti‐nociception in paw incision and tail flick pain. Seeking a mechanism for these changes, we found that i.t. Hsp90 inhibition strongly increased ERK activation in the spinal cord in response to opioids, which is opposite of what is observed in the brain, and also implies that Hsp90 acts to repress opioid‐stimulated ERK signaling in the spinal cord. By combining Hsp90 inhibition with the ERK inhibitor U0126 in the spinal cord, we found that ERK inhibition fully reversed the effects of Hsp90 inhibition while having no effect on its own, supporting the hypothesis that increased ERK signaling in spinal cord is responsible for the effects of Hsp90 inhibition. We also performed experiments with systemic and combined i.c.v. and i.t. Hsp90 inhibitor to demonstrate that Hsp90 inhibition in the brain dominates over other circuits when all are inhibited. This data implies that signaling localization within the pain circuit could be key for the targeting of proteins to modify the therapeutic index of opioid analgesics like morphine. These results also suggest that spinal Hsp90 inhibitors could be used as an opioid dose‐reduction strategy which could preserve analgesic efficacy while reducing side effects.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endosomal GPCR signaling: Tyrosine Phosphorylation of a Peptide Linker in NEDD4‐2 Increases Ligase Activity to Promote p38 Proinflammatory Signaling
Authors: Grimsey Neil J. , Rada Cara C. , Narala Rachan , Stephens Bryan S. , Mehta Sohum J. , Lapek John J. , Handel Tracy M. , Zhang Jin J. , Gonzalez David J. , Trejo JoAnn 
Published: June 2021

The regulation of G protein‐coupled receptor (GPCR) signaling is essential for vascular homeostasis and inflammatory responses during disease and injury. MAPK p38 is a key mediator of vascular inflammation, but how GPCRs regulate MAPK p38 signaling is relatively understudied. Post‐translational modifications play a key role in regulating spatiotemporal GPCR signaling and our studies have shown that ubiquitination of GPCRs is not just a cue for trafficking and degradation. Instead, GPCR ubiquitination plays a critical role in endosomal activation of an atypical MAPK p38 signaling pathway. Ubiquitinated receptors recruit the adaptor proteins TAB1 and TAB2 to endosomes and enable auto‐activation of p38, to induce proinflammatory signaling in the vasculature. We have shown that this pathway is conserved for GPCR activation by multiple proinflammatory agents in the vasculature including, thrombin, histamine, ADP and PGE2. Defining how GPCRs activate E3 ubiquitin ligases to initiate endosomal signaling is the focus of our current studies. Using FRET biosensors and phosphoproteomics studies we have discovered that GPCRs activate the E3 ubiquitin ligase NEDD4‐2 by c‐Src dependent tyrosine phosphorylation at a critical tyrosine residue (Tyr485). Tyr485 resides within a predicted auto‐inhibitory linker peptide between the protein binding domains WW II and WW III. Phosphorylation at Tyr485 is required for NEDD4‐2 activation, recruitment to GPCRs and the induction of ubiquitin dependent endosomal p38 signaling. Mutation of Tyr485 blocks thrombin and ADP induced p38 activity and significantly reduces thrombin mediated endothelial permeability. These studies are the first to define how NEDD4‐2 is activated by c‐Src tyrosine phosphorylation, and provides greater clarity into how GPCRs induce proinflammatory p38 signaling from endosomes. Future studies will critical for defining how to selectively block GPCR induced atypical p38 signaling.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of Novel Therapeutic Targets Against Organophosphate‐Induced Axonal Transport Deficits
Authors: Naughton Sean X. 
Published: June 2021

The chemicals known as the organophosphates (OPs) have a wide array of uses in industrial and agricultural settings, most notably as pesticides, but they have also been used on occasion as chemical warfare (nerve) agents. OPs are highly toxic and they have been associated with a variety of acute and long‐term neurologic deficits including impairments in cognition in humans and animal models. The mechanism of the acute toxicity of OPs, cholinesterase inhibition is well documented; however, the neurobiological basis for the long‐term effects of OPs, especially when the exposure levels were below the threshold for acute toxicity is less clear. It is imperative that the mechanism of these deleterious effects of OPs be elucidated so that effective therapeutic strategies can be developed. Previous research in our laboratory conducted both in vivo and in vitro indicated that deficits in axonal transport may represent one mechanism of the long‐term neurologic deficits associated with OPs. In the current series of experiments, we are analyzing post mortem brain tissues from OP‐exposed rats to identify non‐cholinesterase targets that might contribute to the OP‐related deficits in axonal transport. As the axonal transport‐related targets of OPs are identified post mortem, subsequent pharmacological experiments are being conducted in vitro to exploit these targets as a first step toward developing novel therapeutic strategies. In the first series of experiments in tissues collected from rats treated with the OP‐nerve agent, diisopropylfluorophosphate (DFP), 1.0 mg/kg every other day for 30 days, we have identified (using standard immunoblotting techniques) alterations in the phosphorylation of several signaling kinases (e.g., ERK, Akt, and GSKIIβ). We have also identified DFP‐related changes in post‐translational modifications of structural proteins that affect axonal transport through the regulation of microtubule dynamics and stability (e.g., Tau phosphorylation, Tubulin Acetylation). Using a live imaging technique in rat cortical neurons for evaluating fast axonal transport of fluorescent, (APPGFP‐labeled) membrane bound organelles, we have determined that compounds that target signaling kinases (e.g., GSKIIIβ), microtubule stability, and mitochondrial function, can attenuate the DFP‐related impairments in axonal transport. These data not only further elucidate potential non‐cholinesterase targets of OPs, but also provide an initial step toward the design of new therapeutic strategies against OP toxicity.Support or Funding InformationThis work was supported by the Congressionally Directed Medical Research Programs (CDMRP), specifically, the Gulf War Illness Research Program (GWIRP), grant number W81XWH‐12‐1‐0536.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Discovery of MOR Selective, Reversible Opioid Antagonists for Potential Use in Treatment of Drug Dependence
Authors: Jali Abdulmajeed , Obeng Samuel , Zhang Yan , Selley Dana E. 
Published: June 2021

Opioid dependence/addiction is a major public health problem that is associated with multiple health and social costs. Opioids were responsible for the majority of deaths resulting from a drug overdose in 2015 (63% of 52,404). The FDA has approved methadone and buprenorphine for long‐term agonist maintenance therapy. However, a large portion of patients relapse to heroin or illicit use of prescribed opioids. One approach to minimize drug‐induced relapse risk is switching patients to long‐term antagonist therapy after weaning off with an agonist. Naltrexone (NTX) is the only opioid antagonist that is approved for opioid dependence treatment. Nevertheless, NTX is associated with side effects including dysphoria and depression, which contribute to poor patient compliance. Some side effects of NTX could be due in part to its lack of selectivity, as NTX has moderate affinity for the kappa opioid receptors (KOR). Currently, there are no highly‐selective, reversible, non‐peptide μ opioid receptor (MOR) antagonists. Therefore, 18 novel indole‐substituted epoxymorphinan ligands were designed based on the message‐address concept and molecular modeling studies, and synthesized. These novel ligands were hypothesized to bind selectively to the MOR based on interaction with a key aromatic residue in extracellular loop 3 of the MOR (Trp318), such that the added indole substituent in the structure might favor interaction with the indole ring in Trp318 in MOR over KOR and delta opioid receptors (DOR). The purpose of this study was to identify potential MOR‐selective antagonists from this series of compounds using radioligand and [35S]GTPγS binding assays in Chinese Hamster Ovary cells that were stably expressing mouse opioid receptors of interest (MOR, KOR or DOR) and ICR male mouse thalamus. Results showed that all 18 compounds bind to the MOR with high affinity (Ki ≤1 nM) and exhibit a range of efficacies for MOR‐mediated G‐protein activation. Two compounds were discovered to possess most of the desired features. Both compounds have high MOR‐affinity (<1 nM), high MOR selectivity over DOR (>50‐fold) and moderate selectivity over KOR (~8‐fold,) as well as low efficacy to activate the MOR (<20% of the full MOR agonist, [D‐Ala2, N‐MePhe4, Gly‐ol]‐enkephalin (DAMGO)). They also competitively right‐shifted DAMGO‐concentration effect curves for G‐protein activation similarly to naltrexone, indicating they act as competitive MOR antagonists. Our results are encouraging to advance these two compounds for further in‐vitro and in‐vivo pharmacological characterization, and provide important pharmacological information to aid future ligand design.Support or Funding InformationNIH grant R01‐DA024022; Jazan UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
3,3′‐Dichlorobiphenyl (PCB 11) Promotes Dendritic Arborization in Primary Neurons via CREB‐Dependent Mechanisms
Authors: Sethi Sunjay , Keil Kimberly , Wei Feng , Dong Yao , He Guochun , Li Xueshu , Pessah Isaac , Denison Michael , Lehmler Hans‐Joachim , Lein Pamela 
Published: June 2021

PCB 11 is a byproduct of contemporary paint pigment production that has been detected in environmental media around the world, including milk produced for human consumption. We previously reported that PCB 11 enhances dendritic arborization in primary rat neuronal cell cultures. Since increased dendritic arborization, or hyperconnectivity, is a shared characteristic of many neurodevelopmental disorders (NDDs), our findings suggest this chemical poses a risk to the developing brain. The goal of this study was to understand how PCB 11 promotes dendritic growth. We tested canonical mechanisms of PCB developmental neurotoxicity, including signaling mediated by the ryanodine receptor (RyR), aryl hydrocarbon receptor (AhR), thyroid hormone receptor (THR), and cAMP response element binding protein (CREB). [3H]Ryanodine binding assays were used to assess effects of PCB 11 on RyR activity; luciferase‐reporter cell lines, to assess effects on AhR or THR activation. To assess a functional role for these signaling molecules in PCB 11‐mediated dendritic growth, primary neuron‐glia co‐cultures dissociated from neocortices of neonatal Sprague Dawley rats were transfected at 6 days in vitro (DIV) with a MAP2B‐FusRed construct to label the dendritic arbors of individual neurons. At 7 DIV, cultures were pretreated with pharmacological antagonists and then co‐treated for 48 h with PCB 11 or vehicle (0.1% DMSO). Dendritic morphology was quantified in FusRed‐labeled neurons by quantifying the number of dendritic tips and primary dendrites. PCB 11 had no activity at RyR, AhR or THR, and pharmacological antagonism of these receptors had no significant effect on PCB 11‐induced dendritic growth. However, the dendritic promoting effect of PCB 11 was blocked by the selective inhibitor of CREB, 666‐15 (3‐(3‐Aminopropoxy)‐N‐[2‐[[3‐[[(4‐chloro‐2‐hydroxyphenyl)amino]carbonyl]‐2‐naphthalenyl]oxy]ethyl]‐2‐naphthalenecarboxamide), or by co‐transfection with a short hairpin RNA construct specific for CREB. These results indicate that PCB 11 promotes dendritic growth via interaction with CREB or CREB‐dependent mechanism(s), but not directly interfering with known molecular targets of the higher chlorinated PCBs. In light of emerging evidence implicating perturbations of CREB signaling in the etiology of many NDDs, these findings add to the weight of evidence suggesting that PCB 11 poses a threat to the developing human brain.Support or Funding InformationSupported by NIEHS (R01 ES014901 to PJL & INP; T32 ES007059 to SS; P01 ES013661 to HJL) & NICHD (F32 HD088016 to KPK and U54 HD079125).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Retrospective Study of the Relationship Between Testicular Damage and Urinary Creatine Excretion: Possible Markers of Testicular Toxicity Induced by Drugs
Authors: Yono Makoto , Kimura Megumi , Inoue Yumi , Furukawa Kohichiro , Fujiyama Mai , Mugitani Ayumi , Hori Masaharu , Tsuji Shigeki , Tanaka Takanori , Irie Shin , Latifpour Jamshid 
Published: June 2021

ObjectivesCreatine and creatine phosphate act as a buffer system for the regeneration of ATP in tissues with high and fluctuating energy demands. Total creatine content of the rat testis is high compared to all other tissues except skeletal muscle and creatine excretion in rat urine has been shown to increase following testicular damage caused by toxicants. Because the use of biomarkers in toxicology is becoming increasingly important, we investigated whether urinary creatine might be a useful marker for testicular damage in human.MethodsIn a retrospective study, the records of 22 male patients with urological malignancies treated with cisplatin, leuprorelin or goserelin at 4‐week intervals between October 2005 and October 2015 were evaluated. Urinary creatine and serum follicle‐stimulating hormone (FSH), luteinizing hormone (LH) and testosterone levels were assessed in all patients. Single‐voided urine samples were collected to determine urinary creatine‐to‐creatinine ratios. Blood sample collection was done in the morning when serum testosterone levels are around their peak: such morning time sampling for testosterone measurements is recommended in the clinical setting and routinely used in epidemiological studies.ResultsSeven patients with urothelial carcinoma treated with cisplatin based chemotherapy (mean age, 64.1 years) showed no significant changes in urine creatine/creatinine ratio during treatment and had a significant increase in FSH and LH levels and a significant decrease in testosterone levels compared to those at baseline. Similarly, no significant changes in urine creatine/creatinine ratio occurred in patients with prostatic carcinoma treated with leuprorelin (n=8; mean age, 70.0 years) or goserelin (n=7; mean age, 70.4 years), both of whom had castrate levels of testosterone (≤0.5 ng/mL) and suppressed levels of FSH and LH during treatment.ConclusionsOur data demonstrate that testicular damage could not be detected by urinary creatine in patients treated with drugs showing testicular toxicity. Two major testicular functions, i.e., the production of testosterone and formation of haploid germ cells, are generally considered to be regulated by pituitary gonadotrophins, with LH acting on the testosterone‐producing Leydig cells located in the interstitium and FSH affecting Sertoli cells in the seminiferous tubules. Thus, our hormonal data may indicate the presence of cisplatin‐induced Sertoli cell dysfunction and inhibition of Leydig cell function by LH‐RH analogues.Support or Funding InformationNoneThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
SNPs Outside Response Elements Impact Aryl Hydrocarbon Receptor (AHR) Binding and Gene Regulation: Genome‐wide SNP‐dependent Transcriptional Regulation
Authors: Neavin Drew , Liu Duan , Li Hu , Wang Liewei , Weinshilboum Richard 
Published: June 2021

BACKGROUNDMost functionally‐significant single nucleotide polymorphisms (SNPs) are outside of open reading frames. Mechanisms by which these SNPs contribute to gene regulation are poorly understood. Aryl hydrocarbon receptor (AHR) is a nuclear receptor that regulates drug metabolism, xenobiotic response and immune response. Our previous data indicate that SNPs outside response elements can impact transcription factor binding and gene regulation. We hypothesized that SNPs near but outside AHR response elements (AhREs) might impact AHR binding and gene regulation.METHODSWe have collected dense SNP‐genotypes across 300 lymphoblastoid cell lines (LCLs) – a powerful tool for generating and testing pharmacogenomic hypotheses. Eight LCLs were used to generate hypotheses about AHR SNP‐dependent transcription utilizing RNA‐ and ChIP‐seq, which were tested in additional LCLs using qRT‐PCR and ChIP after AHR agonist or antagonist treatment.RESULTS69 genes were highly significantly differentially expressed after agonist or antagonist treatment and 80% of those genes had an AhRE within 50kb. Five SNP‐gene pairs have been functionally validated in additional LCLs (Table 1).CONCLUSIONSNPs that are outside of AhREs can alter AHR binding and gene transcription. This is a general phenomenon that may have large‐scale influence on gene transcriptional regulation and pharmacogenomics.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulation of KRas‐Mediated Signaling by G‐Protein Coupled Receptors in Pancreatic Cancer
Authors: Feigin Michael E. 
Published: June 2021

Pancreatic ductal adenocarcinoma is a highly lethal malignancy with few effective treatment options. We used computational approaches to identify new drug targets within this heterogeneous patient population, focusing on the G‐protein coupled receptors (GPCRs). GPCRs transduce extracellular signals from a variety of ligands through the plasma membrane, resulting in the modulation of intracellular signaling pathways and regulation of many physiological processes. This wide array of functions has resulted in the extensive utilization of GPCR targeted therapeutics, accounting for 30–50% of all currently used drugs. However, drugs targeting GPCRs are rarely utilized in cancer treatment, despite evidence that GPCRs mediate many aspects of tumorigenesis. Genomic analyses have uncovered GPCR mutations, copy number alterations and gene expression and methylation changes in a wide variety of cancers. Determining the biological implication of these genomic alterations will allow utilization of GPCR targeted therapeutics in those patients with GPCR‐driven tumors. Our preliminary evidence has uncovered the orphan GPCR GPRC5A as a critical regulator of the progression from early stage PanIN lesions to PDA.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.