bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mu Opioid Receptor Agonists with Dopamine D3 Receptor Antagonism: Toward Developing Analgesics with Less Abuse Potential
Authors: Shah Nilay M. , Schmid Cullen L. , Merritt Benjamin , Kennedy Nicole M. , Ross Nicolette C. , Lovell Kimberly M. , Bannister Thomas D. , Bohn Laura M. 
Published: June 2021

Opioid analgesics such as morphine are widely used in clinical practice for the management of moderate to severe pain. While they are the gold standard for painkillers, they are also highly addictive: approximately 10% of patients prescribed opioids for pain‐relief develop an opioid‐use disorder. Abuse of prescription opioids and heroin has become an epidemic in the United States with opioid overdose deaths quadrupling since 1999. Opioids induce their physiological effects through the activation of the mu opioid receptor (MOR). Interestingly, co‐administration of dopamine D3 (DRD3) receptor antagonists with opioids attenuates the self‐administration and rewarding effects of the drugs, but has no effect on their ability to relieve pain. This suggests that the development of compounds that both activate MOR and antagonize DRD3 may be a means to maintain the beneficial effects of opioids, while reducing their abuse potential. We have recently developed a library of small molecules with a novel scaffold that are agonists at MOR. Herein we screened our MOR agonists for antagonism at DRD3 in a DRD3/βarrestin enzyme fragment complementation assay. We identified multiple compounds that antagonize DRD3 that are as potent as Sulpiride, a known DRD3 antagonist. Further characterization of these MOR agonists with polypharmacology at DRD3 may allow for the development of opioid analgesics with reduced abuse liabilities.Support or Funding InformationFunded by NIDA grant R01 DA033073 to LMB Fellowship to NMS: NSF award 1359369. and TDB, and NSF Undergraduate ResearchThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Autophagy During Myocardial Ischemia/Reperfusion Injury
Authors: Ibe Al C. , McIntyre Anahi , Kim Alissa , Kim Hannah , Castellano Andrew , Young Lindon , Barsotti Robert , Chen Qian 
Published: June 2021

Autophagy is a housekeeping process to degrade protein aggregates, damaged cytoplasmic constituents, and provides adaptation when cells go through a nutrient stress. However, a debate persists on whether autophagy is beneficial to attenuate or promote cell damage under an ischemia/reperfusion (I/R) insult to the heart. This study tested the effects of an autophagy enhancer (e.g. rapamycin and trehalose) on heart function and infarct size after global ischemia (30 min)/reperfusion (45 min) when given prior to ischemia (pretreatment) or at the beginning of reperfusion (posttreatment). We found that rapamycin pretreatment (25 nM, n=5) significantly restored left ventricular developed pressure (LVDP) to 64±5 % of initial baseline compared to control I/R hearts (n=8) which only recovered to 31±10% of initial baseline (p<0.01). Likewise, trehalose pretreatment (5 mM, n=7) and posttreatment (5 mM, n=6) significantly restored final LVDP to 62±5% and 65±4% of initial baseline, respectively compared to control I/R hearts (both p<0.01). Moreover, infarction percentage after I/R is significantly reduced in rapamycin post‐treated hearts (23±4%, p<0.05) and trehalose treated hearts (20±4% for pretreatment and 15±3% for posttreatment, both p<0.01) when compared to control I/R hearts (41±4%). The preliminary data suggest that autophagy enhancement before ischemia or at reperfusion may be beneficial for reducing I/R injuryThis study was supported by the Center for Chronic Disorders of Aging, the Division of Research and the Department of Bio‐Medical Sciences at Philadelphia College of Osteopathic Medicine.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of Hypoxia and Inflammation on Claudin‐1 in CaCo‐2 Cells as a Model Intestinal Epithelium.
Authors: Largoza Gabrielle E. , Cole Chelsea E. , Alana Nicholas B. , Carr Nicholas R. , King Jonathan M. 
Published: June 2021

BackgroundThe use of hydrocortisone in association with indomethacin is a known risk factor for spontaneous intestinal perforation in extremely low birthweight infants. Hypoxia, inflammation and non‐steroidal anti‐inflammatory drugs independently alter apical junctional complexes of epithelial cells. We are interested in understanding the combined effects of hypoxia, inflammation and non‐steroidal anti‐inflammatory drugs on the apical junctional complex and barrier integrity in a gastrointestinal epithelial cell model.MethodsThrough the use of CaCo‐2 BBe monolayers, an established in vitro model of the intestinal epithelium, we analyzed the effects of the pro‐inflammatory agents (TNF and IFN‐gamma) and anti‐inflammatory agents (indomethacin and hydrocortisone) on junctional protein expression and localization, cell proliferation using Edu labeling, cellular ATP concentration and barrier function.ResultsProtein expression analyzed by Western blotting techniques demonstrated an alteration of claudin‐1, a tight junction sealing protein. The soluble and insoluble fractions of the cells were analyzed in order to determine the location of these proteins. We determined inflammatory conditions produced a higher concentration in the soluble fraction indicating impaired function. Confocal analysis after 24‐hour exposure to hypoxia showed that the combined use of hypoxia and inflammatory agents elicited no significant change in cell proliferation levels. Cellular ATP levels were analyzed to corroborate proliferation studies. Confocal imaging under the same experimental conditions was also conducted in order to probe for claudin‐1 localization within the cells.ConclusionsThere was no observed significant difference in the cell proliferation rate or marked changes in ATP levels in our model. Therefore, we hypothesize that disturbance in barrier function may be related to specific junctional mechanisms. We found claudin‐1 protein expression and localization to be sensitive to both hypoxia and inflammatory cytokines contributing to impaired barrier integrity. These cellular studies suggest claudin‐1 stability may be a potential target in the progression of spontaneous intestinal perforation.Disclosure: The view(s) expressed herein are those of the author(s) and do not reflect the official policy or position of Brooke Army Medical Center, the U.S. Army Medical Department, the U.S. Army Office of the Surgeon General, the Department of the Air Force and Army, Department of Defense or the U.S. Government.Support or Funding InformationSan Antonio Medical Foundation Grant, Trinity University, US Air Force 59th Medical Wing Clinical Research WingThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Organophosphate Pesticide Effects on Neuronal Cytoarchitecture and Morphological Abnormalities in 72 hour Zebrafish Embryos
Authors: Watanabe Taylor H. , Fradinger Erica A. 
Published: June 2021

Organophosphate pesticides (OP) are a class of acetylcholinesterase (AChE) inhibitors, which have effects on the central and peripheral nervous systems. OP's inhibit AChE through phosphorylation of the active site serine, which leads to a toxic and potentially lethal buildup of the neurotransmitter acetylcholine (ACh) causing excess stimulation of cholinergic neurons. OP's are widely used to suppress the activity and growth of insect pest populations in both agriculture and household settings. The objective of this study was to examine both organismal and cellular morphological changes caused by OP exposure. Specifically, zebrafish (Danio Rerio) embryos were exposed to sub‐lethal concentrations of Chlorpyrifos (CPF) and its bioactive form Chlorpyrifos‐oxon (CPO), and compared to vehicle controls (MeOH). Gross morphological changes were observed at 72 hours via light microscopy and morphological changes to cholinergic neurons were observed via confocal microscopy of wholemount immuno‐labeled embryos using anti‐ChAT. Light microscopy observations of the embryos showed varying degrees of cardiac edema in both CPF and CPO exposed embryos. Additionally, mild kyphosis was observed in CPF and strong kyphosis in CPO exposed embryos. Confocal observations of the trunk spinal region showed embryos exposed to CPF lost dorsal neural tube cell staining and dark pockets, possibly indicating cell death, were observed along the ventral neural tube. For CPO exposed embryos, cells bodies in the neural tube were disorganized. Additionally, axon projections across the neural tube showed differences in branching and spacing in both CPF and CPO exposed embryos. Together our results showed that OP exposure causes changes in neuronal cytoarchitecture and embryo morphology. Understanding the developmental effects of these pesticides is particularly important since developmental changes have significant effects on adult systems and children represent a vulnerable population.Support or Funding InformationDepartment of Defense, ARO 66377‐RT‐REP; Keck Foundation Undergraduate Fellowship to T.H.W.; James Irvine Foundation Chair in Biological Sciences to E.A.F.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cdc48a AAA‐ATPase and its association with ubiquitin‐like SAMP1 and DNA repair in Archaea
Authors: Dantuluri Swathi , Margulies Shae , Hepowit Nathaniel , Allers Thorsten , Maupin‐Furlow Julie 
Published: June 2021

Cdc48/p97/VCP is an AAA ATPase, conserved in eukaryotes and archaea, which is demonstrated to convert chemical energy from the hydrolysis of ATP into the mechanical energy, for either unfolding protein substrates or dislodging protein substrates from complexes. The role of Cdc48 in ubiquitin mediated protein degradation has been well reported; however, its role in DNA repair is less understood so far. Evidence in eukaryotes which implicates Cdc48 in DNA repair is based on the association of Cdc48 with partners and substrate proteins, which are known to be involved in DNA damage response. In our study, we found Cdc48A of Haloferax volcanii co‐purifies with RecJ‐domain (RecJ3 and RecJ4) and RNase J1 homologs in an apparent thiol sensitive complex that binds the oxidative stress‐responsive, ubiquitin‐like SAMP1 in the presence of ATP. RecJ domain proteins are DNases reported to be involved in DNA repair pathways in bacteria. Further analysis revealed an Hfx. volcanii Δcdc48A mutant showed significant difference in survival when compared to its wildtype, on subjecting them to DNA damaging agents like phleomycin and UV. An increase (up to 3.4‐fold; p < 0.05) in transcript levels of cdc48a, recj3 and rnj1 was observed after treatment with the DNA damaging agent phleomycin, suggesting the expression of these genes is correlated with DNA repair. Current focus of this study is to decipher the role of Cdc48A in DNA repair by identifying its partner and substrate proteins. In addition, we are determining the substrate modifications required for signaling the Cdc48A mediated repair pathway using classic biochemistry techniques and genetic manipulations.Support or Funding InformationThe work was supported by funds awarded to Dr. Julie A. Maupin‐Furlow through the Bilateral NSF/BIO‐BBSRC program (NSF 1642283), U.S. Department of Energy, Office of Basic Energy Sciences, Division of Chemical Sciences, Geosciences and Biosciences, Physical Biosciences Program (DOE DE‐FG02‐05ER15650) and the National Institutes of Health (NIH R01 GM57498).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protein myristoylation links a high fat diet with prostate tumor progression in mice
Authors: Cai Houjian , Kim Sunjin 
Published: June 2021

Objective of the studyExogenous fatty acids provide substrates for energy production and the biogenesis of the cytoplasmic membrane, but they also enhance cellular signaling during cancer cell proliferation. However, it remains controversial whether dietary fatty acids are correlated with tumor progression. Additionally, the molecular mechanisms explaining the effect of dietary fatty acids in promoting prostate cancer progression remains unknown.Statement of MethodsIn this study, prostate regeneration assay and prostate cancer cell xenografts were applied to study if and how a high fat diet promotes tumor progression in vivo. The amount of Acyl‐CoAs were analyzed by LC–MS/MS analysis. Myristoylation of Src kinase was detected by Click chemistry. Additionally, to examine the molecular mechanisms, proteins were fractionated into cytosol and membrane fraction. Expression levels of Src kinase were detected by immunoblotting.Summary of the resultsWe demonstrate that increased Src kinase activity is associated with high fat diet accelerated progression of prostate tumors and that Src kinases mediate this pathological process. Moreover, in the in vivo prostate regeneration assay, host SCID mice carrying Src(Y529F) transduced regeneration tissues were fed with a low fat diet or a high fat diet and treated with vehicle or dasatinib. The high fat diet not only accelerated Src induced prostate tumorigenesis in mice, but also compromised the inhibitory effect of the anticancer drug dasatinib on Src kinase oncogenic potential in vivo. We further show that myristoylation of Src kinase is essential to facilitate Src induced and high fat diet accelerated tumor progression. Mechanistically, metabolism of exogenous myristic acid increased the biosynthesis of myristoyl CoA and myristoylated Src, and promoted Src kinase mediated oncogenic signaling in human cells. Of the fatty acids tested, only exogenous myristic acid contributed to increased intracellular myristoyl‐CoA levels.ConclusionOur results indicate that targeting Src kinase myristoylation, which is required for Src kinase association at the cellular membrane, blocks dietary fat accelerated tumorigenesis in vivo. Our findings uncover a molecular basis, which the biosynthesis of myristoyl‐CoA elevates the expression of myristoyl‐Src, and accelerates Src mediated prostate tumor progression in a high fat diet.Support or Funding InformationNIH (R01CA172495) and DOD (W81XWH‐15‐1‐0507)
Targeting N‐myristoyltransferase1 inhibits prostate cancer progression
Authors: Alsaidan Omar Awad , Kim Sungjin , Li Qianjin , Bielawska Alicja , Cai Houjian 
Published: June 2021

N‐myristoyltransferase (NMT) facilitates co/post‐translational myristoylation of several oncogenic proteins, regulating their functions in tumor progression. Overexpression of N‐myristoyltransferase (NMT), which has been shown to be upregulated in a variety of cancers, is associated with androgen independent prostate cancer cells. We demonstrate that genetically ablation of NMT1 inhibited proliferation of prostate cancer cells, tumor growth, and suppressed myristoylation profile of prostate cancer cells. Screening a panel of myristoyl‐CoA analogs against purified human NMT1 protein lead to identifying the B13/LCL4 as an inhibitor for NMT1. B13/LCL4 significantly supressed prostate cancer cell proliferation, migration, and invasion by cell cycle arrest, and inhibited the growth of prostate xenograft tumors with minimal pathological effect on major organs in vivo. Structure activity relationship based optimization of B13 led to LCL204, which showed better inhibitory properties twoards NMT1. We demonstrate that targeting protein myristoylation is a potential therapeutic approach to inhibit prostate tumor progression.Support or Funding InformationThis work was supported by NIH (R01CA172495) and DOD (W81XWH‐15‐1‐0507) to H. CaiThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Detrimental Effects of Aging, Ovariectomy and Weight Gain on Left Ventricular Structure and Function: A Potential Preclinical Model of Early Stage HFpEF
Authors: Bustamante Moises , Garate‐Carrillo Alejandra , Loredo Maria , Garcia Ricardo , Carson Nancy , Ito Bruce , Ceballos Guillermo , Omens Jeffrey , Ramirez‐Sanchez Israel , Villarreal Francisco 
Published: June 2021

The incidence of heart failure with preserved ejection fraction (HFpEF) continues to increase and little is known about its pathophysiology. Two‐thirds of HFpEF patients are senile women and risk factors include hypertension and metabolic syndrome. Currently, no drugs are effective for HFpEF treatment and the development of animal models would assist in novel therapy evaluation. We implemented a rodent female animal model of aging, estrogen depletion and weight gain to evaluate the role of these factors in altering cardiac structure/function. Female, 18 month old, Fischer F344 rats were allocated into an aging (A) group, aging + ovariectomy (O) and aging + ovariectomy and 10% fructose (OF) in drinking water (n=8–13/group). For comparison purposes a young (3 month old, n=8) cohort of female rats was included. Left ventricular (LV) structure/function was monitored monthly by echocardiography. At 22 months of age, animals were anesthetized and LV function evaluated using a Millar conductance catheter. Histological measures of chamber morphometry, inflammation and collagen density (fibrosis) were also obtained. OF animals significantly increased body weight vs. A and O. While echocardiography did not detect major alterations in cardiac endpoints, intraventricular pressure‐volume loop analysis evidenced significant (p<0.05) decreases in stroke volume (19% O and 27% for OF vs. A), stroke work (27 O and 31% OF vs. A), cardiac output (43 O and 44% OF vs. A) and increases in Tau (30–35% in A, O and OF vs. young) and LV end‐diastolic pressure volume relationship slope (i.e. stiffness) with ovariectomy in the presence of preserved ejection fraction for all groups. Histomorphometry did not detect changes in chamber dimensions. However, histological analysis indicated progressively increasing levels of inflammatory infiltration, perivascular and interstitial fibrosis with ovariectomy and with fructose supplementation in particular, in the subendocardium (see figure). In conclusion, in the setting of aging, ovariectomy (which depletes estrogens), further deteriorates myocardial microstructure which may facilitate the loss of diastolic and global functional indices in the apparent presence of preserved ejection fraction. This model may serve to understand the role that estrogen depletion and weight gain may have in the development of HFpEF.Support or Funding InformationDoD 150090 to FVThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
History‐Dependence of the Force‐Velocity Relationship in mdm Mouse Muscles
Authors: Piwinski Lindsay , Rice Nicole , Tahir Uzma , Monroy Jenna , Nishikawa Kiisa 
Published: June 2021

The force generated by a muscle is inversely related to the velocity of the shortening. Maximum shortening velocity (Vmax) has been predicted from highly controlled, isotonic conditions which are rarely encountered in vivo. During natural movements, muscles undergo stretch‐shortening cycles, which alter power output and predict higher Vmax than traditional isotonic experiments. We investigated the mechanism underlying the differences in the FV relationship. We hypothesized that for a given force, Vmax is greater during work loop experiments because during stretch a muscle can store elastic energy that can be recovered upon shortening. It is possible that the giant elastic protein, titin, functions as a spring in active muscle and therefore contributes to the storage and recovery of elastic energy during work loops but not isotonic contractions. To test these hypotheses, we compared the FV relationships of soleus muscles from wild type and mdm mice with a deletion in the N2A region of titin. Results showed that Vmax is lower under isotonic conditions compared to work loops in both genotypes but the difference was more than 20% greater in wild type muscles. Thus, titin, cross‐bridge kinetics or some combination of both may explain the observed differences. This work contributes to a growing body of evidence of titin's potential role as a spring in active muscle and sheds light on the underlying mechanism of active muscle stiffness.Support or Funding InformationNSF IOS‐1456150 awarded to J.Monroy, APS UGSRFThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Liposomal dexamethasone attenuates tourniquet‐induced ischemia‐reperfusion injury in mouse hindlimb
Authors: Tu Huiyin , Zhang Dongze , Wadman Michael C. , Li Yu‐Long 
Published: June 2021

Tourniquet‐induced ischemia and subsequent release cause serious ischemia‐reperfusion (IR) injury in the skeletal muscle. Here, we observed the effect of liposomal dexamethasone (lipo‐Dex) in tourniquet‐Induced ischemic reperfusion injury (IR) in mouse hindlimb.Unilateral hindlimb of C57/BL6 mice was subjected to 3 hours of ischemia by placing a rubber band and followed different periods of reperfusion (1 week, 2 weeks, 4 weeks, and 6 weeks). Before tourniquet was released, lipo‐Dex (7 mg/kg, equivalent to amg/kg/day of Dex for 1 week) was administered via tail vein injection. After 3 hours of ischemia and different periods of reperfusion, gastrocnemius muscle showed significantly morphological and functional damages. TNFα and IL‐1β were overexpressed in the gastrocnemius muscle after 1, 2, 4, and 6 weeks of tourniquet‐induced IR. Treatment with Lipo‐Dex inhibited the elevation of TNFα and IL‐1β, improved structure of the muscle sarcolemma, reduced fragmentation of nicotinic acetylcholine receptor clusters, and promoted recovery of gastrocnemius muscle contraction.The data suggest that tourniquet‐related IR induces serious skeletal muscle inflammation and degradation. Treatment with lipo‐Dex suppresses IR‐induced inflammation and promotes recovery of skeletal muscle morphology and function from tourniquet‐induced IR injury.Support or Funding InformationUSAMRAA grant from Department of Defense, W81XWH‐17‐1‐0037(PI, Yulong Li).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.