bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cocaine Enhances the Discriminative Stimulus Effects of Mephedrone
Authors: Erwin Laura , Jursic Branko , Winsauer Peter 
Published: June 2021

Mephedrone (4‐methylmethcathinone) is one of the major constituents of “bath salts.” It is a drug with discriminative effects similar to CNS stimulants and it can produce auditory and visual hallucinations, as well as problematic cardiovascular effects. This study compared the discriminative stimulus effects of mephedrone (0.32–10 mg/kg) with other prototypical drugs with CNS effects, such as ketamine (1.8–18 mg/kg), bupropion (5.6–56 mg/kg), and 3,4‐methylenedioxyamphetamine (MDA) (0.32–5.6 mg/kg). In addition, mephedrone was administered in combination with cocaine (5.6–32 mg/kg), which inhibits the reuptake of catecholamines and serotonin. Rats (n=6) were trained to discriminate an intraperitoneal injection of 3.2 mg/kg mephedrone from saline under a fixed‐ratio 20 schedule. Following training, increasing cumulative doses of mephedrone produced dose‐dependent increases in mephedrone‐lever responding, with full substitution considered to be greater than 80% responding and partial substitution to be greater than 50% responding. During substitution tests, ketamine never produced more than 17% drug‐lever responding up to doses that significantly decreased overall response rate. In addition, bupropion and MDA produced 36% and 77% drug‐lever responding, respectively, without significantly decreasing response rate. When 3.2 mg/kg of cocaine was administered prior to increasing doses of mephedrone, there was no substitution for cocaine alone; however, this dose produced a rightward shift in the dose‐effect curve for mephedrone‐lever responding. Unlike this dose, 5.6 mg/kg of cocaine alone did not substitute for mephedrone or shift the mephedrone dose‐effect curve. When the dose of cocaine was increased to 10 mg/kg, there was still no mephedrone substitution, but this dose increased the mephedrone‐lever responding of the low doses of mephedrone. Finally, 18 and 32 mg/kg of cocaine produced 38.1% and 77.7% substitution, respectively, and dose‐dependently increased the substitution of the low doses of mephedrone (0.32 – 1.8 mg/kg). This data suggest that mephedrone may be similar to cocaine in having a higher affinity for the serotonin transorter (SERT) than the dopamine transporter (DAT), but more efficacy at DAT than SERT. Certainly, due to the capacity of cocaine to only enhance the substitution of the low mephedrone doses, the discriminative stimulus effects of the training dose of mephedrone are mediated predominately by dopamine rather than serotonin.Support or Funding InformationThis research was supported by the Louisiana Board of Regents Fellowship.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 effect of Porphyromonas gingivalis Lipopolysaccharide (LPS) on Rat Brain Microglia Classical and Alternative Activation in vitro: MMP‐9 generation
Authors: Czerwonka Evan , Memedovski Zylfi , Han Jin , Mayer Joshua , Klemm Lucas C. , Hall Mary L. , Mayer Alejandro M. 
Published: June 2021

BackgroundThe putative association between periodontal disease (PD), a chronic inflammatory disease affecting 64% of adults 65 years and older, and neuroinflammation, a condition which involves microglia, a cell of the brain innate immune system, remains relatively unexplored. We recently reported that lipopolysaccharide (LPS) of Porphyromonas gingivalis (Pg), a pathogenic bacteria implicated in periodontitis activates both classical (M1‐type) and alternative (M2‐type) activation of neonatal rat microglia (BMG) and concomitant release of superoxide anion, thromboxane B2, cytokines, and chemokines (Toxicological Sciences Supplement, 150 (1), Abstract # 2974, 2017). We hypothesized that classical activated microglia might also release the pro‐inflammatory matrix metalloproteinase‐9 (MMP‐9).MethodsP. gingivalis LPS (Pg LPS) was purchased from InvivoGen (San Diego, CA). E. coli LPS (Ec LPS) 026:B6 (from Difco Lab, Detroit, MI) was used as a positive control. BMG were isolated from neonatal rats, and treated in vitro with either Pg LPS or Ec LPS, in a concentration‐dependent manner, for 18 hours at 35.9°C. BMG were identified by confocal microscopy using the surface marker CD11b/c. Matrix metalloproteinase‐9 was determined with a rat total MMP‐9 DuoSet ELISA kit (R&D Systems).ResultsStatistically significant and concentration‐dependent release of pro‐inflammatory MMP‐9 was observed at > than 1ng/mL Ec LPS and 105 ng/mL Pg LPS.ConclusionsOur results provide support for our working hypothesis, because Pg LPS stimulated BMGs to release pro‐inflammatory MMP‐9 in a time‐ and concentration‐dependent manner. Thus, our current observations extend our studies on the effect of Pg LPS on rat brain microglia activation and contribute to further characterize both periodontitis, and the role of BMG in neuroinflammation and potential neurodegeneration.Support or Funding InformationSupport by College of Dental Medicine, Midwestern University is gratefully acknowledged.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Manganese Exposure Activates NLRP3 Inflammasome Signaling and Propagates Exosomal Release of ASC in Microglial Cells
Authors: Malovic Emir , Sarkar Souvarish , Rokad Dharmin , Luo Jie , Harischandra Dilshan , Jin Huajun , Anantharam Vellareddy , Huang Xuemei , Lewis Mechelle , Kanthasamy Arthi , Kanthasamy Anumantha 
Published: June 2021

Abnormal metal homeostasis is a key hallmark of multiple neurodegenerative disorders including Alzheimer's and Parkinson's diseases. Toxicological exposure to manganese (Mn) has been linked to PD‐like symptoms. Sustained neuroinflammation is now recognized as a key pathophysiological process of neurodegenerative conditions, but the inflammatory signaling mechanism underlying Mn neurotoxicity remains equivocal. Recent evidence suggests that the NLRP3 inflammasome, a multiprotein complex, can be activated by various exogenous and endogenous stimuli, leading to a chronic inflammatory response. In this study, we examined whether Mn exerts its neurotoxic effect by activating NLRP3 inflammasome signaling. Exposing LPS‐primed microglial cells to Mn significantly increased NLRP3 expression, caspase‐1 cleavage, and IL‐1β maturation, indicating that Mn can induce the NLRP3 inflammasome activation cascade. Mn exposure compromised the mitochondrial dynamics of microglial cells, as measured via Seahorse Mito Stress assay, and downregulated mitochondrial fusion. Furthermore, Mn reduced the expression of the retromer pathway protein VPS35. CRISPR/Cas9‐based knockdown of VPS35 also downregulated the mitochondrial fusion protein Mfn2 by increasing the mitochondrial ubiquitin ligase Mul1, suggesting a probable role of VPS35 in maintaining mitochondrial health. Given that the inflammasome component ASC can propagate inflammasome activation in a prionic manner, we explored whether Mn exposure induces cell‐to‐cell transfer of ASC via exosomes. Exosomes isolated from Mn‐treated, LPS‐primed microglial cells contained higher amounts of ASC, suggesting exosomal release of ASC. Supporting the role of exosomal ASC in inflammasome propagation, we found that exosomes isolated from microglial cells exposed to Mn were able to induce pro‐IL‐1β and NLRP3 expression. Importantly, we confirmed a significantly higher ASC content in exosomes collected from Mn‐exposed welders when compared to exosomes from matched control subjects. Collectively, these results demonstrate that Mn activates NLRP3 inflammasome proinflammatory signaling and exosomal ASC release from microglia to induce neurotoxicity.Support or Funding InformationES026892, NS088206, ES027245, and Endowed Lloyd ChairThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EFFECT OF VESSILEN®, A NEW FORMULATION OF ADELMIDROL AND SODIUM HYALURONATE, ON OVERACTIVE URINARY BLADDER SYNDROME
Authors: Cuzzocrea Salvatore , Peritore Alessio , Crupi Rosalia , Campolo Michela , Siracusa Rosalba , Cordaro Marika , Filippone Alessia , Esposito Emanuela 
Published: June 2021

The disruption of coordinated control between the brain, spinal cord and peripheral nervous system caused by spinal cord injury leads to several secondary pathological conditions, including lower urinary tract dysfunctions. In fact, a disability after SCI is urinary dysfunction, which could be attributable to lack of neuroregeneration of supraspinal pathways that control the bladder function. As a consequence, individuals develop overactive urinary bladder (OAB). The aim of the current study was to explore the effects of vessilen®, a new formulation of 2% adelmidrol + 0.1% sodium hyaluronate, in OAB generated after SCI in mice. Mice were treated with vessilen® (100ml/mouse, intravesical) daily for 48 h and 7 days after SCI. Vessilen® reduced significantly mast cells degranulation and down regulated NFκB pathway in the bladder after SCI both at 48h and 7 days. Moreover vessilen® was able to reduce NGF expression suggesting an association between neurotrophins and bladder pressure. Moreover, 7 days after SCI, bladder was characterized by a considerable bacterial infection and proteinuria, surprisingly vessilen® reduced significantly both parameters. These data show the protective roles of vessilen® in OAB following SCI, highlighting a potential therapeutic target for controlling OAB, making a significant impact on improving overall quality of life.Support or Funding Informationno fundingThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hyperlocomotion induced by inhalation of nicotine and a low dose of Δ9‐Tetrahydrocannabinol in female rats
Authors: Paydar Mehrak Javadi , Vandewater Sophia A. , Taffe Michael Javadi 
Published: June 2021

BackgroundsGrowing availability of e‐cigarette devices permits users to prepare their own cocktail of drugs for inhalation, including polysubstance inhalation. Some individuals use nicotine and (−)‐Δ9‐tetrahydrocannabinol (THC) in combination by smoking tobacco and cannabis together or in sequence; this practice may extend to electronic cigarette users. THC exerts biphasic effects on locomotion in rats, with high doses (e.g., 5 mg kg−1, i.p.) decreasing, and lower doses (e.g., 0.03 mg kg−1, i.p.) increasing, locomotor activity. Nicotine (NIC) also increases locomotor activity in rodents. This study was designed to determine if interactive effects of THC and NIC result from vapor inhalation in rats using a novel e‐cigarette based procedure.MethodsFemale Sprague‐Dawley rats were exposed to vapor produced by the propylene glycol vehicle (PG), NIC (1 mg/mL in PG), THC (12.5 mg/ml in PG) or a combination of NIC with THC for a duration of 15 minutes. Body temperature and locomotor responses were evaluated post‐inhalation using a radiotelemetry system.ResultsLocomotor activity was increased in the rats for the first 60 min after NIC (1 mg/ml) exposure and this effect was blocked by pretreatment with mecamylamine. Activity was also increased 15 min after initiation of THC vapor compared with PG. Female rats did not show any further hyperlocomotion following exposure to the combination of NIC+THC vapor compared with either NIC or THC vapor inhalation, alone. The body temperature was decreased following the THC (12.5 mg/ml) or a combination of THC (12.5 mg/ml) +NIC (1 mg/ml) inhalation with significant effects starting 120 min post‐exposure, while NIC (1 mg/ml) inhalation did not alter the body temperature.DiscussionVapor inhalation of NIC or low dose THC stimulates locomotor activity in female rats. The co‐administration of a lower dose of THC with NIC produced no additive locomotor stimulation in female rats compared to either THC or NIC alone. The observed data provided behavioral evidence to suggest that the effect of nicotine on locomotion might not be modified by the physiological action of cannabinoids.Support or Funding InformationFunding support provided by USPHS grants AA007456, DA035482 and DA041967This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification and Characterization of ML321: A Novel and Selective D2 Dopamine Receptor Antagonist with Predicted Atypical Antipsychotic Properties
Authors: Free R. Benjamin , Xiao Jingbo , Ferrer Marc , Southall Noel , Mach Robert , Bertz Jeremiah , Shi Lei , Woods James H. , Marugan Juan J. , Wetsel William , Sibley David R. 
Published: June 2021

Dopamine receptors (DARs) belong to the G protein‐coupled receptor (GPCR) superfamily, whose members play critical roles in cellular signaling, especially modulating the transfer of information within the nervous system. Amongst DARs, the D2 dopamine receptor (D2R) is one of the most validated drug targets in neuropsychiatry and there is a strong correlation between the clinical doses of antipsychotics and their potencies for blocking D2Rs. However, current FDA‐approved antipsychotic drugs typically cross‐react with other GPCRs, leading to many serious off‐target side‐effects. A potent and highly selective D2R antagonist would thus be extremely valuable for the treatment of psychosis and related CNS disorders. Using high throughput screening techniques, we have identified, optimized, and characterized a lead D2R antagonist with high selectivity – ML321. In a functional profiling screen of 168 different GPCRs, ML321 showed relatively little reactivity beyond inhibition of the D2R, and to a lesser extent the D3R, demonstrating exceptional GPCR selectivity. Interestingly, we found that the binding of ML321 to the D2R is entirely dependent on Na+. PET imaging studies in non‐human primates demonstrated that ML321 can penetrate the CNS and occupy the D2R in a dose‐dependent manner. Preliminary in vitro ADME studies showed that ML321 is metabolically stable and did not block hERG channels, whereas pharmacokinetic studies in mice demonstrated elevated plasma and brain levels of ML321 for up to 8 hrs. Behavioral paradigms in rats demonstrated that that ML321 can selectivity antagonize a D2R‐mediated response (hypothermia) while not affecting a D3R‐mediated response (yawning) using the same dose of drug, thus demonstrating good in vivo selectivity. We also investigated the effects of ML321 in animal models that are predictive of antipsychotic efficacy in humans. We found that ML321 can attenuate both PCP and amphetamine‐induced locomotor activity and pre‐pulse inhibition (PPI) in a dose‐dependent manner. Importantly, using doses that were maximally effective in the locomotor and PPI studies, ML321 promoted little‐no catalepsy compared with the nonselective antipsychotic haloperidol. This latter observation suggests that ML321 may produce fewer extrapyramidal (motor) side‐effects, a common problem with FDA‐approved antipsychotics. Overall, these results suggest that the ML321 scaffold could serve as a lead compound for the development of an improved therapeutic with greatly reduced side‐effects for treating schizophrenia and other psychotic syndromes.Support or Funding InformationNINDS IRPThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fibroblast Growth Factor Binding Protein 3 A Novel Target for Glucose Intolerance and Nonalcoholic Fatty Liver Disease Treatment
Authors: Garman Khalid , Tassi Elena , Schmidt Marcel , Wellstein Anton 
Published: June 2021

Endocrine Fibroblast Growth Factor 21 (FGF21) has been shown to improve insulin sensitivity, glucose tolerance, and hepatosteatosis in many species. Previous studies have demonstrated that Fibroblast Growth Factor Biding Proteins (FGFBPs) protects FGFs from proteolytic degradation and enhance their binding to their cognate receptors. In our studies, we report that FGFBP3−/− mice suffer insulin resistance and impaired glucose homeostasis, which was further aggravated by diet‐induced obesity. In FGFBP3−/− mice, we further report on the scant liver fat content in lean regular diet mice and the paradoxical high‐fat diet‐induced hepatosteatosis. We show that endogenous FGFBP3 controls liver fat content through its effect on fatty acid synthesis (Fasn, ACAC, ACLY, AACS, ACAS2, and SCD1), fatty acid uptake (SLC27A5), long‐chain fatty acid activation (ACSL1 and ACSL5), and triglyceride synthesis (Dgat1 and Dgat2). In contrast, we demonstrate that exogenous FGFBP3 administration ameliorates hepatosteatosis in ob/ob mice via promoting peroxisomal fatty acid oxidation (EHHADH) and triglyceride secretion (MTTP). We also describe the high‐fat diet‐induced upregulation of hepatic and white adipose tissue FGF21 and KL® in the FGFBP3−/− mice. Our studies indicate a more prominent FGFBP3 effect on the liver compared to white adipose tissue. Collectively, our studies indicate a possible collaboration between FGFBP3 and FGF21 on blood glucose and liver fat content homeostasis.Support or Funding InformationSupported by NIH grants P01 HL068686 (CSW, AW), R01 CA71508 (AW), P30 CA51008 (AW)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exploring the Mechanisms by which Small Molecule Nef Inhibitors Suppress HIV‐1 Infectivity
Authors: Staudt Ryan P. , Emert‐Sedlak Lori , Smithgall Thomas E. 
Published: June 2021

Nef is an HIV‐1 accessory factor essential for viral pathogenesis and immune escape of HIV‐infected cells. Recently, our group reported the discovery of small molecule Nef antagonists that reverse Nef‐mediated downregulation of MHC‐I and restore the anti‐HIV cytotoxic T cell response. These compounds also inhibit Nef's ability to enhance the inherent infectivity of HIV‐1 virions, although their mechanism of action in this regard is not known. Recent studies have identified the host cell factor SERINC5 (S5) as a suppressor of HIV‐1 infectivity that is countermanded by Nef. Here we investigated whether our Nef inhibitors suppress viral infectivity through a S5‐dependent mechanism. We first confirmed that expression of S5 within viral producer cells results in Nef‐reversible inhibition of HIV‐1 infectivity. For these experiments, we co‐transfected 293T cells with wild‐type and ΔNef HIV‐1 proviral DNA along with various amounts of a S5 expression plasmid. The infectivity of each resulting virus was then quantified using the luciferase‐based HIV‐1 reporter cell line, TZM‐bl. As expected, co‐expression of S5 resulted in suppression of HIV‐1 infectivity. This S5 effect was reduced by Nef in cells producing wild‐type HIV‐1, but not in cells producing the ΔNef virus. Co‐expression of Nef from a separate expression plasmid during the production of the ΔNef virus completely reversed S5 inhibition of viral infectivity, demonstrating Nef dependence in this system. To determine whether our inhibitors block Nef‐enhanced infectivity in a S5‐specific manner, we treated cells producing wild‐type or ΔNef virus in the presence or absence of S5 with either DMSO or the diphenylpyrazolo Nef inhibitor, B9 (3 μM). This compound inhibited the infectivity of wild‐type but not ΔNef HIV‐1 at the producer cell level, consistent with direct action on Nef. However, the effect of the compound was independent of S5 expression, suggesting that mechanisms in addition to S5 may contribute to inhibition of HIV‐1 infectivity by B9 through Nef.Support or Funding InformationT32 AI65380 (R.P.S.)R01 AI057083, R42 GM112516 (T.E.S.)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mitophagy is Required for Proper Mitochondrial Network Remodeling during Cardiac Progenitor Cell Differentiation
Authors: Lampert Mark , Orogo Amabel , Najor Rita , Gustafsson Åsa 
Published: June 2021

Cell based therapies represent a very promising strategy to repair and regenerate the injured heart to prevent progression to heart failure. To date, cell based therapies have had limited success due to a lack of survival and retention of the infused cells. Therefore, it is important to increase our understanding of the biology of these cells and utilize this information to enhance their survival and function in the unfavorable environment of the injured heart. Mitochondria are critical for progenitor cell function and survival. Here we demonstrate the importance of mitochondrial autophagy, or mitophagy, in the differentiation process in adult cardiac progenitor cells (CPCs). We found that mitophagy is rapidly induced upon initiation of differentiation in CPCs. We also found that mitophagy was mediated by the mitophagy receptors pathway, rather than the PINK1/Parkin pathway. Mitophagy receptors Nix and Fundc1, but not Bnip3, were upregulated during differentiation. Mitophagy mediated by Nix and Fundc1 was not involved in regulating progenitor cell fate determination, mitochondrial biogenesis, or reprogramming. Instead, mitophagy facilitated the CPCs to undergo proper mitochondrial network reorganization during differentiation. Abrogating Nix/Fundc1‐mediated mitophagy during differentiation led to mitochondrial fragmentation and failure to form an interconnected mitochondrial network. It also led to increased susceptibility to hydrogen peroxide mediated cell death during differentiation. Finally, aging is associated with accumulation of mtDNA mutations in cells and we found that acquiring mtDNA mutations selectively disrupted the differentiation‐activated mitophagy program in CPCs. These findings demonstrate the importance of Nix/Fundc1‐mediated mitophagy as a critical regulator of mitochondrial network formation in differentiating progenitor cells, as well as the consequences of accumulating mtDNA mutations.Support or Funding InformationA.B. Gustafsson is supported by an AHA Established Investigator Award, and by NIH R21AG052280, R01HL087023, R01HL132300 and P01HL085577. M.A. Lampert is supported by the UCSD Graduate Training Program in Cellular and Molecular Pharmacology grant T32GM007752. A.M. Orogo is supported in part by the UCSD Graduate Training Program in Cellular and Molecular Pharmacology through an institutional training grant from the National Institute of General Medical Sciences T32GM007752, and National Institutes of Health NRSA Predoctoral Fellowship F31HL123309.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.