bluuebunny/crossref_metadata_2025_split huggingface dataset:
Claudin‐5 as a target for drug delivery to the brain by modulation blood‐brain barrier
Authors: Kondoh Masuo , Hashimoto Yosuke , Shirakura Keisuke , Okada Yoshiaki , Takeda Hiroyuki , Endo Kohki , Tamura Maki , Watari Akihiro , Sadamura Yoshifusa , Sawasaki Tatsuya , Tachibna Keisuke , Doi Takefumi , Yagi Kiyohito 
Published: June 2021

The regulatory success rate for central nervous system (CNS) drugs is less than half that of non‐CNS drugs. A current bottleneck in the development of CNS drugs is the lack of drug delivery systems to the brain. The blood‐brain barrier (BBB) is a membrane barrier formed by brain capillary endothelial cells and protects the brain from harmful materials circulating in the blood. Most small and all large drugs do not readily cross the BBB. The intercellular space between endothelial cells of the BBB is sealed by complex protein‐based structures called tight junctions (TJs). Claudin‐5 (CLDN‐5), a tetra‐transmembrane protein, is a key component of the TJ seal that prevents the paracellular diffusion of drugs into the CNS. Here, to investigate whether CLDN‐5 can be a target for CNS delivery of drugs, we generated monoclonal antibodies (mAbs) specific to the extracellular domain of CLDN‐5 using DNA immunization. Treatment of MDCKII/CLDN‐5 cells with anti‐CLDN‐5 mAbs time‐ and dose‐dependently decreased the integrity of TJs. The integrity was restored by removal of the anti‐CLDN‐5 mAbs. In an in vitro model of the BBB, anti‐CLDN‐5 mAbs also attenuated the integrity of TJs and enhanced permeability of small (Mw 376 Da) and large (4 kDa) solutes. The anti‐CLDN‐5 mAbs did not decrease TJ integrity in human intestinal epithelium models. In conclusion, CLDN‐5 is a potential target for the development of drug delivery systems to the brain. The anti‐CLDN‐5 mAb is a potential lead compound for development of novel drug delivery systems targeting the CNS.Support or Funding InformationThis study was partly supported by Grant‐in‐Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (18K19400, 18H03190, 16K13044).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Honokiol Attenuates Amyloid‐beta Induced Neuroinflammatory Microglial Polarization in BV2 Microglial Cells
Authors: Jones Ellery , Govindarajulu Manoj , Ramesh Sindhu , Dhanasekaran Murali 
Published: June 2021

Neuroinflammation plays a crucial role in the pathogenesis of Alzheimer's Disease (AD). Microglia are the macrophages of the central nervous system (CNS), which play a crucial role in immune surveillance and maintaining homeostasis. Chronic microglial activation is detrimental because they produce several pro‐inflammatory cytokines, which result in neuronal cell death (M1 phenotype). Additionally, several studies indicate that anti‐inflammatory mechanisms are down‐regulated in AD (M2 phenotype). Honokiol is a lipophilic polyphenol known to possess potent anti‐inflammatory properties. The goal of this study was to determine the effect of honokiol on activated microglia. The microglial cell line BV2 was pretreated with Honokiol, followed by treatment with amyloid beta, for 24 hours. Pro‐inflammatory activation was assessed by measuring nitric oxide, TNF‐α and IL‐6 levels. Biochemical assays, including western blot, immunocytochemistry and PCR, were performed to mechanistically determine the effect of honokiol on microglial activation. Honokiol significantly reduced NO, ROS, TNF‐α and IL‐6 levels. Honokiol also significantly reduced iNOS expresssion, down‐regulated markers of M1 phenotype and increased M2 markers. The promotional effects of Honokiol on M2 polarization were attenuated by an AMP‐activated protein kinase (AMPK) inhibitor, indicating that AMPK activation participates in Honokiol's effects. Furthermore, Honokiol‐mediated M2 gene expression was prevented by an inhibitor of calmodulin‐dependent protein kinase kinase β (CaMKKβ), an upstream kinase of AMPK. Our in vitro studies indicate that Honokiol could be acting as an AMPK activator, thereby enhancing M2 polarization and suppressing microglial activation. Further studies in animal models need to be conducted to validate Honokiol as a novel and safe therapeutic compound for the treatment of neuroinflammation associated with AD.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Monitoring Signalling and Trafficking of Neurotensin Type 1 Receptor in Animal Model using Fluorescent‐based Methods
Authors: Couvineau Pierre , Ehrlich Aliza , Semache Meriem , Kieffer Brigitte , Bouvier Michel 
Published: June 2021

Neurotensin Type 1 Receptor (NTSR1) is a G Protein‐Coupled Receptor (GPCR) with proposed roles in cancer, neuropsychiatric and neurodegenerative diseases, eating disorders, pain, as well as inflammation. It therefore represents a potential therapeutic target for the treatment of these diseases. However, relatively little is known about the repertoire of signalling effectors that can be engaged by NTSR1 and about the functional selectivity of drug candidates targeting this receptor in native tissue.In a first step toward generating an animal model allowing to monitor the activation and regulation of NTSR1 in native tissues, we developed a DNA construct expressing NTSR1 tagged to vYFP at its C‐terminus.We investigated the signalling responses of NTSR1‐vYFP promoted by eight agonists and two antagonists, and compared them to those obtained for the unmodified NTSR1. For this purpose, we used a suite of BRET‐based sensors expressed in HEK‐293 cells. We showed that NTSR1‐vYFP has the same signalling profile as the endogenous NTSR1 as it activates both β‐Arrestin 1 and 2 and all G proteins except G12 and Gi3. Moreover, we confirmed that ML314 is a biased agonist toward β‐Arrestin pathways and we showed that PD149163 does not activate Gs protein.Because the vYFP fusion does not seem to modify the signalling profile of the NTSR1, we have embarked on developing a knock‐in mouse model expressing NTSR1‐vYFP instead of the native receptor which will be a useful tool to study receptor signaling and trafficking under physiological conditions.Support or Funding InformationThe project is funded by the Consortium de recherche biopharmaceutique (CQDM). Pierre Couvineau is funded by the INSERM‐FRQS Postdoctoral Exchange ProgramThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of different adenylyl cyclase isoforms impacts ligand bias downstream of the kappa opioid receptor
Authors: Brust Tarsis , Scarry Sarah , Aubé Jeffrey , Bohn Laura 
Published: June 2021

Functionally selective (or biased) ligands have been widely pursued in recent years as a way to target G protein‐coupled receptors (GPCRs) in a more selective manner. The promise of biased signaling is that by specifically activating only a subset of a receptor's responses, therapeutic effects can be achieved and adverse effects prevented. Molecular studies of biased ligands are commonly carried out in cell lines, and many of them use cyclic AMP accumulation as a readout of G protein activation. The production of cyclic AMP is catalyzed by adenylyl cyclases (ACs). In humans there are 10 different isoforms of ACs, which diverge in their expression patterns and regulatory properties. In the present study, we examined if the expression of different AC isoforms in a cell line could result in different bias profiles for ligands of the kappa opioid receptor (KOR). We employed one biased and two reference ligands and showed that there were significant changes in potency as well as in bias factors depending on the AC isoform that was expressed in our cellular system. Our results support the idea that selection of an appropriate cellular model is an invaluable step in studies of functional selectivity. Moreover, using cells that match the AC expression pattern of the tissue where the compound is predicted to act can lead to more relevant results when cyclic AMP is used as a readout of G protein activation.Support or Funding InformationThis work was supported by National Institute on Drug Abuse grant R01DA031927.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Aldosterone binding to G Protein Estrogen Receptor‐GPER
Authors: Ding Qingming , Chorazyczewski Jozef , Gros Robert , Feldman Ross 
Published: June 2021

MR‐independent effects of aldosterone acting through GPER have been appreciated since 2011. However, whether this related to direct aldosterone‐GPER binding has been disputed. Two previous radioligand studies failed to demonstrate direct aldosterone binding to a putative GPER. However, in neither study was it demonstrated that the radioligand used, bound to a physiological receptor. Further, both utilized 3H estradiol‐ whose low potency for GPER binding would not be expected to allow GPER receptor identification using vacuum filtration methods related to its high off‐rates. Therefore to determine if aldosterone did bind to GPER, we studied its displacement of [3H] 2‐methoxyestradiol binding ([3H]2ME ‐ a higher affinity GPER‐selective agonist) in SF9 cells with heterologous GPER expression mediated by baculovirus.In this model system, following GPER gene transfer, [3H]2ME demonstrated saturable and reversible binding to a high affinity population of receptors with a Kd = 3.4±0.4 nM and Bmax = 645±49 fmol/mg. In the absence of baculovirus GPER gene transduction, no high affinity [3H]2ME binding was demonstrable in SF9 cells. [3H]2ME binding was guanine‐nucleotide sensitive with a 43±8% reduction in high affinity binding in the presence of GppNHp (100μM). High affinity [3H]2ME binding was displaceable by 2ME [with an IC50 comparable to the determined Kd (1.3±0.4 nM)], the GPER agonist G1, the GPER antagonist G15, estradiol (E2) and aldosterone (Aldo) but not by hydrocortisone (HC) with an order of potency of (2ME>Aldo>E2=G1>G15>>HC).In summary, we demonstrate high affinity [3H]2ME binding to GPER with an order of potency for aldosterone vs. estradiol (vs. hydrocortisone) that parallels the order of potency of their functional effects in previously studied models. These data support the hypothesis that aldosterone mediates its GPER‐dependent effects by direct binding to the receptor.Support or Funding InformationHeart and Stroke Foundation of CanadaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
15,16‐Dihydrotanshinone I, a natural product, protects ischemic stroke by inhibiting ferroptosis
Authors: Li Shaojing , Wu Chuanhong , Chen Xiuping 
Published: June 2021

Background/AimsIschemic stroke is a leading cause of death and disability worldwide. Early intervention of cell death in the penumbra can greatly reduce the infarct volume, thereby, alleviating the ischemic progress. Ferroptosis is a form of regulated cell death that is dependent on iron and reasctive oxygen species and is characterized by lipid peroxidation. Mitochondrial ROS are of importance in ferroptosis. Recent studies showed that it also implicated in the pathological cell death associated with stroke. Here, we studied the effect of 15,16‐Dihydrotanshinone I (DHT), a lipophilic tanshinone extracted from the root of Salvia miltiorrhiza Bunge (Danshen), on ischemic stroke and its underlying mechanism.MethodsPermanent middle cerebral artery occlusion (pMCAO)‐induced focal cerebral ischemia rats and tert‐butyl hydroperoxide (t‐BHP) injured PC12 cells were used to interpret the neuroprotective effect, especially the anti‐ferroptosis effect of DHT in vivo and in vitro.ResultsThe results showed that DHT protected t‐BHP injured PC12 cells and reversed ferroptosis induced by t‐BHP in vitro. It also protected rats subjected to pMCAO from ferroptosis in vivo. Furthermore, DHT ameliorated neurological score, infarct volume, cerebral blood flow, cerebral edema and microstructure of white‐grey matter in pMCAO rat in vivo.ConclusionThe data in this study demonstrated that DHT might have therapeutic potential for ischemic stroke and play its protective role against ferroptosis. This study reported that ferroptosis in the ischemic stroke represents a potential therapeutic target for the first time, and it might provide new insight into the prevention of ischemic stroke.Support or Funding Informationthe Science and Technology Development Fund of Macau Special Administrative Region (FDCT) (No. 078/2016/A2) , the Research Fund of University of Macau (MYRG2016‐00043‐ICMS‐QRCM), the National Natural Science Foundation of China (No. 81274133), the Major Scientific and Technological Special Project for “Significant New Drugs Creation” (No. 2018ZX09201008‐001, 2012ZX09103‐201‐055, ), the Fundamental Research Funds for the Central public welfare research institutes of China (ZZ2014060,ZXKT17040)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Antipsychotic Drug, Risperidone, Distributes to Bone Marrow: What is the Impact on Immune Function?
Authors: Houseknecht Karen L. , May Meghan , Beauchemin Megan , Barlow Deborah , Geguchadze Ramaz 
Published: June 2021

Antipsychotic medications are associated with serious side effects including obesity, dyslipidemia, diabetes, increased risk of fractures and bone loss. Additionally, it has been widely reported that schizophrenic patients are more susceptible to infections; however the role of antipsychotic medications (if any) in determining susceptibility to infectious disease is not understood. We have previously reported that risperidone (RIS) distributes to bone marrow of female mice following a single oral dose (Motyl et al., 2017, Bone 103:168–176).HypothesisThe bone marrow compartment may be a key target of risperidone action, resulting in altered immune function.MethodsTo test this hypothesis pre‐clinically, we treated male and female C57BL/6 mice with vehicle (VEH) or a low, clinically relevant dose of RIS (0.75 mg/kg, PO) for 5 days or 4 weeks. At the end of each treatment period, mice were sacrificed and plasma, bone marrow, and other tissues were collected for analysis. All animal study protocols were approved by the University of New England IACUC committee.ResultsConcentrations of drug and metabolites in plasma and bone marrow were determined by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) analysis. RIS and the active metabolite 9‐OH RIS (PAL) distributed to marrow of male mice following 5 days of dosing, and concentrations were higher in marrow than in plasma at 1 and 3 hours post‐dose (10–15 fold higher in marrow vs. plasma; P<0.05). Plasma concentrations of 40 cytokines or chemokines were measured in plasma from mice treated for 4 weeks with RIS or VEH using the Mouse Cytokine Array Panel (Proteome Profiler*TM Array, R&D Systems, Minneapolis, MN). Eight key immune mediators (IL‐17, INFγ, IL‐7, TIMP1, BCA‐1, IL‐13, IL‐1α, and I‐309) were significantly (P <0.5) downregulated in RIS‐treated mice. Hearts from VEH and RIS mice treated for 4 weeks were analyzed by RNASeq analysis and expression of 31 genes from immune/inflammatory pathways were significantly (2‐fold; P<0.05) altered (8 upregulated; 23 downregulated) in a manner consistent with global immunosuppression.ConclusionsThe highly prescribed antipsychotic, risperidone, distributes to bone marrow in mice and is associated with global immunosuppression as evidenced by reduced tissue (heart) and plasma concentrations of multiple cytokines and immune modulators. These data are consistent with increased susceptibility to infection, observed clinically. Future studies will explore the effect of RIS to modulate response to an immune challenge (in vivo).Support or Funding InformationFunding source: NIH grant DK095413 to KLHThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sodium propionate protects from inflammation and ROS stress
Authors: Lanza Marika , Casili Giovanna , Filippone Alessia , Campolo Michela , Paterniti Irene , Esposito Emanuela 
Published: June 2021

The major end‐products of dietary fibre fermentation by the gut microbiota are the short‐chain fatty acids (SCFAs) acetate, propionate and butyrate, which have been shown to modulate host metabolism via effects on metabolic pathways and receptor‐mediated mechanisms at different tissue sites. The beneficial effects of SCFAs on various aspects of gut physiology, barrier function, and metabolism have been well documented. Furthermore, SCFAs can promote intestinal homeostasis and suppress intestinal inflammation. Recently, several reports have been published describing inhibitory effects of SCFA on NF‐κB, one of the key transcription factors regulating genes implicated in innate immunity, cell cycle control, and apoptosis. However, most previous studies mainly focused on butyrate, and few studies have devoted their efforts to other SCFAs such as propionate, although it is abundant as butyrate in the gut and blood. Thus, the purpose of this study is to investigate protective mechanisms of sodium propionate (SP) in inflammatory and oxidative stress responses. The first step was to evaluate the anti‐inflammatory and ant‐oxidant mechanism of SP, by an in vitro model on murine macrophage cell line, J774‐A1, by two different approaches: stimulation with lipopolysaccharide (LPS) from E. coli and H2O2, both followed by the treatment with SP at different concentrations (0.1–1 and 10 mM ). Moreover, to evaluate the effect on acute inflammation, an in vivo model of Carrageenan (CAR)‐induced rat paw inflammation was performed, where rats were orally treated with different doses of SP (10 mg/kg, 30 mg/kg and 100 mg/kg). Our in vitro results showed that SP (0.1‐1‐10mM) significantly decreased in concentration‐dependent‐manner the expression of pro‐inflammatory mediators, such as cyclooxygenase‐2 (COX‐2) and inducible nitric oxide synthase (iNOS) following LPS stimulation. Moreover, it has been demonstrated that SP (0.1‐1‐10 mM) was able to enhance anti‐oxidant enzyme production such as manganese‐dependent superoxide dismutase (MnSOD) and heme oxygenase‐1 (HO‐1) following H2O2 stimulation. In addition, in in vivo model, SP(30 mg/kg and 100 mg/kg) markedly reduced paw inflammation, thermal hyperalgesia and tissue damage induced by CAR injection. Our results clearly demonstrated the anti‐inflammatory and anti‐oxidant properties of SP, therefore, we propose that SP may be an effective strategy for the treatment of inflammatory diseases.Support or Funding InformationNo fundingThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Matrix Metalloproteinase‐9 Release from Porphyromonas gingivalis Lipopolysaccharide (LPS)‐treated Rat Brain Microglia In Vitro
Authors: Memedovski Zylfi , Czerwonka Evan , Liao Shiting , Han Jin , Mayer Joshua , Hall Mary L , Mayer Alejandro M 
Published: June 2021

BackgroundThe putative association between periodontal disease (PD), a chronic inflammatory disease and neuroinflammation, a condition which involves brain microglia, remains under investigation. We have reported that Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS), a bacterium implicated in periodontitis, activates both classical (M1‐type) and alternative (M2‐type) activation of neonatal rat microglia (BMG) with concomitant release of superoxide anion, thromboxane B2, cytokines, and chemokines (Faseb Journal 32.1 Supplement: 702.2, 2018). We hypothesized that matrix metalloproteinase 9 (MMP‐9) release by Pg‐treated microglia might be detectable by both ELISA and visualized by confocal fluorescence imaging.MethodsP. gingivalis LPS (Pg LPS) 105 ng/mL from InvivoGen (San Diego, CA) and Escherichia coli LPS (Ec LPS) 026:B6 1 ng/mL from Difco Lab, Detroit, MI (positive control) were used to treat rat BMG for 18 hours at 35.9°C in vitro. MMP‐9 release was assessed with a rat MMP‐9 ELISA kit (R&D Systems). Confocal fluorescence imaging with a Nikon A1R laser confocal microscope was used to identify BMGs using a primary mouse anti‐rat CD11b/c antibody (Ab) (AbD SeroTec, Raleigh, NC), and secondary donkey anti‐mouse Ab (Thermo Fischer, Waltman, MA). MMP‐9 was visualized with a primary rabbit anti‐rat MMP‐9 polyclonal Ab, and secondary goat anti‐rabbit Ab, both from Abcam, Cambridge, MA.ResultsMMP‐9 presence in tissue culture supernates was detectable by ELISA in both Ec LPS and Pg LPS‐treated BMG. In contrast, MMP‐9 was observed by confocal fluorescence imaging only within Ec LPS‐treated BMG but not in Pg LPS‐treated BMG.ConclusionsBoth ELISA and confocal fluorescence imaging provide additional support for our working hypothesis, because Pg LPS stimulated BMGs to release MMP‐9. Thus, our current observations extend our studies on the effect of Pg LPS on rat brain microglia activation and contribute to further characterize the putative role of Pg LPS in both periodontitis, neuroinflammation and neurodegeneration.Support or Funding InformationSupport by College of Dental Medicine, College of Graduate Studies, and Chicago College of Osteopathic Medicine, 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.
Interaction of Local Anesthetics with Hepatocellular Organic Anion Transporting Polypeptides
Authors: Stieger Bruno , Antener Enrica , Häusler Stephanie , Kullak‐Ublick Gerd A. 
Published: June 2021

Organic anion transporting polypeptides (OATPs) have a very broad substrate specificity and are key transporters involved in drug disposition. Some OATPs display a broad tissue distribution. The substrates of OATPs are often anions, but also include neutral and cationic compounds and can be poorly water soluble. Local anesthetics can be divided chemically into amino esters and amino amides. Local anesthetics may undergo hepatic elimination and therefore require transport systems for hepatic uptake prior to metabolism. The aim of this project therefore is to test for a possible interaction of local anesthetics with the hepatocellular OATPs: OATP1B1, OATP1B3 and OATP2B1. Cells stably expressing hepatocellular OATPs (DMD 35: 1400, 2007; EJP 584: 57, 2008) were used for transport experiments. Inhibition of local anesthetics on the transport of standard OATP substrates was investigated as previously described (DMD 35: 1400, 2007; EJP 584: 57, 2008). So far, the two prototypic local anesthetics lidocaine (amine ester) and procainamide (amine amide) were tested using this approach. We found no interaction of these two local anesthetics on transport mediated by either of the three OATPs tested. We conclude that testing additional local anesthetics is necessary for testing our hypothesis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.