bluuebunny/crossref_metadata_2025_split huggingface dataset:
Phosphatidylethanolamine‐Binding Protein Promotes Opioid Anti‐Nociception in the Brain and Spinal Cord by Reducing βarrestin2 Recruitment to the Mu Opioid Receptor
Authors: Kim Caleb Ya Ill , LaVigne Justin , Edwards Katie , Streicher John M. 
Published: June 2021

Opioids have been widely accepted as an effective treatment for chronic pain at a cost of serious side effects, including tolerance and addiction. Side effects are caused in part by desensitization and downregulation of mu opioid receptors (MOR) through recruitment of βarrestin2. Phosphatidylethanolamine‐Binding Protein (PEBP) is known to regulate either Raf Kinase or G Protein Receptor Kinase‐2 (GRK2) through phosphorylation by Protein Kinase C (PKC), and GRK2 is a major player in the recruitment of βarrestin2. However, PEBP has not been investigated for its role in MOR signaling and opioid response. We first tested this using an in vitro model of βarrestin2 recruitment in MOR expressing U2OS cells. We found that by inhibiting PEBP through siRNA knockdown or the small molecule inhibitor locostatin, we could increase the recruitment of βarrestin2 to the MOR in 2 separate assays. Conversely, we found that activating PKC via phorbol myristate acetate (PMA) led to PEBP phosphorylation/activation, and a reduction in βarrestin2 recruitment to the MOR, which could be blocked by PEBP knockdown. We then tested the role of PEBP in vivo by injection of locostatin into the brain or spinal cord, or CRISPR editing of PEBP in the spinal cord. We found that blocking PEBP in vivo led to a strong reduction in morphine‐induced tail flick anti‐nociception in both the brain and spinal cord. Together, these results support the hypothesis that PEBP acts to sequester GRK2 and block βarrestin2 recruitment downstream of the MOR both in vivo and in vitro. These results also suggest that enhanced PKC/PEBP activation by opioid drugs could further reduce opioid side effects and enhance analgesic efficacy.Support or Funding InformationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Reactive Skin Decontamination Lotion (RSDL) Maintains Its Effectiveness against Cutaneous Applications of VX Nerve Agent for at Least 5 Years Past Its Expiration Date
Authors: Clarkson Edward D. , Morgan Julia E. , Chen Paul , Schultz Susan M. , Soni Sunil D. , Capacio Benedict R. , Koplovitz Irwin 
Published: June 2021

Reactive Skin Decontamination Lotion (RSDL) is the primary product the US military has fielded to decontaminate cutaneous exposures to chemical warfare agents. RSDL is extremely effective at protecting guinea pigs against cutaneous applications of the nerve agent VX. The 24‐hour cutaneous LD50 of VX in un‐anesthetized haired clipped guinea pigs is 140 µg/kg. When RSDL is used to remove VX two minutes after application the LD50 is 7,310 µg/kg. But RSDL is expensive to field and has a limited operational life. In the summer of 2017 we addressed the question of reducing costs by comparing the effectiveness of two expired (2012, 2014) lots of RSDL, to a lot that expires in April 2020. We applied the LD30 dose of 5,500 µg/kg to groups of n=12 and used the different batches of RSDL to remove the VX. There was no statistical difference in survival (n=10, 8 and 9) between the groups. The RSDL packets did change color as they aged, with the non‐expired RSDL having a bright yellow color, the RSDL that expired in January 2014 having a gray color and the RSDL that expired in April 2012 having a black color. Analysis of the various RSDL packets by proton, carbon and phosphorus NMR spectroscopy indicated that the difference in the component composition was not greater than 1%. To determine if protection was due to physical removal of VX we exposed two groups of n=4 to 5,500 µg/kg and 2,250 µg/kg of VX and used RSDL trainers to remove the VX. In the 5,500 µg/kg group all 4 animals died, and in the 2,250 µg/kg group 2 animals died. These results argue that RSDL can be effective past its expiration date and that the protection provided is not due entirely to physical removal.The views expressed in this abstract are those of the author(s) and do not reflect the official policy of the Department of Army, Department of Defense, or the U.S. Government. The experimental protocol was approved by the Animal Care and Use Committee at the United States Army Medical Research Institute of Chemical Defense and all procedures were conducted in accordance with the principles stated in the Guide for the Care and Use of Laboratory Animals and the Animal Welfare Act of 1966 (P.L. 89‐544), as amended.Support or Funding InformationThis research was supported by the Defense Threat Reduction Agency – Joint Science and Technology Office, Medical S&T Division.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Quantitative Assessment of Trimethyltin, Kainic Acid and Pyrithiamine Neurotoxicity in Rat Brain and its Correlation with MR Imaging
Authors: Srivastava Anshul , Liachenko Serguei , Paule Merle G. , Sarkar Sumit , Pandey Jai , Negi Geeta , Hanig Joseph 
Published: June 2021

Toxicagents such as trimethlytin (TMT), kainic acid (KA) and pyrithiamine (PT) induce neurotoxicity. Previous studies have reported degeneration in cortex, hippocampus, olfactory bulb, thalamus a swell as various other brain areas following exposure. Quantitative assessment of the extent of the degeneration is essential for understanding the functional changes due to such toxicity. In the current study, quantitative analyses of the pathological changes associated with single i.p. injections of TMT(12 mg/kg), KA (10 mg/kg) and PT (.25 mg/kg) were performed using tissue classification and area quantification algorithms based on stain color and OD available in HALO™ image analysis software. Sections containing brain areas to be compared to MRI images were chosen from 81 coronal sections of the whole rat brain stained with amino‐cupric silver and subsequently digitalized. Rats treated with TMT showed degeneration in amygdaloid nucleus (11.79%), cortex (7.71%), hippocampus(6.28%), hypothalamus (5.81%), septal region(4.92%), olfactory bulb (4.72%), thalamus (2.82%), nucleus accumbens (2.34%), caudateputamen (2.11%) and substantia nigra (0.40%). Cortical and hippocampaldegeneration in KA treated animals were 5.78% and 4.47%, respectively. Thalamusand olfactory degeneration in PT treated rats were 3.99% and 1.56% respectively. Saline treated animals served as controls and showed <0.3% changes in these brain areas. Quantification results of histopathology using image analysis were supported by the pathology score where the affected area was scored based on a severity scale. MRI T2 responses in hippocampus and cortex were compared to the toxicity in TMT treated animals as determined via histopathology quantification. Receiver Operating Characteristic (ROC) curves were generated to correlate histopathology and MRI responses. Correlation of MR imaging with quantitative histopathology analyses showed that MRI signals were in concordance with the histopathology and could effectively estimate loci and extent of neurodegeneration. Results suggest that MRI can be used as non‐invasive biomarker for the detection of drug‐induced neuropathology.Support or Funding InformationCenter for Drug Evaluation and Research ‐ Critical Path Award and National Center for Toxicological Research, Food and Drug AdministrationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interaction of Respirable Fracking Sand Dust (FSD) with Pulmonary Tissues In vivo and In vitro
Authors: Fedan Jeffrey S. , Thompson Janet A. , Russ Kristen A. , Roberts Jenny R. , Schwegler‐Berry Diane , Mercer Robert R. 
Published: June 2021

Sand added to fracking fluid that is pumped into the well bore is used as a proppant to maintain fissures in fractured shale to facilitate gas flow during unconventional gas drilling. Mechanical manipulation of the sand during preparation of fracking fluid gives rise to respirable dust (fracking sand dust; FSD) and inhalation exposures of workers to crystalline silica at levels in excess of regulatory standards. We characterized properties of FSD and some of its biological effects. Analysis of worksite‐derived FSD particles (≤2 μm major axis; >95% crystalline silica) usingenergy‐dispersive X‐ray spectroscopy (EDS) revealed eight categories of differing compositions based on the presence of Si, O, Al, Mg, K, Ca, Mn, Fe, and Co; elemental heterogeneity was present in individual particles. Inhalation of FSD by rats (6 h/d, 4 d, 10 or 30 mg/m3) led to dose‐dependent lung burdens at 1, 7, and 27 dpost‐exposure, with clearance noted following 1 d post‐exposure. In enhanced dark field (EDF) images of trachea, particles associated with epithelium were observed 1 d but not 7 or 27d post‐exposure. EDF studies of clearance revealed that particles in the alveolar region of the lungs were reduced to ~50% and ~30% by 7 and 27 d post‐exposure; none were present at 90 dpost‐exposure. Clearance of FSD appears to be more rapid than that reported for pure crystalline silica (i.e., MIN‐U‐SIL). Previously, inhalation of 30 mg/m3 FSD was observed to inhibit tracheal epithelial active Na+ transport, suggesting that the epithelium is a target of FSD effects. We, therefore, examined the effects of 18 hincubation with apical FSD (0.0001 – 1 mg) on cytokine and lactatedehydrogenase (LDH) release into apical and basolateral media from normal humanbronchial epithelium in air‐interface culture. Changes in cytokine levels in a 65‐cytokine panel of growth factors and inflammatory mediators were modest (0.1 – 1 mg). No LDH was released into the basolateral medium; LDH released into the apical medium was unaffected by FSD. The results indicate that FSD is fairly rapidly cleared from the lungs following inhalation and is without appreciable direct toxicity to airway epithelial cells.Support or Funding InformationNIOSH 8927ZLDCThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Graphene interactions with human endothelium
Authors: Szukiewicz Dariusz , Skoda Marta , Dudek Ilona 
Published: June 2021

The discovery of graphene boosted an interdisciplinary research based on its unique properties and possible applications in biomedicine. Multiple scientific reports concerning graphene interactions with adherent cells showed the potential use of graphene as a biomaterial in tissue engineering.In this study we examined the influence of pristine graphene, as a monolayer coating or flake suspension, on human endothelial cells (HUVEC) isolated from umbilical veins. Umbilical cords were obtained from healthy gravidas after the cesarean section procedures. Cytotoxicity, proliferation and cell morphology study were performed using various methods, including colorimetric and luminescent cell assays, optical and fluorescence microscopy.The cytotoxicity assay confirmed dose and time dependent cytotoxicity of tested graphene flakes (G1APS~5000nm and G2APS~450nm). Even the lowest concentration of graphene (10μg/ml) significantly inhibited cell proliferation. The difference between the two tested flake sizes was more distinct during cell mortality study, indicating the greater cytotoxic effect of the smaller graphene particles. Graphene suspended in the cell culture medium created aggregates. Cell imaging revealed efficient accumulation of nanoflakes inside the cells, especially those of the smaller size.The morphology of the cells seeded on graphene‐coated SiO2 substrates was recorded using fluorescence microscopy. Endothelial cells were immunostained for actin cytoskeleton to characterize its organization, stress fibers creation, cell adhesion and spreading. After a 72h incubation, both the cells incubated on graphene SiO2 substrates and on the reference material for cell plating, reached confluence. No difference in cell shape and morphology was detected. Graphene was shown to be a suitable surface for endothelial cell growth.Our preliminary investigation confirmed cytotoxic effect of pristine graphene flakes on adherent cells, however demonstrated that CVD grown graphene is not toxic for human endothelial cells. These data show promise for using graphene monolayers in tissue engineering or as a substrate material for implant devices.Support or Funding InformationWUM grant: 2M2‐W2‐16This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
atRA induces cholesterol accumulation through CYP7A1 repression in the liver
Authors: Won Kyoung‐Jae , Jeong Hyunyoung 
Published: June 2021

All‐trans retinoic acid (atRA) is one of the most frequently used retinoids to treat certain cancers or dermatological diseases. Common side effects of atRA include hypercholesterolemia, leading to discontinuation of atRA uses. Previous studies have shown that atRA downregulates CYP7A1 (the rate‐liming enzyme converting cholesterol to a bile acid intermediate), potentially responsible for hypercholesterolemia. However, the detailed molecular mechanisms in atRA‐induced hypercholesterolemia is unclear. atRA increased cholesterol levels in a dose dependent manner in HepaRG cells, and this was accompanied by dramatic decrease in CYP7A1 mRNA and protein levels. CYP7A1 overexpression by lentiviral system reversed atRA‐induced cholesterol accumulation in HepaRG cells, suggesting that the cholesterol accumulation is mediated by CYP7A1 repression. CYP7A1 promoter reporter assays in HepG2 cells revealed that the HNF4α binding site in the promoter (−149/−118) is essential for atRA‐mediated CYP7A1 repression. HNF4α depletion by siRNA abrogated CYP7A1 repression by atRA in HepaRG cells. Western blot analysis revealed that atRA activates MAPKs signaling pathways including JNK, ERK, and p38 MAPKs in HepaRG cells. Pharmacological inhibition of JNK and ERK pathways but not p38 pathway by using a specific inhibitor (i.e., SP600125, PD98059, and SB203580 for JNK, ERK, and p38, respectively) attenuated atRA‐mediated CYP7A1 repression and cholesterol accumulation. Overexpression of AP‐1 (c‐Jun/c‐Fos), a downstream target of JNK and ERK, repressed CYP7A1 expression in HepG2 cells. In DNA pull‐down assay, AP‐1 exhibited sequence‐specific binding to the HNF4α binding site in CYP7A1 promoter following atRA treatment in HepaRG cells. Chromatin immunoprecipitation assay also showed that AP‐1 binding to the CYP7A1 promoter (−181/−34) was increased following atRA treatment, whereas HNF4α binding was decreased. Collectively, results from this study indicate that atRA‐activated JNK and ERK pathways and downstream target AP‐1 represses HNF4α transactivation of CYP7A1 promoter. These results suggest an important role of CYP7A1 in atRA‐induced cholesterol accumulation and provide a basis to establish future strategy in cholesterol disorder by atRA.Support or Funding InformationThis study was supported by R01 HD089455.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Suppression of Polycyclic Aromatic Hydrocarbon (PAH)‐Mediated Pulmonary Carcinogenesis in Mice by Omega‐3‐Fatty acids
Authors: Moorthy Bhagavatula , Jiang Weiwu , Wang Lihua , Maturu Paramahamsa , Zhou Guodong 
Published: June 2021

3‐Methylcholanthrene (MC) and benzo[a]pyrene (BP) are potent polycyclic aromatic hydrocarbons (PAHs) carcinogens. PAHs are present in cigarette smoke, diesel exhausts, and charcoal broiled meats, etc. Cytochrome P450 (CYP) 1A/1B1 enzymes play key roles in the activation of PAHs to carcinogenic metabolites, which initiate carcinogenesis by binding covalently to DNA, and the adducts, if not repaired, could lead to tumorigenesis. In this study we tested the hypothesis that pre‐treatment of mice with omega‐3‐fatty acids, i.e. [eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) will lead to attenuation of PAH‐mediated pulmonary DNA adduct formation and pulmonary tumorigenesis. Twelve week old male and female A/J mice received EPA (60 mg/kg) and DHA (40 mg/kg) from day 1 to day 24. Control mice were treated with vehicle corn oil. On day 3, mice were treated with BP (40 μmol/kg) or 3‐methylcholanthrene (MC, 40 μmol/kg) by i.p. In the short‐term experiment (DNA adduct studies), 3–5 mice from each group were terminated at day 10 (7 days after BP or MC treatment). EPA/DHA significantly suppressed formation of BP‐DNA and MC‐DNA adducts in lung and liver of both male and female mice. In the long‐term experiment (tumor studies), 8 mice for each group continued to receive EPA/DHA twice per week till end of experiment (16 weeks). Pulmonary tumor incidence and multiplicity was significantly suppressed in mice given EPA+DHA, compared to those given the PAHs, BP or MC alone. PAH treatment led to enhancement of expression of epigenetic genes such as DNMT and EZH2 and repression of RUNX3 in lung. Mice given a combination of PAHs and EPA;/DHA attenuated DNMT and EZH2, but augmented RUNX3. These results suggest that EPA/DHA contribute to the suppression of PAH‐mediated mechanisms in part by modulating epigenetic genes. These studies also suggest that the omega 3‐fatty acids could be developed as novel candidates for the prevention of PAH‐mediated lung cancers in humans.Support or Funding InformationR01ES 009132 and R01HL129794This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Obesity Alters the Expression of Suppressive Factors in Intra‐Tumoral Dendritic Cells Following Combinatorial Immunotherapy
Authors: Turbitt William , Boi Shannon , Orlandella Rachael , Gibson Justin , Norian Lyse 
Published: June 2021

A growing literature suggests that obesity induces dysregulated antitumor immunity. We found previously that diet‐induced obesity (DIO) increases the abundance of intra‐tumoral regulatory dendritic cells (DCs) in a murine model of renal cancer. Furthermore, DIO reduces the efficacy of an immunotherapy consisting of intra‐tumoral delivery of replication‐deficient adenovirus (Ad) encoding tumor necrosis factor‐related apoptosis‐inducing ligand (TRAIL) (AdT) plus class B oligonucleotides (CpG1826). This therapy primes endogenous T cells by targeting DCs that process and present tumor‐derived antigens released by TRAIL‐mediated tumor cell death. Our current study evaluated the efficacy of AdT/CpG combined with either anti‐PD‐1 or anti‐CTLA‐4 immune checkpoint blockade, in the absence or presence of DIO. BALB/c mice (7–8‐weekold) were randomized to standard chow or high‐fat diet for 20 weeks prior to the orthotopic injection of Renca‐Luc renal cancer cells into the left kidney and were maintained on diets until day 28 post‐tumor injection. At day seven post‐tumor injection, mice were re‐injected in the left kidney with saline or AdT/CpG. Mice were further randomized to receive intra‐peritoneal isotype control, anti‐PD‐1, or anti‐CTLA‐4. Both AdT/CpG+anti‐PD‐1 (p<0.001) and AdT/CpG+anti‐CTLA‐4 (p<0.001) resulted in a significant reduction in renal tumor outgrowth and spontaneous lung metastasis in lean mice on standard chow. AdT/CpG increased the accumulation of intra‐tumoral DCs independent of dietary or immunotherapy intervention, with the conventional DC two (cDC2) subset favored over the conventional DC one (cDC1) subset. Expression of PD‐1 and PD‐L1 on DC subsets was unchanged in response to diet or immunotherapy intervention; however, untreated tumor‐bearing DIO mice had elevated PD‐1 expression on cDC1s (p=0.022) compared to untreated lean mice or lean mice receiving AdT/CpG+anti‐PD‐1. Lastly, Arginase‐1 expression in cDC1s was not significantly different between groups; however, Arginase‐1 expression was elevated in response to DIO on cDC2s (p=0.006) compared to both lean and DIO AdT/CpG+anti‐PD‐1 treated mice. Future studies are needed to characterize the suppressive function of intra‐tumoral DC populations, as well as the T cell priming capacity of DCs within tumor‐draining lymph nodes in response to immunotherapy in the absence or presence of DIO. In conclusion, the differential response to immunotherapy and alteration in suppressive factors in intra‐tumoral DCs in DIO mice warrants further investigation, as targeting these pathways may ultimately enhance treatment strategies in overweight and obese cancer patients.Support or Funding InformationRO1CA181088 and T32DK062710This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Central Cholinergic Pathways Diminish the Hypotensive and Cardiac Autonomic Depressant Effects of Endotoxemia in Male Rats: Role of Medullary NFκB/α7/α4β2 nAChR Signaling
Authors: Sallam Marwa M. , El‐Gowilly Sahar M. , Fouda Mohamed A. , Abd‐Alhaseeb Mohamed M. , El‐Mas Mahmoud M. 
Published: June 2021

We have recently shown that systemically administered nicotine improves endotoxic manifestations of hypotension and cardiac autonomic dysfunction in rats. In this report, integrative and molecular studies were employed to investigate whether medullary antiinflammatory pathways of α7 and α4β2 nicotinic acetylcholine receptors (nAChRs) modulate cardiovascular derangements induced by endotoxemia. The i.v. administration of nicotine (50 mg/kg) to conscious rats abolished the lipopolysaccharide (LPS, 10 mg/kg i.v.)‐evoked: (i) falls in blood pressure, (ii) decreases in spectral measure of cardiac sympathovagal balance (ratio of the low‐frequency to high‐frequency power, LF/HF) and (iii) increases in the immunohistochemical protein expression of NFκB in medullary neurons of the nucleus tractus solitarius (NTS). These favorable influences of nicotine were replicated in rats treated intracisternally (i.c.) with PHA‐543613 (selective α7 nAChR agonist; 2 μg/rat), or 5‐iodo‐A‐85380 (5IA, selective α4β2 nAChRs agonist; 0.1 μg/rat). Measurement of arterial baroreflex activity by the vasoactive method revealed that nicotine, PHA, or 5IA reversed the depressant effect of LPS on reflex bradycardic, but not tachycardic, activity. Pharmacologic antagonist studies showed that the reversing effect of nicotine on LPS hypotension was partly inhibited in rats treated concomitantly with i.c. methyllycaconitine citrate salt hydrate (α7 nAChR antagonist, 10 μg/rat) or dihydro‐β‐erythroidine hydrobromide (α4β2 nAChR antagonist, 10 μg/rat), whereas the associated increases in LF/HF ratio remained unaltered. The data signifies the importance of the NTS inflammatory pathways in cardiovascular endotoxic manifestations and the capacity of brainstem circuits of α7 and α4β2 nAChRs in ameliorating these responses.Support or Funding InformationSupported by the Science and Technology Development Fund, Egypt (STDF Grant No. 14895)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.