bluuebunny/crossref_metadata_2025_split huggingface dataset:
Proteomic Changes After Surgical Repair in Congenital Heart Diseases
Authors: He Guo‐Wei , Hou Hai‐Tao , Xuan Chao , Wang Jun , Liu Li‐Xin , Zhang Jian‐Feng , Yang Qin 
Published: June 2021

BackgroundThe final goal for treatment of congenital heart diseases (CHD) is to resume not only the normal heart structure but also physiology. The present study evaluates the proteomics changes before and after surgery in CHD with tetralogy of Fallot (TOF) or ventricular septal defect (VSD).MethodsDifferential plasma proteins by using two‐dimensional gel electrophoresis and mass spectrometry followed by ELISA validation in new cohort were analyzed in patients with TOF (n=82) or VSD (n=82) compared to normal controls (n=97) preoperatively and 6‐month postoperatively.ResultsA total of 473 protein spots preoperatively and 515 postoperatively including those significantly (p<0.01) downregulated or upregulated were identified. The ELISA validation for chosen proteins demonstrated that in VSD patients, postoperative alpha‐1‐acid glycoprotein 2 and serum amyloid p‐component were completely recovered (p>0.05, compared to control) and complement component C3c was partially recovered (p<0.05, compared to control) towards normal after surgery. In TOF patients, postoperative fibrinogen gamma chain was completely and gelsolin was partially recovered.ConclusionsWe have for the first time demonstrated that CHD repair surgery not only corrects the structure malformation but also resumes the normality of altered proteins that may be developed as biomarkers. Identification of the recovered or unchanged proteins may be used for evaluation of the surgical results and for facilitating the personalized management for CHD.Support or Funding InformationSupported by grants from the National Natural Science Foundation of China (81170148 & 81641017), Zhejiang Provincial Natural Science Foundation (LY15H020008), & Tianjin Binhai Key Platform for Creative Research Program (2012‐BK110004), Binhai New Area Health Bureau (2012BWKZ008, 2016BWKY007, 2016BWKZ003).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 relationship between anemia and sudden cardiac death in severe aortic stenosis
Authors: Ducharme‐Smith Allison , Chahal Anwar A. , Gersh Bernard J. , Somers Virend K. , Sawatari Hiro , Brady Peter A. , Nkomo Vuyi T. , Pellikka Patricia A. 
Published: June 2021

BackgroundAortic stenosis (AS) affects, and is associated with significant comorbidity and mortality, including sudden cardiac death (SCD) from unknown mechanisms. However, it is difficult to predict which patients with AS will suffer a SCD event. Anemia and low iron are known risk factors for mortality in heart failure populations and AS. We hypothesized that anemia in patients with AS is associated with increased risk of SCD.PurposeUnderstand the relationship and prognostic implications between anemia and SCD in patients with severe AS.Methods and ResultsWe selected 8,144 of 10,850 patients with severe AS (mean transvalvular gradient ≥40 mmHg or peak velocity ≥4 m/s or calculated valve area < 1 cm2) from the Mayo Clinic Aortic Stenosis Registry, which includes all consecutive patients with severe AS over the time period January 1st, 1983 to September 30th, 2015. Subjects in this analysis (median age 76 years and 57% male) were diagnosed starting in 1995 and had hemoglobin (Hb) measurements within one year of diagnosis and prior to intervention. The mean Hb level was 12.8 (± 1.9) g/dL. The prevalence of anemia (Hb<13.0g/dL for men, <12.0g/dL for women) was 40%, 14% microcytic, 20% macrocytic, with the remainder normocytic. Of the 8,144 patients, a total of 4,440 died (10‐year survival 39%; and 240 were confirmed as SCD events [10 year cumulative incidence 4%]). Anemia was associated with increased all‐cause mortality (HR 1.87, 95% CI 1.76–1.99, P=<0.001) and increased SCD events (HR 1.58, 95% CI 1.22–2.06, P=<0.001) in patients with severe AS. Every SD decrease in Hb (1.9 g/dL) was associated with an increased risk of SCD (HR 1.42, 95% CI 1.24–1.62, P<0.001).ConclusionsAnemia is highly prevalent in patients with severe aortic stenosis and is independently associated with increased all‐cause mortality, and more specifically with increased SCD risk. Possible mechanisms include anemia aggrevating myocardial hypoxia, increasing propensity for arrhythmia and abnormal Bezold‐Jarisch reflex mismatch. Anemia may be a useful prognostic marker and a potential therapeutic target in patients with severe AS.Support or Funding InformationCAAC is supported by AHA17POST33400211 and NIH HL134885. This study was supported by a grant from the Department of Cardiovascular Medicine, Mayo Clinic Rochester.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A comparative study on phenotypic and molecular characteristics of ischemic stroke resistant animal model
Authors: Kang hyunbon , Won Jinyoung , Hong Yonggeun 
Published: June 2021

The use of animal stroke models has substantially increased to develop the therapeutic agents in preclinical studies according to the high mortality rate of cardiovascular disease. Animal ischemic stroke model is generally classified into three types, global ischemia, focal ischemia, and hypoxic ischemia. These animal models have been extensively used in ischemic stroke studies. However, ischemia induced injury models reveal several limitations to identify underlying pathogenesis of stroke. The purpose of this study was to develop the stroke resistant model using ischemia induced cell death signal targeted gene, DAPK1/Pin1 deletion. In addition, stroke resistant model was screened the major phenotypes, physiological functions, and molecular characteristics in organ system. DAPK1/Pin1 single gene knockout (KO) and double gene KO (DKO) mice were generated to determine whether DAPK1 and Pin1 contribute to neuronal death in ischemic injury. The genotype was confirmed by PCR using genomic DNA. DAPK1/Pin1 DKO mice were produced by crossing female DAPK1/Pin1 heterozygous (hetero) with identical genotype male mice. The offspring from DAPK1/Pin1 hetero mice showed significant difference compared with wild type (WT). Body weight and body length of Pin1 KO and DAPK1/Pin1 DKO mice were less than WT and DAPK1 KO mice. DAPK1/Pin1 DKO mice exhibited restricted growth rate. To examine the therapeutic effects of DAPK1/Pin1 deficiency, KO mice were subjected to MCAo. DAPK1/Pin1 DKO mice exhibited smaller infarct size and reduced neurologic deficits compared with single gene KO. Taken together, ischemic stroke resistant model induced by DAPK1/Pin1 genetic suppression could be potential targets for treating stroke.Support or Funding InformationFunds: NRF‐2013R1A2A2A01067169, NRF‐2017R1A2A2A01067169, KRIBB‐KGM4611714This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CXCR4 inhibitor, MSX‐122 suppresses AOM‐induced colon cancer in Apc+/Min mouse
Authors: Bissonnette Bruce Marc , Dougherty Urszula , Mustafi Reba , Haider Haider I. , Joseph Loren , Souris Jeffrey , Hart John A. , Pewkow Joel R. , LI Yan Chun 
Published: June 2021

IntroductionIn prior studies, we showed that EGFR is required for tumor promotion by Western diet in the azoxymethane (AOM) model of sporadic colon cancer and the AOM/DSS model of inflammation‐associated colon cancer. We also demonstrated that ADAM17, the metalloprotease responsible for shedding membrane‐bound EGFR ligands, controls tumor development in both the AOM and the Apc+/mutant multiple intestinal neoplasia (Min) mouse model. Furthermore, we established that Sdf1a‐CXCR4 signals activated EGFR by an ADAM17‐dependent mechanism in colon cancer cells; and that Western diet up‐regulated Sdf1a‐CXCR4 and ADAM17 in colonic mucosa. We hypothesized that CXCR4 blockade might inhibit colon cancer development. The aim of this study was to test the ability of a novel CXCR4 antagonist, MSX‐122 (substituted benzylamine) to inhibit AOM‐promoted colon cancer in Apc+/Min mice. Min mice without AOM treatment mostly develop small intestinal tumors, whereas AOM‐treated Apc+/Min mice develop colon cancers (Suzui et al 2002).MethodsIn studies in cell culture, we examined the ability of MSX‐122 to block Sdf1a‐induced EGFR transactivation. In vivo, Apc+/Min mice were given AOM (10 mg i.p./kg body weight) and two weeks later mice were started on a Western diet (20% fat). Mice received MSX‐122 i.p. (2.5 mg/kg body weight, n=12) or vehicle (DMSO, n=13) three times weekly and were killed after 10 wks.ResultsMSX‐122 blocked EGFR transactivation by Sdf1a in HT29 colon cancer cells as assessed by reductions in pErbB2, pAKT and pERK. In vivo, MSX‐122 was well tolerated with mice showing comparable weight gain to vehicle‐treated mice. As expected, AOM induced colon tumors in Apc+/Min mice. MSX‐122 significantly reduced colon tumor incidence (92.3% vs. 50.0%, p=0.025) and cancer incidence (69.2% vs. 16.7%, p=0.015). MSX‐122 also reduced tumor burden as assessed by total tumor multiplicity (1.8 vs. 1.0 tumor per TBM; p=0.02), tumor volume (169.5 mm3 vs. 61.2 mm3, p<0.023) and tumor proliferation as assessed by Ki67 staining (37.2±3.6% vs. 25.3±5.8%, p<0.005). Cancer multiplicity also appeared to be decreased (1.6 vs. 1.0 tumor/TBM, p=0.17).ConclusionsMSX‐122 blocked EGFR transactivation by CXCR4 agonist Sdf1a in colon cancer cells. MSX‐122 also inhibited colon tumor development and progression in the AOM‐promoted Apc+/Min mouse model. We speculate that MSX‐122, which was well tolerated in a phase 1b clinical trial (Painter, W. 2015), could serve as a chemopreventive agent in individuals at increased risk for colon cancer.Support or Funding InformationNIH R01CA164124This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cannabinoid Type‐1 Receptors can Mediate the Antinociceptive Effects of Heroin in Nonhuman Primates
Authors: Nilges Mark R. , Bondy Zachary , Grace Joseph Allie , Winsauer Peter 
Published: June 2021

Opioid analgesics can provide adequate pain relief under some conditions; however, adverse effects such as dependence, abuse liability, and respiratory depression are major clinical complaints. Cannabinoids such as THC (the psychoactive constituent in marijuana) can produce analgesic effects under some conditions, but generally at higher doses that produce behaviorally disruptive effects. Combinations of opioids and THC produce thermal antinociceptive effects in a synergistic manner, such that ineffective doses of each drug can produce antinociception when given in combination. In this experiment, male nonhuman primates (rhesus macaques) were administered THC in combination with cumulative doses of heroin to determine if THC enhances the antinociceptive effects of heroin. Furthermore, we tested the effects of rimonabant, a selective cannabinoid type‐1 receptor (CB1R) antagonist, to determine the antinociceptive contribution of CB1R. Antinociception was assessed using a tail‐dip procedure. Briefly, the animals were secured in standard primate chairs, tails were shaved, and dipped in a mixed order in three different water temperatures (40°, 50°, and 55°C) until the tail was withdrawn or 20 seconds elapsed. Latency (sec) to remove the tail was converted to percent maximum effect (%MPE). The initial tail dips of a session served as the baseline latency for each temperature, and the subsequent injections were either vehicle (control), THC (0.1 – 1 mg/kg), and/or rimonabant (0.32 mg/kg). These injections were then followed by cumulative doses of either heroin (0.032 – 0.32 mg/kg) or vehicle. Each set of tail dips was separated by a 15‐min period that also served as an inter‐injection interval for the cumulative doses of heroin. When THC was administered prior to heroin, there was a dose‐dependent leftward shift in the heroin dose‐effect curve at both 50° and 55°C temperatures. When rimonabant (0.32 mg/kg) was administered prior to heroin, there was a rightward shift in the heroin dose‐effect curve at the 50°C water temperature, but no change was detected at 55°C. When rimonabant (0.32 mg/kg) preceded THC (1 mg/kg), the enhancement of heroin‐induced antinociception was partially antagonized at the 50°C water temperature, but not at 55°C. Overall, rimonabant attenuated the antinociceptive effects of heroin alone and partially antagonized the enhanced antinociceptive effects of the THC‐heroin combinations at the lower stimulus temperature. These data are consistent with the notion that THC can potentiate the antinociceptive effects of an opioid agonist in a CB1‐dependent manner.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Biophysical identification of the 5‐HT2A receptor:5‐HT2C receptor interaction in vitro
Authors: Felsing Daniel Eric , Anastasio Noelle C. , Miszkiel Joanna M. , Gilbertson Scott , Moeller Frederick Gerrard , Allen John A. , Cunningham Kathryn A. 
Published: June 2021

Disruption of serotonin 5‐HT2A receptor (5‐HT2AR) and 5‐HT2CR function is implicated in a range of neuropsychiatric disorders with these receptors being essential sites of action for anti‐psychotic and anti‐obesity medications. Based on evidence that the 5‐HT2AR and 5‐HT2CR co‐localize and co‐immunoprecipitate from neurons in the medial prefrontal cortex of the rat (Anastasio et al, ACS Chem. Neurosci. 15:1248, 2015), we hypothesize that these two receptors form a heteromeric complex in living cells. Here, we utilize three distinct biophysical and immunocytochemistry‐based approaches to identify and study this receptor complex in living cells. First, we generated a split luciferase complementation assay to test the hypothesis that the 5‐HT2AR and 5‐HT2CR are in close proximity (< 50 nm). We predicted that inactive luciferase fragments fused to each receptor would be brought into close proximity when receptors form a complex, thereby reconstituting the enzymatic activity of luciferase. A robust complementation (i.e., increased luminescence) was observed upon addition of the substrate D‐luciferin, an outcome that was dependent on protein levels assessed in radioligand binding studies (n=3, p<0.001). Co‐transfection with either the wildtype 5‐HT2AR or 5‐HT2CR inhibited the complementation (n=3, p<0.01). Using a proximity ligation assay, we demonstrate expression of the existence of 5‐HT2AR:5‐HT2CR heteroreceptor complexes in vitro, indicating the protomers are within 40 nm of each other (n=5, p<0.05). To further delineate the receptors are within very close proximity (< 10 nm) in living cells, we generated 5‐HT2AR and 5‐HT2CR constructs for use in a bioluminescence resonance energy transfer (BRET) assay. Preliminary results indicate that the 5‐HT2AR and 5‐HT2CR form a robust BRET interaction creating a maximal signal of 230 mBRET units and a BRET50 of 2.1, which was more than double the signal generated in control studies with the β2‐adrenergic receptor. Taken together, we have identified a very close 5‐HT2AR:5‐HT2CR interaction in living cells. Ongoing research will employ these technologies to further investigate the formation and function of the receptor complex.Support or Funding InformationNIDA T32 DA07287 “Neural and Pharmacological Mechanisms of Abused Drugs”This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Blood‐brain Barrier Penetrant and Orally Bioavailable Antidotes to Organophosphate Poisoning
Authors: Shyong Yan‐Jye , Garcia Arnold , Sepulveda Yadira , Radić Zoran , Momper Jeremiah , Taylor Palmer 
Published: June 2021

We have developed a series of antidotes to organophosphate poisoning, hydroxyiminoacetamido alkylamines, that ionize as cations, anions, zwitterions and form an appreciable neutral species at physiologic pH values. As such they cross the blood‐brain barrier and are orally bioavailable. They appear to be effective in mice and macaques in reversing organophosphate toxicity from accumulating sarin and paraoxon (Rosenberg et al., 2017, Chem‐Biol Interact 274, 50–57; Toxicol Lett. https//doi.org/10.1016/jtoxlet.2017.10.025). We examine here the blood and tissue disposition of these agents in the absence of organophosphate to ascertain pharmacokinetic profiles and achieve optimal loading and maintenance dosing of this family of antidotes. Our data show comparable absorption and tissue distribution after tail vein and retro‐orbital injection with only a slight delay when the antidote is administered intramuscularly. Hence, systemic distribution can be rapidly achieved to reverse acute toxicity. Initial studies of tissue disposition show both rapid transfer into the brain, along with rapid clearance from tissues and blood through likely transport mechanisms. Such kinetics are consistent with post‐administration Protection Ratios of the organophosphate after acute exposure and in vitro profiles of reactivation of the organophosphate inhibited enzyme. As with the quaternary oximes such as 2‐PAM and HI‐6, clearance is rapid requiring a parenteral loading dose and then parenteral or oral maintenance dosing to reverse or avert toxicity and fatality. Such studies in mice enable one to consider effective antidote treatment schemes for longer acting pesticides and acute exposure to the volatile sarin and related nerve agents through respiratory and dermal absorption routes.Support or Funding InformationSupported in part by NINDS, UO1‐058046This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Biasing μ Opioid Receptors with G Protein Inhibitors to Improve Opioid Analgesics
Authors: Campbell Adrian Phillip , Smrcka Alan V. 
Published: June 2021

The development of “safer” opioids is difficult owing to the fact that the undesirable effects of opioid analgesics are all on‐target effects at the μ opioid receptor (MOR). A new exciting approach to avoiding side effects has been development of “biased” MOR agonists that alter receptor conformation to preferentially activate analgesic pathways downstream, and away from pathways that promote undesirable side effects [1]. One such drug, Oliceridine, developed by Trevena pharmaceuticals has shown analgesic efficacy in acute pain with limited side effects in Phase IIb clinical trials. Our laboratory has pioneered an alternate approach to biasing GPCR signalling with small molecules that bind to the Gβγ subunits downstream of receptor activation to selectively modify signals downstream of GPCRs. Two prototype molecules, gallein and M119 potentiate the analgesic potency of morphine, without potentiating side effects typically associated with opioid use including acute tolerance, respiratory depression, constipation, hyperlocomotion, reward preference or physical dependence [2–4].To identify novel small molecule inhibitors of Gβγ with more drug‐like potential we screened a 140, 000 small molecule library comprised of diversity sets from Maybridge and Chemdiv. 36 compounds passed primary testing, triplicate retesting, dose response analysis, counterscreening for assay interfering compounds and triaging to exclude predicted aggregators, PAINS, toxic or reactive compounds and structures with limited drug‐like potential. The molecules have potencies of 2–100 μM in our primary Alphascreen assay (compared to 4 μM for the control SIGK peptide) and multiple compounds inhibit Gβγ activation of PLCβ in in vitro assays of PLC activity with potency equal to or better than the small molecule gallein. One compound has also shown potentiation of opioid analgesia similar to morphine in mice. The hits possess a variety of novel scaffolds and represent good starting points for further medicinal chemistry optimization and represent possible novel potentiators of opioid analgesia.Support or Funding Information5 R01GM0817720‐08This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Novel Approach to Safer Analgesics: Mu Opioid Receptor Agonist & Delta Opioid Receptor Antagonist Peptidomimetics
Authors: Fernández Thomas Joaquín , Griggs Nicholas , Nastase Anthony , Anand Jessica , Jutkiewicz Emily J. , Mosberg Henry , Traynor John 
Published: June 2021

Opioid analgesics exert their therapeutic and unwanted effects by activation of the mu‐opioid receptor (MOR). Repeated administration of opioid analgesics and subsequent chronic MOR activation promotes opioid tolerance and physical dependence. It has been proposed that activation of MOR with concurrent antagonism of the delta‐opioid receptor (DOR) can attenuate the development of opioid tolerance and dependence. To further investigate the utilization of concurrent MOR agonism and DOR antagonism we designed, synthesized, and characterized a library of “peptidomimetics” that have elements of opioid peptides vital for activity but retain small molecule‐like features necessary for blood‐brain‐barrier penetration. Compounds were evaluated in membrane preparations of cloned cell lines expressing either MOR, DOR, or kappa‐opioid receptors (KOR). [35S]‐GTPγS binding to measure G protein‐coupled activation was used. Full agonists at each opioid receptor (DAMGO, DPDPE, and U69,593, respectively) were employed as standards. Maximum binding and potency (EC50) values were determined from concentration‐effect curves. To measure affinity (Ki) values, competition binding using [3H]‐diprenorphine was performed. Peptidomimetics that demonstrated promising bi‐functional activity were tested for antinociceptive activity in vivo. Compared to repeated morphine‐treated mice, repeated peptidomimetic treated mice developed less analgesic tolerance or physical dependence. Analysis of in vitro and in vivo data has permitted structure‐activity relationships to guide further chemical synthesis and the discovery of opioid analgesics that may have improved clinical utility.Support or Funding InformationSupported by DA‐03910This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.