bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neonatal Inflammation Impairs Multiple Pathways to Adult Respiratory Plasticity
Authors: Hocker Austin Dean , Huxtable Adrianne G. 
Published: June 2021

Bacterial‐induced inflammation is a common clinical problem in neonates and acutely impairs respiratory control, but we know little about the long‐term effects of neonatal inflammation on adult respiratory control. Since neonatal inflammation blunts adult immune function, reduces adult hippocampal plasticity, and increases the vulnerability to neuropsychiatric disorders, we hypothesized that neonatal systemic inflammation would inhibit adult phrenic long‐term facilitation (pLTF), a form of respiratory neuroplasticity induced by acute intermittent hypoxia (AIH). pLTF can be induced by distinct cellular mechanisms including the Q‐pathway, induced by moderate AIH (PaO2 35–45 mmHg), and the S‐pathway, induced by severe AIH (PaO2 25–35 mmHg). Therefore, we tested the hypothesis that neonatal inflammation would undermine both the Q‐ and S‐pathway to pLTF. Postnatal day 4 (P4) pups of both sexes were injected with either lipopolysaccharide (LPS; 1 mg/kg) or sterile saline and allowed to mature to adulthood. In order to control the known effects of estrus cycle hormones on pLTF, adult females were ovariectomized 7–8 days prior and given 17‐β estradiol (40 μg/ml/kg, 3 hours) before AIH. Neonatal LPS undermined adult mAIH‐induced (Q‐pathway) pLTF in both females (−15 ± 17%, n=6, p < 0.0001) and males (14 ± 14%, n=12, p = 0.01) relative to neonatal Saline controls (females; 97 ± 24%, n=7, males; 55 ± 13%, n=7). Acute, adult treatment with a systemic anti‐inflammatory (ketoprofen, 12.5 mg/ml/kg, i.p., 3 hours) restored Q‐pathway pLTF after neonatal LPS in both females (118 ± 24%, n=4) and males (58 ± 9%, n=4) to saline controls treated with ketoprofen (P4 Saline females; 67 ± 11%, n=4, males; 54 ± 9%, n=4). Additionally, sAIH‐induced (S‐pathway) pLTF was undermined in adults after neonatal LPS in females (0 ± 17%, n=4, p < 0.001) and males (7 ± 9%, n=4, p < 0.01) compared to neonatal saline control females (86 ± 23%, n=4) and males (47 ± 17%, n=4). However, systemic ketoprofen in adults did not restore S‐pathway pLTF after neonatal inflammation in females (P4 Saline; 136 ± 11%, n=4, P4 LPS; 23 ± 15%, n=5, p < 0.0001) or males (P4 Saline; 67 ± 11%, n=4, P4 LPS; −5 ± 37%, n=4, p < 0.01). Thus, neonatal systemic inflammation has a lasting effect of respiratory plasticity by abolishing multiple pathways to adult respiratory plasticity. Furthermore, the restoration of Q‐pathway, but not S‐pathway, mediated plasticity by acute ketoprofen demonstrates different mechanisms impair adult respiratory plasticity after neonatal inflammation. Understanding how neonatal inflammation impairs the adult respiratory system will elucidate links between neonatal conditions and adult respiratory insufficiency, leading to better therapies to enhance breathing in all age groups.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activation of Airway Afferent Vagal C‐fibers by Sphingosine‐1‐Phosphate
Authors: Patil Mayur Jayanta , Meeker Sonya , Undem Bradley 
Published: June 2021

Airway inflammation leads to a release of myriad inflammatory mediators and autacoids. At the site of inflammation these molecules in theory can activate airway C‐fibers leading to the observed symptoms of reflex bronchoconstriction, mucus secretion, as well as sensations of dyspnea and urge‐to‐cough. Relatively little progress has been made to understand which among the plethora of mediators that arise during airway inflammation are most responsible for C‐fiber activation. Here we show that shingosine‐1‐phosphate, a lipid that is elevated during airway inflammatory conditions like asthma, strongly activates airway vagal C‐fibers. We employed an ex‐vivo mouse trachea/lung‐vagus nerve preparation in which action potential discharge in single afferent nerves terminating in the lung can be evaluated with extracellular recording from an electrode placed in the vagal sensory (nodose/jugular) ganglion. Drugs were delivered into the lungs directly to the receptive field. S1P (10μM) strongly stimulated, in a concentration dependent manner, 62% (n=21 out of 34 tested) of vagal C‐fibers terminating in the trachea/lungs (CV=0.63±0.04m/s), (426±71 APs, 7.2±4 Hz peak Freq). An antagonist to S1Pr3, TY 52156 (10μM) significantly blocked the stimulatory effect of S1P on S1P positive C‐fibers (214±53 APs in control vs 56±18 APs in presence of TY 52156, n=7 p=0.01). We developed an imaging technique using 2‐photon microscopy and Pirt‐GCamp6 mice to simultaneously monitor the activity of hundreds of vagal ganglion neurons in the trachea/lung‐ganglia preparation upon drug delivery into the lungs. About 40% of capsaicin (1μM) positive neurons responded to S1P (10μM). Single cell RT‐PCR revealed that ~90% of nodose ganglia neurons retrogradely labeled from the airways that express TRPV1 (denoting C‐fiber neurons) also express the S1P receptor, S1Pr3. In an RNA seq transcription analysis specifically of capsaicin‐sensitive nodose neurons we found S1Pr3 was selectively and strongly expressed. In counts per million the mRNA expression was 0, 8, 376, 3, 0 for S1Pr1, 2, 3, 4 and 5, receptors respectively. Jugular neurons expressed relatively little S1Pr receptors. We hypothesize that S1P may contribute to vagal (nodose) C‐fiber activation in airway inflammatory diseases.Support or Funding InformationRO1 HL137807This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Probing RNA Structure and Dynamics in the HIV‐1 5′ UTR using Ensemble and Single Molecule Approaches
Authors: Musier‐Forsyth Karin , Brigham Benjamin , Kitzrow Jonathan , Reyes Joshua‐Paolo C. , Munro James 
Published: June 2021

One of the most conserved and highly structured regions of the HIV‐1 genome, the 356‐nt 5′UTR, acts as a scaffold for many protein and nucleic acid binding partners and regulates vital processes including initiation of reverse transcription, genomic RNA (gRNA) dimerization and packaging. Previous studies support the presence of at least two mutually‐exclusive 5′UTR conformational states (Huthoff and Berkhout, RNA, 7, 2001; Lu et al., Science 334 (2011), but whether the 5′UTR undergoes a conformational switch during the viral lifecycle and what regulates this switch are less clear. In‐gel selective 2′‐hydroxyl acylation followed by primer extension (SHAPE) probing studies of the wild‐type (WT) 5′UTR revealed a mixed population with structural differences between monomer and dimer, consistent with a structural switch (Kenyon, JC et al, Nucleic Acids Res 41, 2013). However, these studies did not address the dynamics of the conformational change or probe possible triggers. In this work, we used single‐molecule Förster resonance energy transfer (SM‐FRET) studies to probe 5′UTR (ΔTAR/polyA) conformational dynamics and the factors that modulate it. Intra‐molecular FRET assays showed that mutations in the dimerization initiation site (DIS) and U5 region of the monomeric RNA modulated the conformational equilibrium in the expected manner, with the ΔDIS construct shifted to a high‐FRET dimerization‐competent state. Increasing Mg2+ concentration promoted the dimer‐competent conformation in the WT 5′UTR. In 10 mM Mg2+, the WT construct occupies both the high‐FRET (39%) and low‐FRET states (61%). Individual FRET traces showed that of the 841 molecules examined under these conditions, 29% transitioned between FRET states during the ~150 sec observation window. Inter‐molecular FRET observed between an immobilized donor‐labeled 5′UTR construct and excess acceptor‐labeled 5′UTR was consistent with formation of a stable extended dimer. In‐gel FRET assays were also consistent with the formation of a stable dimer. Surprisingly, tRNA‐annealing to the primer binding site of an immobilized 5′UTR promoted the monomer conformation even in the presence of high Mg2+, as observed by both SM‐FRET and native PAGE. Overall, these data are consistent with high intrinsic conformational dynamics in the 5′UTR that can be regulated by factor binding.Support or Funding InformationNational Institutes of HealthThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Selenoprotein profiling in various tissues of mice fed with selenium‐deficient and high‐selenomethionine diets.
Authors: Ishii Isao , Akahoshi Noriyuki , Kamata Shotaro , Hashimoto Yuri , Hayashi Seiya , Tokoro Natsumi , Yamamoto Shingo , Shimada Ki‐ichi , Anan Yasumi 
Published: June 2021

Selenium (Se) is a trace essential nonmetal element and its deficiency is known to cause myocardial necrosis leading to the heart weakening (e.g. Keshan disease). Se deficiency may contribute to Kashin‐Beck disease, resulting in atrophy, degeneration, and necrosis of cartilage tissue in the joint, or hypothyroidism including extreme fatigue, goiter, mental retardation, and miscarriages. In contrast, Se excess may cause reversible balding and brittle nails, give a garlic odor to the breath, and cause intestinal distress, weakness and slowed mental functioning. All such clinical symptoms could be attributable to dysregulated expression of twenty‐five selenoproteins in humans; mice also have the corresponding 25 selenoproteins. This study examined expression of the representative 11 selenoproteins (Gpx 1/2/3/4, TrxR1/2, Dio1, Sephs2, SelP, SelS, and SelN) in 8 different organs (brain, heart, liver, lung, kidney, pancreas, spleen, and testis) of mice fed ad libitum selenium‐deficient and high‐selenomethionine (a major dietary source from plants) diets. One week with the diets caused significant decreases and increases in Se contents of each organ, respectively, which became slightly more apparent until 4 weeks. Se deficiency in diet induced significant decreases in Gpx1/2 of liver, kidney, and pancreas; however, neither Se deficiency nor selenomethionine excess caused significant expression changes in other selenoproteins of all organs tested. There results indicate that altered organ Se contents upon Se deficiency or excess in short periods (~4 weeks) do not generally reflect selenoprotein expression levels in these organs.Support or Funding InformationSupported by the Grants‐in‐Aid for Scientific Research (16H05107, 16K15194, 17K08287, and 25220103) from the MEXT of Japan.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protein E6 in High Risk Human Papillomaviruses
Authors: Mahesh Rhea , Langat Daisy , Langat Diana , Barbara Ella , Cheng Amanda 
Published: June 2021

Human Papillomavirus (HPV) refers to a group of more than one hundred HPV high risk or low risk strains associated with a variety of conditions. Over fifty strains lead to genital infections, while others influence the development of various cancers, such as cervical, skin, etc. The combination of HPV E6/E7, hTERT, and Ki67 mRNA expression levels complement each other to diagnose lesions of the aforementioned cancers, specifically dealing with the cervix. All HPV strains share a common structure: circular, double stranded DNA encoding for eight open reading frames (ORF) surrounded by a non‐enveloped capsid. ORFs function to produce the E1‐E7 proteins needed for HPV replication (early region), L1‐L2 proteins that make up the capsid (late region), and replicate and transcribe viral DNA with the help of cis components (long control region). The E6, in particular, is an oncoprotein composed of polypeptides made up of 150 amino acids and a Cys‐X‐X‐Cys motif that allow for the formation of two zinc fingers. E6 and E7 recognize a number of host proteins by hijacking their cellular domain‐motif interaction networks. In the HPV‐16 and HPV‐18 strains, the E6 protein binds its amino and carboxyl terminal regions to a leucine‐rich sequence (LxxLL) within the cellular ubiquitin ligase, E6AP. This heterodimer degrades p53, a proapoptotic tumor suppressor, although the exact mode of assembly is unknown. Complete degradation potentially leads to oncogenesis in high‐risk HPV strains, as these transformed cells have lost their ability to arrest at the G1 checkpoint within their cell cycle and perform apoptosis. Aside from the E6‐E6AP heterodimer binding directly to p53, the E6 protein has other oncogenic functions, allowing it to bind to four general classes of other proteins, providing indirect pathways to degrade p53. Binding options include proteins involved in cell polarity and motility, tumor suppressors and inducers of apoptosis, transcriptional coactivators and DNA replication/repair factors. Regarding transcription coactivators, degradation of p53 transcriptional activity is possible, as seen by the targeting of p53 coactivator p300/CBP or yADA3. Given the mechanism of E6‐E6AP complex hindering p53's from carrying out normal function, E6 is an attractive target for therapies against cervical cancers. The Olathe North High School MSOE Center for BioMolecular Modeling SMART Team used 3‐D modeling and printing technology to examine structure‐function relationships of E6.Support or Funding InformationSMART team is a joint venture between the Milwaukee School of Engineering and the Olathe School District's 21st century program Medical Professions Academy.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluating the nuclease activity of Pseudomonas exotoxin A
Authors: Patel Nirja , Jackson Namo , Weldon John 
Published: June 2021

Pseudomonas exotoxin A (PE) is a protein toxin secreted by Pseudomonas aeruginosa. The toxin is remarkably potent, and believed to be the major virulence factor of P. aeruginosa. PE inhibits protein synthesis by inactivating elongation factor 2 (EF2), the protein that advances the ribosome along the mRNA during translation. Cell with inactive EF2 are unable to synthesize proteins, which leads to apoptosis. Recent studies have indicated that, in addition to inhibition of protein synthesis, PE has also functions as a nuclease. The purpose of the experiments described here is to verify this unusual claim and characterize the nuclease activity of PE using only the catalytic fragment of the toxin. We have conducted basic time‐course and concentration assays, and have confirmed a low level of nuclease activity from PE. We are currently examining cleavage in the presence of different divalent cations (Ba2+, Sr2+, Mn2+, Co2+, Ca2+, Mg2+ and Zn2+). We have also tested a fluorescence‐based assay to observe cleavage of DNA in real‐time. Initial experiments using DNase I have been successful, and we hope to use this method to study PE in the near future.Support or Funding InformationResearch reported in this poster was supported by the National Institute of General Medical Sciences of the National Institutes of Health under award number R25GM058264. Additional support for this research comes from Towson University, the Jess and Mildred Fisher College of Science and Mathematics, Graduate Studies and the Department of Biological Sciences.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exploration of membrane binding of α‐synuclein through Site Based Spin Labeling
Authors: Vrabel Jarret 
Published: June 2021

Neurodegenerative conditions, including Alzheimer's and Parkinson's disease, are becoming more prevalent in the aging population, and one of the main perpetrators behind these symptoms is the protein α‐synuclein (α‐syn). The cytosolic neural protein has a role in neurotransmitter transport within cells through membrane binding to intracellular vesicles. It is strongly linked to Parkinson's disease as a result of the A30P, E46K, and A53T point mutations, which are believed to cause α‐syn aggregation. These point mutations disrupt the formation of amphipathic α‐helices that directly bind to the vesicle surface. To gain further information on the membrane binding tendencies of α‐syn, each of these mutations, as well as a neutrally mutated C‐terminal tail (CTT), is being separately tested using site directed spin labeling and electron paramagnetic resonance (EPR) spectroscopy. The normally negatively charged CTT is the only region of the protein that does not directly contact the phospholipids during the membrane binding. Each of the mutant proteins' interactions with synthesized lipid vesicles will confirm the known malfunctions of the Parkinson's mutations, in addition to expanding of knowledge of the C‐terminal tail and its function in the membrane binding process. Results and analysis from the EPR spectroscopy are currently still being compiled. It is expected that each of the Parkinson's mutations and neutrally mutated CTT will decrease α‐syn membrane binding. These, or other conclusions from the analysis, can provide more information on the uncommonly studied α‐synuclein CTT and a basis on which to continue research for future undergraduate biochemistry students at Westminster College.Support or Funding InformationWestminster College Drinko Center Undergraduate Research GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
IMPACT OF BETA‐DYSTROGLYCAN DEFICIENCY ON EMERIN FUNCTIONS
Authors: Méndez Jesús Mauricio Ernesto Hernández , Monsiváis Wendy Lilián Gómez , González Ricardo Mondragón , Perlingeiro Rita , Mena Jaime García , Vega Bulmaro Cisneros 
Published: June 2021

Beta‐dystroglycan (b‐DG) is a plasma membrane protein that belongs to the dystrophin‐associated protein complex (DAPC), the latter provides a link between the extracellular matrix and the cytoskeleton. Previously we reported that b‐DG is translocated localized to the nucleus via recognition of its nuclear localization signal (NLS), located in the yuxtamembrane domain, by importins α2/β1. Nuclear b‐DG is involved in the maintenance of nuclear envelope structure through its interaction with emerin and lamins A/C and B1. Emerin is an inner nuclear membrane protein involved in nuclear morphology, myogenesis and cell proliferation.In this study, we analyzed the impact of β‐dystroglycan deficiency on emerin function, using C2C12 cells as model. We found that depletion of β‐DG resulted in decreased emerin levels, as shown by immunofluorescence and western blot assays with anti‐emerin antibodies. Furthermore, emerin‐mediated processes, including nuclear morphology and nucleus‐centrosome distance are altered in such cells. The analysis of emerin‐dependent nuclear signaling in β‐DG‐depleted cells is currently in progress.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Arkadia (RING finger protein 111) mediates sumoylation‐dependent stabilization of Nrf2 through K48‐linked ubiquitylation
Authors: Davis Jamaine , McIntosh Deneshia , Walters Treniqka , Arinze Ifeanyi 
Published: June 2021

The transcription factor Nrf2 is a master regulator of the antioxidant defense system that serves to protect cells from oxidative damage. We previously reported that the SUMO‐targeted E3 ubiquitin ligase (STUbL), RING finger protein 4 (RNF4) accelerated the degradation rate of Nrf2 in promyelocytic leukemia‐nuclear body (PML‐NB)‐enriched fractions and decreased Nrf2‐mediated gene transcription. STUbLs contain SUMO‐interaction motifs that enable it to target substrate proteins that have been modified by SUMO resulting in ubiquitylation. These ubiquitylation events help in regulating eukaryotic cellular activity. In this study, we establish the second mammalian STUbL, Arkadia/RNF111 ubiquitylates polysumolyated Nrf2 to stabilize it, but has no effect on Nrf2‐mediated gene transcription. Interestingly, we discovered that the Arkadia‐mediated stabilization of Nrf2 occurs through Ub‐K48 linkages rather than the predicted Ub‐K63 linkages. These results suggest Arkadia‐mediated ubiquitylation of Nrf2 protects it from degradation, thereby allowing Nrf2‐dependent gene transcription. Collectively, these findings highlight a novel mechanism to positively regulate nuclear Nrf2 levels in response to oxidative stress.Support or Funding InformationThis work was supported by NIH grant SC1CA143985 and by CTSA award No. ULITR000445 from the National Center for Advancing Translational Sciences. +Supported by NIH grant # 2 S21MD000104This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phosphorylation of SLIMB by Minibrain/DYRK1a Activates SLIMB‐mediated Circadian Clock Protein Degradation
Authors: Contreras Adam , Tabuloc Chrisitne , Li Ying , Vanselow Jens , Schlosser Andreas , Chiu Joanna 
Published: June 2021

SLIMB/β‐TrCP is a F‐box protein that provides substrate specificity to E3 ubiquitin ligase protein complexes, which ubiquitinate cellular proteins and target them for degradation by the 26S proteasome. Timely degradation of substrate proteins is intricately linked and critical to the regulation of numerous molecular processes, including cell cycle regulation, developmental timing, tumorigenesis, metabolism, and circadian biology. Previous work in our laboratory characterized the requirements for the interaction between SLIMB and PERIOD (PER), a key transcription factor that regulates circadian gene expression. The level of PER oscillates in a circadian manner and loss of SLIMB function disrupts PER cycling, leading to misregulation of the circadian oscillator. In contrast to the numerous studies examining the post‐translation modification (PTM) of substrate proteins to enable recognition by SLIMB, e.g. interaction of SLIMB and PER, there is a deficit of literature that investigates the PTM of F‐box proteins that modulate their function. To understand the post‐translational regulation of SLIMB function, we have identified 7 phosphorylation sites on SLIMB using mass spectrometry proteomics and characterized the function of these SLIMB PTMs in the context of the Drosophila circadian clock system.Support or Funding InformationNIH R01 GM102225 to JCC, F31 DK104624 to AJC, and T32 GM007377This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.