bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cotinine enhances the nicotine‐mediated decrease in respiratory burst amplitude recorded from the hypoglossal nerve in the brainstem spinal cord preparation from nicotine‐exposed neonatal rats
Authors: Flanigan Emily Gayle , Fregosi Ralph Frank , Wollman Lila Buls 
Published: April 2020

Cotinine, the primary metabolite of nicotine, can be detected in the blood of infants born to mothers who used nicotine products during pregnancy and breast feeding, and is used as a marker of nicotine exposure. Previous research shows that prenatal nicotine exposure alters the regions of the brain that control breathing and results in impaired protective breathing reflexes in exposed human and animal neonates. These abnormalities are proposed as a mechanism to explain Sudden Infant Death Syndrome (SIDS), and maternal smoking is the number one risk factor for SIDS. While it has long been known that nicotine has direct effects on brain function and development, it has only recently been established that cotinine may act on the brain as well. Both nicotine and cotinine bind to nicotinic acetylcholine receptors (nAChRs) which are expressed on neurons throughout the nervous system. Preliminary data using the in vitro brainstem spinal‐cord preparation from nicotine‐exposed neonatal rats shows that activation of nAChRs produces a decrease in the amplitude of respiratory bursts recorded from hypoglossal nerve roots (−30.74±18.2 % baseline, n=4), and that this effect is enhanced by cotinine (−60.34±16.1 % baseline, n=4). This supports the hypothesis that cotinine modulates the function of nAChRs in regions of the brain that control breathing (P<0.0001).Support or Funding Information5R01HD071302‐07
Effects of the G Protein‐coupled Estrogen Receptor on Cardiovascular Remodeling Induced by Chronic Sympathetic Activation
Authors: Tan Derick , Twohey Stephanie , Allen Patrick , Giles Jennifer , Winter Lori , Clayton Sarah , Tran Quang-Kim , Eric Wauson 
Published: April 2020

Estrogens modulate cardiovascular function via the classical nuclear hormone receptors ERα and ERβ, and the more recently discovered G protein‐coupled estrogen receptor (GPER). While the activation of GPER elicits many cardioprotective effects, the mechanisms behind these effects are incompletely understood. Sympathetic nervous activities are essential for normal cardiac functions; however, chronic sympathetic activation is an important contributor to maladaptive cardiovascular remodeling that leads to cardiovascular diseases. Our recent data demonstrate that GPER activity moderates the increases in myocardial contraction and calcium dynamics stimulated by acute activation of the cardiac beta1 adrenergic receptor. However, the effects of GPER activation on the cardiac manifestations in response to chronic sympathetic activation are unknown. We hypothesized that GPER activation would reduce cardiovascular remodeling induced by chronic norepinephrine (NE) administration. In adult female ovariectomized mice, administration of NE (10 mg/kg/day) via osmotic pumps for 8 weeks increased heart weight, kidney weight, cardiomyocyte size as assessed by wheat germ agglutinin staining, left ventricular wall thickness, and aorta wall thickness. These changes occurred without an apparent increase in blood pressure assessed by tail cuff measurements. These effects were reduced by the co‐administration of GPER agonist G‐1 (120 mg/kg/day), while administration of G‐1 alone at the same dose had no effect. These initial data indicate that the GPER agonist G‐1 can reduce cardiovascular remodeling caused by chronic sympathetic activation.Support or Funding InformationDMU IOER 05‐19‐06
Evaluation of FDA Drugs to Identify Modulators of Multidrug Resistance Associated Protein 1 (MRP1) Expression Levels in HEK293 MRP1 Overexpressing Cells.
Authors: Loecker Josh , Poku Vivian Osei , Iram Surtaj 
Published: April 2020

The Multidrug Resistance Associated Protein 1 is a key feature in drug resistance within cancerous cells. This protein is involved in efflux of a broad spectrum of substrates, including organic anions and several chemical therapeutic agents, like doxorubicin. In addition, MRP1 has implications in separate physiologies such as cardiovascular disease and age‐related macular degradation. As such, its overexpression has been implicated in multidrug resistance of several cancers.This project is aimed at evaluating the effect of chemotherapeutic agents on the expression levels of MRP1 in HEK293/MRP1 cells. A unique library of FDA drugs were screened using a high throughput In‐Cell ELISA Assay. A total of 88 drugs were screened. Drugs that showed modulation of MRP1 protein expression above 30% were considered as Hit compounds. A significant number of hits were identified, from which some of the drugs increased MRP1 protein expression and decreased its expression.This protein enhances the cells ability to efflux toxic compounds, such as anticancer drugs, out of the cell. A cell’s ability to increase expression of this protein results from a genetic mutation. As a result, higher concentrations of traditional anticancer drug compounds are required to demonstrate similar results as cells that do not over express MRP1. However, discovery of drugs that can decrease the expression of this protein would play a valuable role in assisting the fight against cancer.Support or Funding InformationI would like to thank Jack and Mary East for their support and funding for this project
Cyclooxygenase Inhibition Increases the Sympathetic Response to Hypercapnia
Authors: Eisenmann Nicole A. , Howery Anna J. , Miller Kathleen B. , Eldridge Marlowe W. , Barnes Jill N. 
Published: April 2020

Cyclooxygenase (COX) catalyzes the formation of prostaglandins and influences blood flow regulation. Previous studies have shown that COX inhibition reduces cerebral blood flow velocity and cerebrovascular reactivity to CO2 (CVR), however, the effects of COX inhibition on neurovascular regulation is unknown. Sympathetic nerve activity is an important modulator of blood pressure and blood flow. The purpose of this study was to determine the effect of COX inhibition on the muscle sympathetic nerve activity (MSNA) response to hypercapnia during CVR testing.MethodsFifteen healthy adults were evaluated (M/F = 7/8; age = 40 ± 21 years; BMI = 24 ± 2 kg/m2) before (Pre) and 90 minutes after oral administration of the COX inhibitor indomethacin (Indo), given at 1.2mg/kg. Mean arterial pressure (MAP), heart rate (HR) and MSNA were measured continuously throughout the protocol. After baseline measurements were recorded, participants were evaluated breathing 6% CO2. Results are reported as mean ± SE.ResultsAt baseline, HR was 54 ± 2 bpm, MAP was 100 ± 4 mmHg, MSNA burst frequency was 24 ± 2 bursts/min and MSNA burst incidence was 45 ± 4 bursts/100hb. The change in HR when exposed to hypercapnia was not different after Indo (Pre: 6 ± 2 bpm vs. Indo: 4 ± 1 bpm; p>0.05). The change in MAP (Pre: 0 ± 2 mmHg vs. Indo: 6 ± 1 mmHg; p<0.05), MSNA burst frequency (Pre: 1 ± 1 bursts/min vs. Indo: 4 ± 1 bursts/min; p<0.05), and MSNA burst incidence (Pre: −3 ± 2 bursts/100hb vs. Indo: 4 ± 2 bursts/100hb; p<0.05) was greater after Indo administration.ConclusionThe sympathetic response to hypercapnia was greater following the administration of Indo. Taken together, these results suggest that hypercapnia‐induced sympathoexcitation may be altered following COX inhibition. Future studies could evaluate if the sympathetic responses to hypercapnia vary with age or sex.Support or Funding InformationNIH HL118154
Precisely Targeted Reporters Reveal ABCG2 Regulation in Live Lung Cancer Cells
Authors: Kovacsics Daniella , Brózik Anna , Tihanyi Borbála , Matula Zsolt , Borsy Adrienn , Mészáros Nikolett , Szabó Edit , Németh Eszter , Fóthi Ábel , Zámbó Boglárka , Szüts Dávid , Várady György , Orbán Tamás I. , Apáti Ágota , Sarkadi Balázs 
Published: April 2020

Expression of the ABCG2 multidrug transporter is a marker of cancer stem cells and an indicator of poor disease outcome. Understanding the effect of pharmacotherapy on ABCG2 expression is critical in guiding therapy recommendations and may help rational drug development. Engineered reporter cell lines are useful in monitoring gene regulation and protein activity in live cells but rely on precise targeting to preserve native regulatory functions. Here, we generated a number of precision‐engineered, fluorescent human cell lines that interrogate ABCG2 regulation in live lung cancer cells. Using homology‐based CRISPR engineering, we targeted a promoterless eGFP to the translational start site of the ABCG2 gene, generating ABCG2 knockout and in situ tagged ABCG2 reporter cells. Using the engineered cell lines, we show that anti‐cancer medications, HDAC inhibitors and hypoxia‐mimicking agents induce ABCG2 expression. In addition, we provide evidence that the glucocorticoid receptor is a specific, positive regulator of the ABCG2 gene. To our knowledge, this is the first fluorescent reporter assay designed to follow the endogenous regulation of a human ABC transporter in live cells. The information gained may direct therapy recommendations and assist rational drug design.Support or Funding InformationThis research has been supported mainly by OTKA‐NKFIH grant NK 115375, but also by NKFIH K104903 and the NVKP_16‐1‐2016‐0005 program from the National Research, Development and Innovation Office. This work has also been supported by research grant OTKA‐NKFIH K128011, awarded to GV. A Borsy was supported by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences (HA: BO/00579/17/5), through the New National Excellence Program of the Ministry of Human Resources (HA: ÚNKP‐18‐4‐SE‐11).
The Development of an Educational e‐Module to Teach the Pathogenesis of Cancer Utilizing an Anatomical Donor Case Study
Authors: Fortney Lane , Kowalewska Jolanta , Kolheffer Kerwin , Elzie Carrie , Wellman Laurie 
Published: April 2020

IntroductionThere is a significant need for understanding the complexity and gravity of cancer pathogenesis as it continues to be a leading cause of death worldwide. During a gross anatomy lab, students encountered a rare malignancy, Uterine Leiomyosarcoma, in one of the donors. After further investigation and confirmation of the diagnosis, this experience was used to develop an educational e‐module to teach cancer pathogenesis.MethodologyRelevant measurements and photographs were obtained, as well as representative sections of the uterine tumor specimen and distant metastases. The Eastern Virginia Medical School Biorepository performed histological preparations from these tissues, including standard hematoxylin and eosin preparations, as well as staining for smooth muscle actin (SMA). A faculty pathologist confirmed the diagnosis of leiomyosarcoma, applying current inclusion criteria. The Storyline Articulate 360 software platform was used to generate a case‐based interactive e‐module with pre‐ and post‐test questions addressing the concepts of cancer pathogenesis, tumor heterogeneity, cancer nomenclature, lineage of differentiation, and cancer classification, relating these topics to the findings and likely clinical course of this case.ResultsThe e‐module was beta tested for usability and accuracy with a focus group of faculty members who provided qualitative and quantitative feedback. The resulting e‐module will be implemented into a general pathology course taken by medical and graduate students in the spring semester of 2020. Data on usability, knowledge, and application of an anatomical donor case for understanding cancer pathogenesis will be collected. The outcomes of this process will be evaluated for future curriculum implementation.ConclusionThe interactive e‐module developed is an effective pedagogical tool to reinforce students’ knowledge of cancer pathogenesis using an anatomical donor case study. Inspired by a pathological finding in a gross anatomy lab, this interdisciplinary educational opportunity integrates and enhances a pathology course from a dissection experience. The integration of this interactive e‐module into medical school curriculum has the potential to be a powerful resource for understanding the fundamental principles of neoplasia through a novel approach of utilizing findings from an anatomical donor.Support or Funding InformationSupport and Funding provided by the Contemporary Human Anatomy Program at Eastern Virginia Medical School
Pilot Evaluation of Telemetered Cardiac Hemodynamics in the Setting of Fully‐Implanted Programmable Drug Delivery in Rats: A New Tool for Cardiovascular Drug Discovery?
Authors: Bennett Nikki Luis , Zambataro Christopher L. , Velie Troy , Bogie Heather , Shahidi-Latham Sheerin Luis , Ferguson Brian , del Rio Carlos Luis 
Published: April 2020

IntroductionThe mechanistic understanding of pharmacokinetics–pharmacodynamics (PK/PD) relationships is a key component of modern cardiovascular drug discovery. However, given that both PK properties and/or experimental techniques (such as anesthesia) can alter auto‐regulatory/autonomic control in vivo (thus masking true PD responses), tools permitting the determination of translationally‐relevant PK/PD relationships for cardio‐active agents are limited. This pilot study established the feasibility of combining implantable programable infusion pumps and telemetered cardiac hemodynamic monitoring for the assessment PK‐independent PD responses in untethered conscious rats.MethodsHealthy Sprague‐Dawley rats (n = 7) were chronically instrumented with both a digital telemetered micro‐manometer catheter (HD‐S21, Data Sciences International) in the LV, and with a fully‐implanted programmable/refillable infusion pump in a femoral vein (iPRECIO® SMP‐200). The pumps were programmed, a priori, to perform (after a 21‐day recovery period) a two‐arm drug delivery protocol, consisting (each arm) of a 72h active period (ON, @12.5μL/hr) followed by a 96h washout (OFF period, on patency mode @1μL/hr); prior to each active period the pumps were loaded with either vehicle (CTRL) or a novel pure negative inotrope (MYK‐581), while normal saline was loaded prior to OFF periods. Left‐ventricular pressures (LVP) and derived functional indices were evaluated continuously (via telemetry) before/during active periods, while cardiac function/geometry were assessed (via echocardiography) prior to the end of the active period.ResultsFully implanted conscious rats had normal cardiac hemodynamics (HR: 339 ± 8 bpm, LVPmax: 119±3 mmHg) and cardiac function (EF: 76 ± 2%, FS: 47± 1%, and dLVP/dtmax: 9275 ± 369 mmHg/s), showing normal circadian oscillations. Similar observations were made during vehicle administration (e.g., EF: 77 ± 1%, and dLVP/dtmax: 9585 ± 891 mmHg/s), while marked reductions in both echocardiographic (EF: 45 ± 8.7%, FS: 24 ± 6%, P < 0.05 vs. VEH) and LVP‐derived (dLVP/dtmax: −21 ± 6%, or 7313 ± 819 mmHg/s, P < 0.05 vs. VEH) functional indices were observed during the MYK‐treatment period.ConclusionsTaken together, these data demonstrate that telemetered hemodynamics and fully‐implanted programmable/refillable infusion pump technologies can be successfully integrated in vivo in order to generate translationally‐relevant PK‐independent pharmacodynamics data in untethered conscious rats.Support or Funding InformationThese studies were supported by MyoKardia
Pyruvate Kinase M2 Deficiency Promotes Brown Fat Adipogenesis in Vitro and Enhances Adaptive Thermogenesis in Vivo
Authors: Alquraishi Mohammed , Puckett Dexter L. , Chahed Samah L. , BAETTAIEB AHMED 
Published: April 2020

The prevalence of obesity and its comorbidities has sparked a worldwide concern to address rates of adipose tissue accrual. Pharmacological approaches to combat obesity have met with limited success despite intensive investments in their development. Although many differing strategies are being explored in cellular and molecular studies, many have proven ineffective at achieving long‐term positive outcomes. Thus, the search for novel approaches and new target must continue. Brown adipocytes are specialized to dissipate chemical energy as heat and are associated with improved metabolic phenotypes. Herein we demonstrate that altered expression of the glycolytic enzyme pyruvate kinase M2 (PKM2) promotes adipogenesis of mouse and human brown adipocytes. Additionally, modulation of PKM2 expression results in increased lipolysis through modulation of the activity of protein kinase A and hormone‐sensitive lipase (HSL). Importantly, PKM2 deficiency in brown adipocytes increases ATP turnover, and enhances basal and maximal mitochondrial respiration. Furthermore, mice with specific deletion of PKM2 in brown adipose tissue exhibited increased energy expenditure and resistance to cold‐induced alteration in core body temperature. Collectively, these findings identify PKM2 as a novel component of the molecular circuit that contributes to brown fat adipogenesis and adaptive thermogenesis, which may have potential therapeutic implications.Support or Funding InformationThis work was supported by the National Institute of Diabetes and Digestive and Kidney Diseases (R00DK100736) to A.B.
Renal blood flow and oxygen metabolism in renovascular hypertension
Authors: Patinha Daniela , Koeners Maarten P. 
Published: April 2020

Renovascular hypertension affects ~1–4% of the hypertensive population and represents the second‐leading cause of secondary hypertension. There is an urgent clinical need to gain insight into the aetiology of renovascular hypertension to form the basis of new treatment targets. We hypothesize that high blood pressure in these conditions aims to sustain perfusion thus optimising organ blood flow and tissue oxygen homeostasis.Male Wistar Rats with (2K1C, n=5) or without (control, n=3) renal artery clip to generate renovascular hypertension were used. Six weeks after renal artery clipping, renal hemodynamics and oxygen metabolism were evaluated under isoflurane anaesthesia. Renal blood flow was measured in the clipped kidney using an ultrasound probe, glomerular filtration rate was measured using FITC‐inulin and the arterio‐venous difference in oxygen content was used to determine the clipped kidney oxygen consumption. Despite mean arterial pressure being higher in the 2K1C group when compared with controls (134±9 vs 97±10 mmHg, p=0.02), blood flow (7±1 vs 7±1 ml/min, p=0.823), glomerular filtration rate (0.42±0.1 vs 0.53±0.1 ml/min, p=0.45) and oxygen consumption (1.0±0.12 vs 0.6±0.1 mlO2/min, p=0.06) were not significantly different between 2K1C and control animalsThis preliminary data suggests that high blood pressure in 2K1C animals is sustaining normal renal perfusion, oxygen homeostasis and renal hemodynamics despite the reduction of arterial diameter.Support or Funding InformationBritish Heart Foundation funded research.
Discovery and Characterization of a Potent and Selective Inhibitor for Human Phosphoinositide‐3‐kinase γ
Authors: Meleza Cesar Alejandro , Chen Ada , Fournier Jeremy , Mailyan Artur , Mata Guillaume , Jeffrey Jenna , Banuelos Jesus , Zhao Xiaoning , Leleti Manmohan R. , Jin Lixia , Powers Jay P. , Walters Matthew J. , Young Stephen W. 
Published: April 2020

Phosphoinositide‐3‐kinases (PI3K) belong to the class 1 family of signal transducing enzymes comprised of PI3Kα, PI3Kβ, PI3Kδ and PI3Kγ. Together they are part of the PI3K‐Akt pathway acting as secondary messengers by catalyzing the phosphorylation of the inositol ring of phosphoinositides to regulate key cell functions such as proliferation, differentiation, and migration.Of the four isoforms, PI3Kγ has garnered attention for its expression on immune cells and its role in tumor progression, inflammation, and immune response. PI3Kγ activity has been correlated to cell proliferation and tumor growth in human pancreatic ductal adenocarcinoma (PDAC) and in hepato cellular carcinoma (HCC). Additionally, PI3Kγ inhibition is suggested to be a key switch within the tumor microenvironment between immune stimulation and suppression. This suggest that selective inhibition of PI3Kγ may be an effective anti‐cancer strategy.Previous attempts to selectively target PI3Kγ have proven largely unfruitful due to the poor selectivity profiles of many of the available inhibitors. We have developed a potent, reversible and selective small molecule PI3Kγ inhibitor. This inhibitor (compound 1) demonstrates activity against recombinant human PI3Kγ with sub nanomolar potency as determined by detection of ADP generation measured by luminescence. Additionally, compound 1 is highly selective over the rest of the class 1 PI3K family. Finally, compound 1 shows double digit nanomolar potency in MCP‐1 induced THP cells as measured by phosphorylation of AKT Ser473 using AlphaLisa and low triple digit nanomolar potency in whole blood as measured by flow cytometry.
The microtubule regulator ringer functions downstream from the RNA repair/splicing pathway to promote axon regeneration
Authors: Vargas Ernest John Monahan , Matamoros Andrew , Qiu Jingyun , Song Yuanquan 
Published: April 2020

Promoting axon regeneration in the central and peripheral nervous system is of clinical importance in neural injury and neurodegenerative diseases. Both pro‐ and antiregeneration factors are being identified. We previously reported that the Rtca mediated RNA repair/splicing pathway restricts axon regeneration by inhibiting the nonconventional splicing of Xbp1 mRNA under cellular stress. However, the downstream effectors remain unknown. Here, through transcriptome profiling, we show that the tubulin polymerization‐promoting protein (TPPP) ringmaker/ringer is dramatically increased in Rtca‐deficient Drosophila sensory neurons, which is dependent on Xbp1. Ringer is expressed in sensory neurons before and after injury, and is cell‐autonomously required for axon regeneration. While loss of ringer abolishes the regeneration enhancement in Rtca mutants, its overexpression is sufficient to promote regeneration both in the peripheral and central nervous system. Ringer maintains microtubule stability/dynamics with the microtubule‐associated protein futsch/MAP1B, which is also required for axon regeneration. Furthermore, Ringer lies downstream from and is negatively regulated by the microtubule‐associated deacetylase HDAC6, which functions as a regeneration inhibitor. Taken together, our findings suggest that ringer acts as a hub for microtubule regulators that relays cellular status information, such as cellular stress, to the integrity of microtubules in order to instruct neuroregeneration. Future studies include developing small‐molecule inhibitors for Rtca and further elucidating TPPP’s conserved function in regeneration.Support or Funding InformationY.S. is a recipient of the National Institute of Neurological Disorders and Stroke (NINDS) Pathway to Independence Award. This work was supported by an Intellectual and Developmental Disabilities Research Center New Program Development Award (CHOP/Penn), an NINDS K99/R00 award (R00NS088211), and a National Institutes of Health grant (1R01NS107392) to Y.S., and a Predoctoral Training Program in Developmental Biology (T32 HD083185) to E.J.M.V.