SRNT 26th ANNUAL MEETING new orleans maRCH 11-14, 2020 RAPID RESPONSE ABSTRACTS The peer-review process for SRNT’s annual meeting entails review by Society members of abstract submissions. Criteria for acceptance/rejection are based upon methodological rigor and not the funding resource or research findings. The views expressed by conference presenters are the author’s own and do not necessarily represent that of SRNT. 2020 Paper Sessions immediately prior to placement in a 20” x 9” x 18” open chamber for measurement of somatic withdrawal signs. Guide canulae were surgically implanted prior to any nicotine exposure. Results: Nicotine rats infused with vehicle into the IPN showed elevated RAPID SESSION 1: PRE-CLINICAL somatic withdrawal signs independent of route of nicotine exposure, suggesting that the procedures and surgery did not confound observation of withdrawal measures. Local IPN infusion of αCTXMII significantly attenuated withdrawal as measured by a reduction in somatic signs compared to nicotine withdrawn rats infused with vehicle in the IPN. Conclusions: The observation of this novel finding using two independent models of nicotine withdrawal suggests that α3β2* nAChR and/or α6β2* nAChR contribute to POD26-1 this measure of nicotine dependence and may be effective targets to promote nicotine CHANGE IN FREE BASE NICOTINE DELIVERY AS FUNCTION OF cessation in individuals who vape or smoke. This work was funded in part by the Virginia FLAVOR AND E-CIG BATTERY POWER OUTPUT Commonwealth University School of Medicine, Massey Cancer Center and NIH grants DA042749 and CA016059. Vinit V. Gholap, M.S.1, Leon Kosmider, PharmD, Ph.D.2, Matthew S. Halquist, Ph.D.1. 1 2 VCU School of Pharmacy, Richmond, VA, USA, Medical University of Silesia, Sos- FUNDING: Federal; Academic Institution nowiec, Poland. Introduction: Recent studies have shown nicotine salts, which yield protonated nicotine, gave higher plasma nicotine concentrations than free base (unprotonated) nicotine. Although the reason behind it is not well understood, it is important to classify e-liquids POD26-3 based on their free base or protonated nicotine yield in pre and post vaporization. Here, we report a study establishing correlation using variables of flavor and battery A ROLE FOR SECONDHAND SMOKE IN THE EPIGENETIC power. Methods: An aerosol generating model was developed using a syringe pump REGULATION OF HEPATIC GENES AND LIVER DISEASE based vaping machine. The model was validated using CORESTA guidelines (N°81). DEVELOPMENT Control e-liquids of five pHs (4-11), unflavored, with nicotine concentration 50mg/mL in PG:VG (70:30 v/v) were used for establishing a standard correlation between pre Hannah Blumenfeld, Ahmad Besaratinia, Stella Tommasi. University of Southern Cal- and post vaporization yield of free base nicotine. Using n=15 puffs at power settings ifornia, Department of Preventive Medicine, Los Angeles, CA, USA. (90W, 203°C) and 0.2Ω coil, nicotine was collected using a Cambridge filter. Impact Significance:Accumulating evidence shows that exposure to environmental pollutants, of flavor and battery power on free base nicotine yield correlation was tested using including secondhand smoke (SHS), contributes to the development of chronic fatty mango flavor and two different battery power settings. Pre and post vaporization free liver disease through generation of reactive oxygen species (ROS) and increased base nicotine determination was carried out using a Henderson Hasselbalch method burden of oxidative stress. Imbalances in the redox state of the cells are known to cause of the dilution approach. Results: The model passed CORESTA guidelines’ criteria alterations in the patterns of 5-hydroxymethylcytocine (5hmC), the oxidative product with standard square puff profile, puff flow 18.32+0.72 mL/sec, puff duration 3.00sec of 5-methylcytosine (5mC), and histone modifications, thus affecting the epigenetic and puff volume 55.16+0.3mL. Filter entrapment efficiency of nicotine was found to reprogramming and transcriptional regulation of genes involved in lipid metabolism be 97.88+2.32%. Control e-liquids showed positive correlation (Pearson r=0.994,) and/or oxidative stress response. Aberrant expression of these genes can induce between pre and post vaporization yield of free base nicotine. Mango flavor was found hepatic steatosis and exacerbate liver injury, thus promoting progression to advanced to deviate from the standard correlation and showed lower free base nicotine yield in forms of chronic liver disease. Methods: To investigate the impact of SHS on the liver post vaporization (94.77% pre vs 88.83+4.12% post). Higher power settings (120W, epigenome, we analyzed the relative gene expression of key epigenetic regulators, 303°C) further decreased free base nicotine yield in post vaporization (94.77% pre vs including DNA methyltransferases (DNMTs), ten-eleven translocation (TET) proteins, 80.97+4.15%, t-test, p=0.03). Conclusions: The mango flavor e-liquid was found to and histone deacetylases (HDACs) in the liver of mice subchronically exposed to SHS, lower post vaporization free base nicotine yield. Similarly, increase in battery power using quantitative real-time PCR. Furthermore, we quantified the global levels of over settings found to decrease the post vaporization free base nicotine yield. Future study eighty histone marks in SHS-treated mice and controls by mass spectrometry. Induction will be carried out with more flavors, nicotine salts and additional battery power settings. of oxidative stress was also assessed using a glutathione fluorescent detection protocol. The study is supported by Virginia Youth Tobacco Projects Small Funding Statement: Results: We observed significant SHS-induced changes in the relative expression of Grants Program for research funding and the Central Virginia Center on Drug Abuse Tet3, a major dioxygenase that catalyzes the oxidation of 5mC to 5hmC. Similarly, we Research (5P30DA033934-05). detected aberrant expression of Dnmt1 and Dnmt3a, and Hdac1, in the SHS-exposed mice relative to controls. Dysregulation of Hdac1 is consistent with changes in acetylation FUNDING: Federal; Nonprofit grant funding entity observed at lysine residues in position K27 and K36, which is often associated with gene activation. Depletion of glutathione (P<0.05), a biomarker of oxidative stress and a primary mechanism of ROS-induced toxicity in liver cells, was also confirmed in SHS-exposed mice. Conclusion: Our findings show that SHS can influence key POD26-2 components of the epigenetic machinery, which are involved in hepatic gene regulation, thus suggesting a role for SHS in the development of chronic liver disease. ALPHA CONOTOXIN MII-SENSITIVE NICOTINIC ACETYLCHOLINE RECEPTORS IN THE IPN SUPPORT SOMATIC WITHDRAWAL IN FUNDING: Federal; State LONG EVANS RATS Darlene H. Brunzell, PhD1, Janee Hayles1, Leah Aclin1, Jennifer Lee1, J. Michael McIntosh2, Paul Whiteaker3. 1Virginia Commonwealth University, Richmond, VA, USA, 2University of Utah, Salt Lake City, UT, USA, 3Barrow Neurological Institute, Phoenix, POD26-4 AZ, USA. VAPING ASSOCIATED ALTERATIONS IN THE LUNG LIPIDOME Significance: α conotoxin MII (αCTXMII)-sensitive receptors include the α6β2* and α3β2* Arunava Ghosh, Ph.D, James E. Keating, Raymond D. Coakley, Brandie M. Ehrmann, nicotinic acetylcholine receptors (nAChRs; *indicates possible assembly with α4 and Neil E. Alexis, Robert Tarran. University of North Carolina at Chapel Hill, Chapel Hill, β3). The α6β2* nAChR are enriched in the mesolimbic dopamine pathway where they NC, USA. support nicotine reinforcement and reward as well as in the interpeduncular nucleus (IPN) where they are co-expressed with α3β2* nAChRs. The IPN has been implicated Rationale: E-cigarette or vaping product use-associated lung injury (EVALI) is in regulation of nicotine withdrawal. The purpose of these studies was to locally and characterized by diffuse pneumonia, lipid laden macrophages, shortness of breath and selectively antagonize αCTXMII-sensitive nAChRs within the IPN to determine their gastrointestinal symptoms. Of note, altered lipids are associated with several types of contributions to nicotine withdrawal behavior. Methods: Rats were tested during lung disease including chronic obstructive pulmonary disease, pneumonia and lung spontaneous withdrawal 24 hr following removal of implanted minipump (6 mg/kg/day cancer. Since the underlying etiology is poorly understood, we hypothesized that lipids subcutaneous nicotine or 0.9% saline vehicle over 2 weeks) or 24 hr following removal and lipid-associated surfactant proteins may be altered in e-cigarette user’s lung. of Nicotine (25 µg/mL nicotine in 2% saccharin solution) or 2% saccharin alone in their Methods: Research bronchoscopy was performed on healthy subjects to collect homecages for 7+ weeks of exposure. All orally dosed rats received 1 hour/day access to bronchoalveolar lavage (BAL) samples from the three groups, namely non- water with overnight access to water in place of nicotine & saccharin prior to withdrawal smokers, smokers and e-cigarette users (vapers). Lipidome composition in the measures. Under both conditions, nicotine exposed rats showed significantly greater BAL fluid samples were analysed using gas chromatography-mass spectrometry somatic withdrawal than vehicle controls. Gently restrained rats received IPN infusion (GC-MS). We also measured the levels of surfactant proteins (SP) by Western of sterile ACSF (vehicle) or 10 pmol αCTXMII in ACSF through implanted guide canula