Poster abstracts

Title	Authors	Affiliations	Abstract
Therapeutic effect of novel EP2 receptor antagonist on neuroinflammation in mouse model of Alzheimer’s disease.	Avijit Banik, Radhika Amaradhi, Wenyi Wang, Nick Varvel, Ray Dingledine and Thota Ganesh	Department of Pharmacology and Chemical Biology, School of Medicine, Emory University. Atlanta, GA 30322	Prostanoid receptor signaling is found to be an important pathway inducing neuroinflammation in brain in injury, aging and neurodegenerative disorders (NDD). Cyclooxygenase-2 (COX-2) enzyme, a mediator for Prostaglandin synthesis, is induced in Alzheimer’s disease (AD) brain. Adverse cardiovascular effects of COX-2 inhibitors in the inflammatory conditions has shifted the focus on prostaglandin pathways, downstream of COX-2. Our recent studies showed the evidence of adverse role of prostanoid receptors in neuroinflammation after status epilepticus suggesting their involvement in AD pathogenesis. Thus, in this study, we examined anti-inflammatory effect of prostagnadin receptor (EP2) antagonist in AD brain. In this study, we identified a potent and selective EP2 antagonist, TG11-77.HCl, by its potency, selectivity and anti-inflammatory properties. Then we examined its effect of chronic treatment in the 5XFAD transgenic mouse model of AD. The mice were treated with TG11-77.HCl 100mg/kg/daily in drinking water and control mice were given vehicle drinking water from 3 months to 5 months of their age. Mice were also administered with a low dose (0.5 mg/kg) of lipopolysaccharide (LPS) once a week to induce a chronic level of brain inflammation in AD brain. There was no adverse effect of Tg11-77.HCl found on the behavioral scoring and body weight in these mice. Complete blood count (CBC) analysis revealed an inflammatory effect of LPS in WBC, RBC and platelet distribution, but it was not altered by TG11-77.HCl treatment. The drug treatment showed beneficial effects in brain reporting mRNA level of proinflammatory mediators (IL-1β, TNFα, IL-6, CCl2, CXCL10) and glial markers (IBA1, GFAP, CD11b, S110B) were significantly reduced by TG11-77.HCl treatment. When compared between sex, females always showed higher magnitude of inflammation in brain compared to males upon LPS injection and as expected, the drug was more effective in females compared to males. The overall number of amyloid plaques or area covered by them was not altered after the treatment. Overall, the EP2 antagonist could dampen the neuroinflammation in 5X-FAD brain without altering amyloid pathology when there was elevated levels of proinflammatory cytokines and Astro-glial loads after LPS insult, suggesting this drug could more efficiently address the neuroinflammation in later stage of the disease or associated with secondary infections.
Does lower energy expenditure increase reward-related cortical/brain activity and reward pursuit?	Mariane Bacelar, Boris Cheval, Matthieu P. Boisgontier, Robyn Feiss, Victoria Zona, Matthew W. Miller	Auburn University, University of Geneva, University of British Columbia, KU Leuven	Physical inactivity is a poorly understood and worldwide health problem. A recent theory contends that this pandemic may be explained by brain processes inherited from evolution. These processes would make us perceive low energy expenditure as a reward. However, direct empirical evidence testing the rewarding value of behaviors minimizing energetic cost is lacking. If this theory is correct, opportunities to earn rewards associated with low energy expenditure should elicit greater reward pursuit, as characterized by faster behavioral responses and increased response preparatory neural activity than rewards associated with higher energy expenditure (Hypothesis 1); and rewards associated with low energy expenditure should trigger greater reward-related neural activity (Hypothesis 2). We tested these hypotheses using an incentive delay task. In each trial of this task, participants attempted to quickly respond to a target (a square on a computer screen), then received reward feedback and retrieved a token or no reward feedback and no token. Half of the trials required participants to increase energy expenditure by standing up to retrieve the token, whereas the other half required participants to minimize energy expenditure by remaining seated to retrieve the token. Neural activity was recorded with EEG throughout the task. Response preparatory neural activity was assessed using the contingent negative variation (CNV) component preceding responses to targets, and reward-related neural activity was assessed using the reward positivity (RewP) component time-locked to feedback onset. Response times were also indexed. Dependent t-tests showed no differences in response time, CNV, or RewP between standing up vs. remaining seated trials, thus suggesting that energy expenditure had no influence on these outcomes. These findings do not support the theory that minimizing energy expenditure is rewarding. Future studies should investigate the factors that likely interact with the rewarding value of energy cost minimization such as cognitive resources or recent exercise history.
Identification of MAPT neuronal cis-regulatory elements	J. Nicholas Cochran, M. Natalie Davis, Mark Mackiewicz, Shelby Lauzon, Sarah K Strange, Christopher Partridge, Brian Roberts and Richard M. Myers	HudsonAlpha Institute for Biotechnology, Huntsville, AL, USA	Background: MAPT is associated genetically with multiple neurodegenerative diseases and codes for tau protein. Understanding the cis-regulatory elements associated with MAPT expression would inform on genetic determinants of disease risk, but the technology to comprehensively assess cis-regulatory elements has only been recently developed. Methods: We nominated candidate cis-regulatory elements for MAPT by assessing chromatin marks and Capture-C, and performed follow-up with luciferase assays and dCas9-KRAB inhibition. Results: We assessed nine candidate cis-regulatory elements nominated by chromatin marks by luciferase assays in cultures of inhibitory neurons. Three elements exhibited significant activity, and inhibition of two of these elements in situ using dCas9-KRAB resulted in reduced expression of MAPT. We also identified candidate cis-regulatory elements in proximity to the MAPT promoter by Capture-C in inhibitory neurons in triplicate at restriction fragment resolution. Conclusions: Expression of MAPT is likely to depend on recruitment of nearby cis-regulatory elements. Future studies should further address the function of these cis-regulatory elements, including how genetic variation in these elements may contribute to disease risk.
Differential sensitivity to novel stimuli and d-amphetamine following adolescent methylmercury exposure	Dalisa R. Kendricks, M. Chris Newland	Auburn University	Methylmercury (MeHg) is an environmental neurotoxicant known to induce alterations in dopamine-mediated behavior such as motor activity, reinforcement sensitivity, behavioral flexibility, response inhibition, and choice. Development of dopamine systems plays a key role in the manifestation of behavior related to attention-deficit/hyperactivity disorder (ADHD), including alterations in sustained attention and short-term memory. Alterations in attention and memory systems have been observed in children exposed to MeHg in utero following maternal seafood consumption. Because adolescence is also a key time for development of dopamine systems, the current study modeled potential changes in sustained attention and memory following exposure to MeHg throughout adolescence using a mouse model. Mice were exposed to 0, 0.3, or 3 ppm MeHg in drinking water throughout adolescence and trained in a visual signal detection task in adulthood. Sustained attention and memory were probed with novel tactile and visual stimuli. Behavior for all animals, across MeHg exposure groups, was impaired significantly by the tactile stimulus. However, animals exposed to 3 ppm were less impaired by the visual stimulus than the 0 and 0.3 ppm animals. In order to determine the characteristics of the visual stimulus, animals were injected with 0.3 mg/kg d-amphetamine and probed with the visual stimulus. Acute d-amphetamine improved memory in the presence of the visual distractor for the 3 ppm group but not for the 0 or 0.3 ppm groups. This differential sensitivity supports alteration in dopamine neurotransmission following developmental MeHg exposure and is in line with observations of behavioral inflexibility following developmental MeHg exposure.
Response inhibition to physical inactivity stimuli in an affective go/no-go task	Boris Cheval 1,2 , Marcos Daou 3, Daniel A.R. Cabral 4, Mariane F.B. Bacelar 4, Juliana O. Parma 4, Cyril Forestier 5, Dan Orsholits 6, David Sander 1,2, , Matthieu P. Boisgontie 7, Matthew W. Miller 4,8	1 Swiss Center for Affective Sciences, University of Geneva, Geneva, Switzerland, 2 Laboratory for the Study of Emotion Elicitation and Expression (E3Lab), Department of Psychology, University of Geneva, Geneva, Switzerland, 3 Department of Kinesiology, Coastal Carolina University, USA, 4 School of Kinesiology, Auburn University, USA, 5 Univ. Grenoble Alpes, SENS laboratory, Grenoble, France,6 Swiss NCCR “LIVES – Overcoming Vulnerability: Life Course Perspectives”, University of Geneva, Switzerland,7 Vancouver Community College,8 Center for Neuroscience, Auburn University, USA	Recent evidence suggests humans have an automatic attraction to effort minimization. Yet, how this attraction is associated with response inhibition is still unclear. Here, we used affective go/no-go tasks to capture inhibitory control in response to stimuli depicting physical activity versus physical inactivity in 59 healthy young individuals. Higher commission errors (i.e., failure to refrain a response to a “no-go” stimulus) were used as a measure for inhibitory control, whereas slower reaction times to go toward a “go” stimulus were used as an index of attentional bias. Based on the energetic cost minimization theory, we hypothesized that participants would exhibit higher commission errors and show slower reactions when responding to physical inactivity, compared with physical activity, stimuli. As expected, mixed effects models showed that participants exhibited higher commission errors (odds ratio = 1.59, 95%CI = 1.18 to 2.16, p = .003) and showed slower reaction times (b = 26.4, 95%CI = 11.0 to 41.7, p = .001) when responding to stimuli depicting physical inactivity compared with physical activity. These results suggest that physical inactivity stimuli might attract attention and require high response inhibition. This study lends support for the hypothesis that an attraction to effort minimization might affect attention and inhibitory function processes in the presence of stimuli related to this minimization.
Angiotensin II Impairs Neuro-Glial Unit Nrf2 Activity in Central Cardioregulatory Nuclei of Spontaneously Hypertensive Rats	Francesca E. Mowry, Sarah C. Peaden, and Vinicia C. Biancardi	Anatomy, Physiology & Pharmacology, College of Veterinary Medicine, Auburn University Center for Neuroscience Initiative, Auburn University	Within central cardioregulatory nuclei, Angiotensin II (AngII) directly influences neurons and glia through type I receptors (AT1R), yet the cell type-specific pathway(s) by which central AngII actions propagate hypertension remain unclear. We hypothesized that AngII dysregulates Nrf2-based antioxidant defenses, promoting astrogliosis and neuroinflammation. 12-13 week-old spontaneously hypertensive rats (SHRs) and normotensive Wistar Kyoto rats (WKYs) were used to examine protein (immunofluorescence [IF]) and mRNA (RT-PCR) alterations in the hypothalamic paraventricular nucleus (PVN) and rostral ventrolateral medulla (RVLM). Confocal imaging of IF assays were analyzed for single-cell and subcellular (nuclear and non-nuclear [cytosolic]) Nrf2 colocalization with NeuN (neurons) or GFAP (astrocyte). Interleukin (IL)-6 and tumor necrosis factor (TNF)-α IF densities were elevated in SHR nuclei (PVN: IL-6:+19.9±5; TNF-α:+147.2±23%; RVLM: IL-6:+52.2±7; TNF-α:+119.6±20% vs. WKY), indicating a pro-inflammatory cytokine profile consistent with astrocyte reactivity. Nrf2 density was reduced in SHR neurons (PVN: cell:-43.3±4.0, nucleus:-33.3±6.7, cytosol:-63.9±6.8%; RVLM: cell:-31.0±2.9, nucleus:-55.2±6.8, cytosol:-46.7±5.9% vs. WKY) and astrocytes (PVN: cell:-55.2±5.0, nucleus:-71.9±4.0, cytosol:-54.1±5.7%; RVLM: cell:-50.5±6.4, nucleus:-72.9±3.7, cytosol:-61.8±4.5% vs. WKY). SHR PVN punches showed increased mRNA levels of the Nrf2-repressor Keap1 (2.1±0.2), alongside reductions in Nrf2-regulated antioxidant genes, Gpx1 (0.7±0.1) and NQO1 (0.5±0.1-fold change), versus WKYs. To determine the relative contribution of AngII, SHRs were treated with Losartan (AT1R inhibitor; 20mg/kg/day; gavage; SHRLos). After four weeks, mean arterial pressure (tail-cuff volume-pressure system) was normalized in SHRLos (SHR:154.9±1.7; SHRLos:103.6±2.4mmHg), and altered Nrf2 densities were reversed in neurons (PVN: cell:-2.8±6.8, nucleus:-5.8±6.5, cytosol:+4.6±13.9%; RVLM: cell:+1.3±6.8, nucleus:+2.8±13.6, cytosol:-2.8±13.9% vs. WKY) and astrocytes (PVN: cell:-3.9±9.0, nucleus:+1.2±11.9, cytosol -4.1±10.2%; RVLM: cell:+5.1±18.5; nucleus:-5.6±15.6; cytosol:-7.1±19.2% vs. WKY). Furthermore, AT1R inhibition abolished the increased levels of GFAP density within the PVN of SHRs (WKY:15.9±0.6, SHR:26.4±0.7, SHRLos:15.3±0.6% area). Taken together, these findings suggest that AngII impairs Nrf2-based antioxidant defense systems and promotes pro-inflammatory astrocyte reactivity within cardioregulatory nuclei of hypertensive animals.
Time-frequency Spectral Analysis of Scalp EEG signals using Empirical Mode Decomposition	Jingyi Zheng, Liangming Li, Arne Ekstrom, Fushing Hsieh.	Auburn University, Department of Mathematics and Statistics, University of Arizona, Department of Psychology, University of California, Davis, Department of Statistics.	Electroencephalography (EEG) is an electrophysiological monitoring method to record electrical activity of the brain from different electrodes placed on scalp. In this talk, we propose an effective data-driven processing pipeline to first clean, decompose the scalp EEG signals, and perform time-frequency analysis using Hilbert-Huang transform, and further build machine learning classifiers. Specifically, we propose to decompose a EEG signal into a set of Intrinsic Mode Functions (IMFs) using a data-driven method named Empirical Mode Decomposition (EMD). For each IMF, we obtain the instantaneous frequency and amplitude through Hilbert transform. Then we propose a new metric to measure the frequency component of each IMF and further select the IMF to represent certain brain wave. We also propose a metric to measure the averaged amplitude of certain brain wave. Using the proposed metrics, we can test some cognitive hypothesis, as well as build machine learning classifiers. Specifically, we first train a between task classifier using Random Forest to successfully classify four tasks with 99.12% accuracy in average, and further build within task classifiers to classify body/eye movements for each task with at least 94.47% accuracy.
Breaching Barriers in Glioblastoma: Comparison of Currently Available Proteasome Inhibitors	Andrey Maksimenko, Jennifer Fields, Will Colley, Steven N. Fiering, Robert D. Arnold, Alexei F. Kisselev	Dept. of Drug Discovery & Development, Harrison School of Pharmacy, Auburn University	Glioblastoma multiforme (GBM) is a fast-growing glioma that develops from star-shaped glial cells (astrocytes and oligodendrocytes) that support the health of the nerve cells within the brain. These tumours are highly cancerous with a high cell proliferation level. New therapeutic strategies are being developed worldwide to fight against deadly GBM, which has a median survival time of just 14 months. Proteasome inhibition has shown promising results in cancers such as myeloma. However, this form of therapy has also shown positive results in brain tumours in the way of elevated apoptosis. A head-to-head comparison of clinically available proteasome inhibitors (PIs) shows that in the clinically relevant setting, only the co-inhibition of beta-2 with beta-5 activity achieves significant functional proteasome inhibition and cytotoxicity. The selective inhibition of beta-5 subunit is sufficient to induce cytotoxicity in PI-sensitive, but not in PI-resistant GBM, and beta-5/beta-2 co-inhibition is the most cytotoxic in PI-resistant GBM.
Cerebellar lobulus simplex involvement in spatial decision making and prefrontal-hippocampal gamma coherence	Yu Liu, Samuel S. McAfee, Roy V. Sillitoe, Detlef H. Heck,	Department of Anatomy and Neurobiology, University of Tennessee Health Science Center, 855 Monroe Ave., Memphis, TN 38163, USA Department of Pathology and Immunology, Program in Developmental Biology, Baylor College of Medicine, Jan and Dan Duncan Neurological Research Institute of Texas Children’s Hospital, Houston, TX 77030, USA Department of Neuroscience, Program in Developmental Biology, Baylor College of Medicine, Jan and Dan Duncan Neurological Research Institute of Texas Children’s Hospital, Houston, TX 77030, USA St. Jude Children’s Research Hospital, Division of Translational Imaging Research, Memphis, TN 38105, USA	Cerebellar cognitive function requires cerebro-cerebellar communication via a network of reciprocal connections. Neuronal activity in individual cerebral cortical areas can me modulated by manipulating neuronal activity at specific cerebellar sites and vice versa, but it remains unclear how this exchange of activity contributes to cognitive functions. We addressed this question by conducting triple-site recordings of single unit Purkinje cell (PC) spike activity in cerebellar cortex lobulus simplex (LS) or Crus I and of local field potential (LFP) activity in the medial prefrontal cortex (mPFC) and dorsal hippocampal CA1 area (dCA1) in awake, head fixed and in freely moving mice. Our rationale for targeting these three areas is based on their joint involvement in spatial navigation and spatial working memory (SWM) (1). During the decision-making process in a SWM task oscillations in the mPFC and dCA1 show a brief increase in coherence, which is believed to be essential for the decision making process (2). Our results show that Purkinje cells in LS and Crus I represent preferred phases and phase differences between oscillations in the mPFC and dCA1 (3), suggesting a possible involvement of the cerebellum in the phase alignment of mPFC-dCA1 oscillations. We show that loss of cerebellar function impairs decision related mPFC-dCA1 coherence and SWM performance. Additional results showing that photoactivation of PCs in LS of healthy mice increases the number of incorrect SWM decisions support a causal involvement of cerebellar LS in the decision-making process. We propose that the cerebellum contribution to SWM decision-making consists in the coordination of coherence, and hence of the functional connectivity between the mPFC and dCA1. 1. J. M. Lefort et al., Cerebellum 14, 59-62 (2015). 2. J. A. Gordon, Curr Opin Neurobiol 21, 486-491 (2011). 3. S. McAfee et al., Cell reports 27, 2328–2334 (2019).
The effect of distinct settings and types of exercise images on the automatic processing of non-exercisers	Juliana O. Parma, Victoria Zona, Mariane Bacelar, Daniel Cabral, Matthew W. Miller	Performance and Exercise Psychophysiology Lab (PEPLab), School of Kinesiology, Auburn University	Physical inactivity is one of the main risk factors for non-communicable diseases. Thus, mass-reach campaigns are usually developed to encourage sedentary people to undertake an active life. These campaigns often use exercise images, and recent work suggests the type of exercise (gym-based vs. sport-based) and setting of exercise (indoors vs. outdoors) depicted in the images affects reflective affective evaluations of the images. Yet, other recent work indicates reflective and automatic affective evaluations of exercise images differ. Given the important role of automatic processing in the adoption of active behavior, we investigated non-exercisers’ automatic evaluations of exercise images as a function of the exercise type and setting depicted in the images. We used an affective oddball paradigm in which participants were presented with series of negative, neutral, or positive images among which one of four types of exercise images was presented: indoor/gym-type; indoor/sport-type; outdoor/gym-type; and outdoor/sport-type. We obtained participants’ reflective ratings of the images, and the amplitude of the P3b component of participants’ electroencephalography (EEG)-derived event-related potentials evoked by the exercise images served to assess participants’ automatic evaluations. Results revealed that exercise images were reflectively rated as neutral/positive, and this rating did not depend on image setting or type. Regarding automatic evaluations of exercise images, those set indoors and those depicting gym-type exercise elicited greater P3b amplitude than those set outdoors and those depicting sport-type exercise, respectively. Yet, these effects were not moderated by whether the exercise images were presented in a series of negative, neutral, or positive images, suggesting that differences in how the exercise images were automatically evaluated were not related to affect, but just to the magnitude of cognitive processing. Thus, exercise images may be automatically processed to differing extents based on the setting and type of exercise depicted in the images..
Behavioral tasks that assess recognition memory are useful tools to assess cognitive deficits in preclinical models.	Tyler Stone1,2*, Alexus Williams1*, Luke Watson1,3, Dominique Williams1 and Catrina Robinson1,3	1 Department of Neurology, Medical University of South Carolina, Charleston, South Carolina, 29425 USA, 2 College of Charleston, Charleston, South Carolina, 29425 USA, 3 Molecular and Cellular Biology & Pathobiology Program, Medical University of South Carolina, Charleston, South Carolina, 29425 USA	Obesity is associated with cognitive impairment, specifically deficits in executive functions such as working memory and problem solving. Studies demonstrate that high-fat diet (HFD) mice, a model of obesity, have deficits in working memory using tasks such as the novel object recognition task (NOR). In NOR, normal memory is evident by spending more time exploring the novel object following a familiarization phase. Given that mice use their vibrissae to explore and navigate the environment, it is not known whether deficits in such tasks can be attributed to HFD-induced sensory deficits. Furthermore, it is not known whether HFD mice exhibit deficits with problem solving. Our hypothesis is the HFD-induced obesity leads to non-sensory dependent cognitive deficits in both working memory and problem solving. To determine the potential impact of sensory deficits on working memory tasks, we modified the NOR, by replacing objects with sandpaper grits, to create the novel tactile recognition task (NTR). Our data demonstrates that the NTR task is whisker dependent task evident by the inability to identify the novel sandpaper grit following bilateral whisker removal in mice on a standard diet (STD). Furthermore, the inability to determine the novel sandpaper grit in HFD mice compared with the STD, suggests that HFD mice have impairments in tactile recognition memory. To determine the potential impact of HFD on whisker sensitivity, mice were subjected to the corner task. HFD mice do not exhibit any deficits in the corner task compared with the STD mice. To evaluate problem solving, STD and HFD mice were subjected to the activity box, which consists of two compartments, an open field and an enclosed dark box, connected by an escape door. Obstacles were placed in front of the escape door to create the problem-solving task. Our data demonstrates that the HFD mice have increased escape latencies compared to the STD mice. Collectively, our data demonstrates that HFD leads to deficits in both working memory and problem solving, which are not due to sensory deficits.
Using Systems Genetics to Better Model Age-related Macular Degeneration (AMD)	T.J. Hollingsworth, Anand Swaroop, Emily Chew, David Ashbrook, Robert W. Williams, Monica M. Jablonski	Hollingsworth and Jablonski – University of Tennessee Health Science Center, Department of Ophthalmology, Hamilton Eye Institute, Memphis, TN Williams and Ashbrook – University of Tennessee Health Science Center, Department of Genetics, Genomics, and Bioinformatics, Memphis, TN Swaroop and Chew – National Eye Institute, National Institutes of Health, Bethesda, MD	Generating animal models of AMD has been an arduous task with limited results due to the heterogenous nature of the disease including numerous known genetic components along with several environmental factors such as age, smoking, and diet. Our hypothesis is that a model containing multiple single-nucleotide polymorphisms (SNPs) in genes associated with AMD will provide a phenotype more similar to that seen in humans compared to models with single genes altered. Thus, we are applying a systems genetics approach toward modeling AMD using the BXD family of mouse strains. Using genes known to have an association with AMD diagnosis, we assessed for the following criteria: the genes must be self-regulating (cis-eQTL regulation); they must have a differential expression between both C57BL/6J (B6, normal) and DBA/2J (D2, mutant) mouse strains (the parent strains of the BXD family); and they must have SNPs present in the D2 strain. Post-analyses, BXD mouse strains were assessed for strains possessing the most D2 haplotypes for the genes-of-interest ApolipoproteinE (Apoe), Complement Factors 3 (C3), B (Cfb), H (Cfh) and I (Cfi), Collagen Type VIII Alpha 1 Chain (Col8a1), Retinaldehyde Binding Protein 1 (Rlbp1), Serine protease HTRA1 (Htra1), Syndecan 2 (Sdc2), Vascular Endothelial Growth Factor A (Vegfa), and Vascular Endothelial Growth Factor Receptor 2 (Kdr), and B6 haplotypes for the Tyrp1 and Gpnmb genes to remove any confounding due to pigmentary dispersion glaucoma. The strains selected will be aged to 18 months, and anatomical and functional tests will be performed including optical coherence tomography, electroretinogram, funduscopy and optokinetic nystagmus tests. Preliminary data from BXD mice containing D2 haplotypes for both Cfh and Cfi reveal the combined insult from these genetic variations produce an AMD-like histopathological phenotype that is more pronounced than either gene alone, including larger and more numerous basolaminar deposits, increased RPE vacuoles and undigested phagosomes.
Factors influencing CNS distribution of intrathecally-administered antisense oligonucleotides	Brynna Wilken-Resman, Michelle E. Pizzo, Robert G. Thorne	University of Wisconsin-Madison, University of Wisconsin-Madison, Denali Therapeutics	Antisense oligonucleotide (ASO) therapies for certain CNS indications have advanced in recent years, including FDA approval for an ASO therapy for spinal muscular atrophy and clinical trials for other neurological disorders. ASOs are ~7 kDa, single-stranded DNA oligomers and are attractive therapeutic candidates for diseases caused by known genetic abnormalities because they can interact with target RNA to modify protein production. ASOs are unable to cross the blood-brain barrier on their own, so they are administered centrally, typically achieved by intrathecal administration. Here, we investigated ASO transport and distribution in the CNS following intrathecal administration, focusing on the effects of different ASO chemistries and two transport mechanisms: diffusion in the extracellular spaces and convection/dispersion in perivascular spaces surrounding the cerebral vasculature. Three fluorescently-labeled ASOs differing in chemical modification and sequence were used for the following experiments. ASO1 was a fully phosphorothioated oligonucleotide. ASO2 was partially phosphorothioated and contained a 2’MOE modification. ASO3 had neutral phosphorodiamidate morpholino modifications. Integrative optical imaging was used to measure the diffusion coefficients and apparent hydrodynamic sizes of the ASOs. Intrathecal administration of ASOs was conducted in rats and the distribution was visualized using ex vivo fluorescence and confocal microscopy. Signal corresponding to perivascular ASO was quantified using a custom Fiji/ImageJ program. ASOs were found to have apparent hydrodynamic diameters in the range of 3-4 nm. Ex vivo fluorescence imaging after intrathecal infusion in rats revealed a substantially more limited diffusion gradient and perivascular distribution for ASO1 compared to ASO2. The difference in perivascular signal between the two ASOs was significant when quantified, and was particularly apparent in the dorsal cortex as well as in subcortical brain regions such as the striatum and hippocampus. Each ASO has a different distribution pattern based on their characteristics. ASOs may experience additional sources of hindrance aside from molecular weight (i.e., secondary structure, binding, or electrostatic interactions) that could limit their distribution in the brain via diffusion and perivascular access. Thorough understanding of the mechanisms underlying ASO distribution in the brain and the factors limiting its delivery could lead to strategies to enhance the CNS delivery of ASOs.
Differential utilization of visual and proprioceptive information for postural control stability in individuals with autism spectrum disorder (ASD)	Zheng Wang, Ph.D.1, Jiaqi Chen 2, Emily In 3, Walker S. McKinney, B.S. 4-6, Zhigang Li, Ph.D.2, Matthew W. Mosconi, Ph.D.4-6	1 Department of Occupational Therapy, University of Florida, Gainesville, FL 32611, USA 2 Department of Biostatistics, University of Florida, Gainesville, FL 32611, USA Department of 3 Microbiology and Cell Science, University of Florida, Gainesville, FL 32611, USA 4 Schiefelbusch Institute for Life Span Studies, University of Kansas, Lawrence, KS 66045 5 Clinical Child Psychology Program, University of Kansas, Lawrence, KS 66045, USA 6 Kansas Center for Autism Research and Training (K-CART), University of Kansas Medical School, Overland Park, KS 66213, USA	Sensorimotor impairments are common in autism spectrum disorder (ASD), although the extent to which sensory processing issues affect motor control and movement coordination in individuals with ASD is not well understood. Previous studies have suggested that individuals with ASD show increased postural sway variability in stances during which either visual or proprioceptive information is occluded. No known study to date has examined the interactive effect of visual and proprioceptive inputs on postural stability in individuals with ASD. The aim of this ongoing research is to fill this critical knowledge gap by quantifying individuals’ postural sway variability during tasks with and without the availability of visual and proprioceptive information. Twenty-eight school-aged children with ASD and 11 age-, sex- and IQ- matched healthy controls participated in tests of static stance on an AMTI force platform. During the assessment, a pair of lightweight tendon vibrators (TVs) was attached on participants’ Achilles tendon of the ankle joints. When turned on, both TVs vibrate at a high frequency of 80Hz to create a transient proprioceptive illusion of lengthened Achilles tendon of both legs. Participants completed tests with and without the TVs (i.e., TVon vs. TVoff) and lights on and off (LTon vs. LToff). Postural sway variability was assessed using the center of pressure (COP) trajectory length and COP standard deviation in the anterior-posterior (COPAP) and mediolateral (COPML) directions. Both individuals with ASD and healthy controls showed increased COP trajectory length and COPAP standard deviation during TVon conditions regardless of the light condition. The COP trajectory length of patients, relative to controls, showed a larger increase during TVon compared to TVoff conditions. Individuals with ASD showed increased COPAP and COPML standard deviation compared to controls across all task conditions. Proprioceptive information serves as the primary sensory source for gross motor activities including postural control. Our results suggest that postural instability in individuals with ASD and healthy controls is more severely impacted by proprioceptive illusions compared to perturbations in visual feedback. Although both individuals with ASD and TD controls were affected by the proprioceptive illusion, postural stability was more strongly impacted in ASD suggesting a heightened dependency on proprioceptive information during standing. These findings inform a more mechanistic view of postural issues in ASD indicating that over-reliance on proprioceptive feedback may interfere with stability in patients.
Intranasal insulin treatment improves functional stroke outcome in mice	Crystal Smith1,2, Stacy Nguyen1,3, and Catrina Sims-Robinson1,2*	1Department of Neurology, Medical University of South Carolina, Charleston, South Carolina, 29425 USA, 2Department of Neurosciences, Medical University of South Carolina, Charleston, SC 29425 USA, 3College of Charleston, Charleston, South Carolina, 29425 USA	Strokes pose both a physical and financial burden on patients, it has been estimated by the CDC that strokes cost the United States $34 billion dollars due to treatment and inability to work. The road to rehabilitation after stroke can be a long strenuous process many recovering stroke patients require physical and cognitive rehabilitation. Often times rehabilitation focus is placed on trying to restore physical abilities while the cognitive deficits are not addressed with the same urgency. Many stroke patients not only require rehabilitation but are also diagnosed with hyperinsulinemia; characterized as elevated levels of insulin circulating in the blood resulting from reduced insulin transport from the periphery to the brain, causing an insulin deficiency. Brian insulin promotes neuroplasticity, synaptogenesis, has anti-inflammatory, anti-thrombotic, vasodilatory, anti-apoptotic properties, and is involved in cognition. The benefits of brain insulin give way to numerous research avenues that promote stroke recovery. We hypothesize that intranasal insulin treatment will improve functional recovery following an ischemic stroke in a mouse model of hyperinsulinemia. To explore our current hypothesis ischemic stroke is induced using a 30-minute middle cerebral artery occlusion (MCAO) in a high-fat diet induced and standard diet induced male mouse model (n>11). The animals are assessed on the following criteria; survival, weight, neurological severity score, motor and cognitive function after receiving either intranasal saline (0.9%) or intranasal insulin (1.75U). Preliminary results indicate high-fat diet-induced hyperinsulinemia leads increased adverse neurological deficits and functional recovery following ischemic stroke. Furthermore, our preliminary studies indicate that intranasal insulin may have advantageous benefits improving functional recovery from stroke in the high-fat mice.
Hyperinsulinemia induced brain microvessel insulin resistance correlates with reduced insulin transport	Luke S. Watson, Crystal J. Smith, Alexus S. Williams, Catrina Sims-Robinson	1 Department of Neurology, Medical University of South Carolina, Charleston, South Carolina 2 Molecular and Cellular Biology and Pathobiology Program, Medical University of South Carolina, Charleston, South Carolina 3 Department, Medical University of South Carolina, Charleston, South Carolina	During Alzheimer’s disease and stroke, two of the most common age related neurodegenerative disorders, patients present with systemic hyperinsulinemia. Systemic hyperinsulinemia is associated with reduced CNS insulin levels, however the mechanisms underlying this are not well understood. Insulin is fundamental in the brain for neuroplasticity, and has anti-apoptotic, -inflammatory, -thrombotic, and -vasodilatory properties. Insulin must be transported across the blood brain barrier (BBB). Some have suggested this occurs via insulin receptor-mediated transcytosis. Thus, the purpose of the following studies are to determine the impact of hyperinsulinemia on insulin transport across the BBB. In the current study, at four weeks of age, B6 mice were placed on either a standard diet (STD) or a high-fat diet (HFD) for 6 and 24 weeks. Hippocampal microvessels were extracted and the levels of serine-phosphorylated insulin receptor substrate, a docking protein that can inhibit insulin receptor function, were measured. Furthermore, these microvessels were stimulated with insulin to determine the ability to activate downstream insulin signaling. We observed evidence of insulin resistance in HFD microvessels and impaired insulin signaling. To explore whether these alterations impacts insulin transport, we utilized primary brain endothelial cells, a key component of the BBB, and exposed the cells to hyperinsulinemic conditions to evaluate insulin resistance, signaling, and transport. Our data demonstrates that insulin resistance and impaired insulin signaling correlates with reduced transport. Future studies will seek to validate these findings in more complex BBB systems.
Effect of administration route and AAV serotype for treatment of feline GM1 gangliosidosis	Amanda L. Gross, Heather L. Gray-Edwards, Miguel Sena-Esteves, Douglas R. Martin	Scott-Ritchey Research Center, Department of Anatomy, Physiology, and Pharmacology, College of Veterinary Medicine, Auburn University, Alabama Department of Radiology, Department of Neurology, Horae Gene Therapy Center, University of Massachusetts Medical School, Massachusetts	GM1 gangliosidosis is a fatal neurodegenerative disease caused by a deficiency of lysosomal β-galactosidase (βgal). Cerebrospinal fluid (CSF) administration of adeno-associated viral (AAV) therapy is hypothesized to be an effective method for treating neurodegenerative diseases. In this study, we evaluated two serotypes (AAV9 and AAVrh10) using CSF delivery via the cisterna magna (CM). Additionally, we compared these results to intravenous administration of AAV9. All treatment cohorts received 1.5e13 vector genomes/kg body weight at 1.8 ± 0.5 months of age. Untreated GM1 animals survived 8.0 ± 0.6 months while treated animals lived significantly longer. Clinical assessments included neurological exams, CSF biomarkers, and 7T magnetic resonance imaging (MRI) and spectroscopy (MRS). Postmortem analysis included βgal and virus distribution. Neurological abnormalities, which in untreated GM1 animals lead to the inability to stand by 8 months of age, were delayed but not halted in both CM cohorts. The IV treated cohort had mild neurological symptoms, similar to those seen early in disease stages, but no further impairments. MRI revealed delayed progression of neurodegeneration in both CM cohorts and preservation of brain architecture in the GM1+AAV9 IV cohort. Glycerophosphocholine and phosphoscholine, a MRS biomarker that increases with loss of myelin integrity, showed no correction in the GM1+AAVrh10 cohort, correction only in the cerebellum of the GM1+AAV9 CM cohort, and correction in several brain locations of the IV cohort. In the CM cohorts, βgal activity was restored in the cerebellum and spinal cord but did not penetrate deep brain structures (such as thalamus). The GM1+AAV9 IV cohorts had increased βgal activity throughout the CNS. All cohorts had some degree of βgal restoration in peripheral tissues. Using a similar vector backbone and same total dose, this study demonstrates AAV efficacy in all treatment cohorts and suggests that IV gene therapy is most effective in the feline model.
How do presynapses capture the resources they need from the axon?	Matthew E. White, Faith Taylor, Madison Lowery, Michael W. Gramlich	Auburn University, College of Science and Mathematics	The presynapses that release neurotransmitter, in order to communicate with the post-synapse, recycle most of their resources. However, some resources, such as synaptic vesicles, are shared between presynapses along the same axon in a process called inter-synaptic vesicle exchange (ISVE). This ISVE requires that resources leave one presynapse transport along the axon and be captured by another presynapse. Moreover, ISVE has been shown respond to neuronal activity suggesting that it supports cognition, but little is known about the molecular mechanics for how this resource sharing process works. We focus on the mechanics of how synaptic vesicles are captured from the axon into a presynapse. We focus on three parameters that have previously be shown to regulate ISVE: activity, myosin-V, the actin cytoskeleton. We show that all three components are required for presynapses to capture vesicles.
Modifying glutamatergic neurotransmission rectifies synaptic plasticity and memory deficits in a 3xTg Alzheimer’s disease model	Jeremiah C. Pfitzer, Priyanka D. Pinky, Irfan A. Qureshi, Robert M. Berman, Vishnu Suppiramaniam, Miranda N. Reed	Department of Drug Discovery and Development, Auburn University, Auburn, AL, USA. Biohaven Pharmaceuticals, Inc., New Haven, Connecticut, USA. Center for Neuroscience Initiative, Auburn University, Auburn, AL, USA.	Alzheimer’s disease (AD) poses a public health crisis, and there is an urgent need for effective treatments and disease-modifying therapies. Glutamate dysfunction is a core feature of AD, and glutamate is a validated target for treating AD. We tested a novel, rationally designed 3rd generation tripeptide prodrug of the glutamate modulator, riluzole, that offers improved bioavailability, safety, and dosing. It is currently being examined in a phase 2 trial for patients with mild-to-moderate AD (NCT03701399). In this preclinical study, we examined the effects of this drug on synaptic function using the triple transgenic (3xTg) AD model. 3xTg mice received the riluzole prodrug (25 mg/kg/day) from 5 -8 months of age. At 8 months of age, drug-treated 3xTg mice were compared to vehicle-treated 3xTg and nontransgenic controls for alterations in synaptic plasticity using acute hippocampal slices. Basal synaptic transmission was reduced by 50-60% in vehicle-treated 3xTg slices compared to controls, and treatment partially attenuated these basal synaptic transmission deficits. To determine if differences in basal synaptic transmission were related to presynaptic modifications, the probability of neurotransmitter release was examined by measuring paired pulse facilitation (PPF). PPF was significantly reduced in vehicle-treated 3xTg slices, indicating an increase in presynaptic release probability. Drug treatment restored PPF in 3xTg mice to that of controls. The amount of readily releasable pool (RRP) of synaptic vesicles was also significantly reduced in 3xTg mice, indicating increased vesicle docking. This treatment significantly increased RRP in 3xTg mice. We next measured long-term potentiation (LTP), a cellular correlate of learning and memory. LTP was reduced by 25% in vehicle-treated 3xTg mice compared to controls. Notably, drug treatment completely restored the synaptic plasticity deficits observed in 3xTg mice. These results suggest that this riluzole prodrug restores synaptic deficits in 3xTg mice and highlight its potential as a novel therapy for AD.
Western Diet-Induced Obesity Disrupts the Diurnal Rhythmicity of Peripheral Metabolism and Inflammation as well as Hippocampal Core Clock Gene Expression in a Male Mouse Model.	Lauren N. Woodie, Robert M. Johnson, Bulbul Ahmed, Savanah Fowler, William Haynes, Beatriz Carmona, Miranda Reed, Vishnu Suppiramaniam, Michael W. Greene	Department of Nutrition, Auburn University, Auburn, AL Center for Neuroscience, Auburn University, Auburn, AL Department of Biological Sciences, Auburn University, Auburn, AL Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University, Auburn, AL Boshell Diabetes and Metabolic Disease Research Program, Auburn University, Auburn, AL	Western diet (WD) feeding disrupts core clock gene expression in peripheral tissues and contributes to WD-induced metabolic disease. The hippocampus, the mammalian center for memory, is also sensitive to WD feeding, but whether the WD disrupts its core clock is unknown. To this end, male mice were maintained on a WD for 16 weeks and diurnal metabolism, gene expression and memory were assessed. WD-induced obesity dampened the diurnal rhythms of whole-body metabolism and hepatic gene expression, but did not disrupt diurnal expression of hypothalamic Bmal1, Npas2 and Per2. However, all measured core clock genes were disrupted in the hippocampus after WD feeding and the expression pattern of genes implicated in Alzheimer’s disease and synaptic function were altered. Finally, WD feeding disrupted hippocampal memory in a task- and time-dependent fashion. Our results implicate WD-induced alterations in the rhythmicity of hippocampal gene expression in the etiology of diet-induced memory deficits.
Elucidation of the mechanisms of learning and memory deficits in adolescent offspring due to prenatal cannabinoid exposure	Priyanka D. Pinky 1, Jenna Bloemer 1, Sharay E. Setti 1, Ryan T. Heslin 1, Yifeng Du 1, Warren D. Smith 1, Alexander Dityatev 2,3, Muralikrishnan Dhanasekaran 1,2, Miranda N. Reed 1,2, Vishnu Suppiramaniam 1,2	1 Auburn Univ Harrison School of Pharmacy, Auburn, AL, USA; 2Center for Neuroscience Initiatives (CNSi) Auburn University, Auburn, AL, USA; 3German Center for Neurodegenerative Disease, Magdeburg, Germany	Cannabis is currently the most used illicit drug in the USA. 6-8% of pregnant women use cannabis to reduce morning sickness and improve appetite. Cannabis can cross blood placental barrier to reach the fetus causing enduring alterations in synaptic circuitry responsible for cognition and behavior. The aim of our study is to investigate the mechanism of learning and memory deficits due to prenatal cannabinoid exposure (PCE) in adolescent offspring. An osmotic pump filled with either vehicle or the cannabinoid receptor agonist WIN55,212-2 (2 mg/kg/day) was implanted subcutaneously in gestational Day-3 to deliver the drug until the pups were born. An open field test revealed no significant difference of PCE on general locomotor activity of the offspring. Hippocampus based task contextual fear conditioning test revealed a significantly reduced freezing behavior in the PCE animals. Hippocampus based spatial memory test Morris water maze revealed the preference for the platform quadrant in control animals while the PCE animals did not show any preference during the probe trial. Electrophysiological experiments on acute hippocampal slices showed impairment in synaptic plasticity parameters including long term potentiation and long-term depression. These impairments were dependent on the change of glutamate receptor subtype NMDA channel activity. An increase in cannabinoid receptor type 1(CB1) expression followed by reduced neural cell adhesion molecule (NCAM) and its active form polysialylated NCAM (PSA-NCAM) expression have been observed in western blot analysis. Later, LTP in the PCE group has shown to be improved by using a PSA-mimetic compound. Based on our findings we hypothesize that, observed behavioral deficits were due to altered NMDA mediated glutamatergic neurotransmission due to reductions in the signaling through NCAM/PSA NCAM and its downstream protein molecules. Understanding this specific NCAM/PSA NCAM-NMDA interdependent signaling cascade will lead to an identification of a specific therapeutic target to improve cannabinoid mediated memory deficits.
The potential threat of party pills on addiction and Parkinson’s disease	Mohammed Almaghrabi; Mohammed Majrashi; Ahmad Almalki; Jack Deruiter; C.Randall Clark; Tim Moore; Muralikrishnan Dhanasekaran	Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University Department of Medicinal Chemistry, College Of Pharmacy, Taibah University, KSA	Different botanical derived substances or synthetic addictive drugs have been “misused” and/or “abused” for centuries around the world. To overcome the abuse by these substances, strict legal laws are constituted globally. However, novel and drugs with chemical structures similar to illegal psychoactive drugs substances (with a slight structural change) are manufactured in undercover laboratories to have the same or augmented psychostimulatory effects. The objective of the current study is to understand the consequence of abusing piperazine compounds that may lead to Parkinsonism. The concern about the rate of Parkinson’s disease is rising alarmingly worldwide due to many factors. Meanwhile, misuse of synthetic piperazine compounds also increased, particularly among the young generation. Consequently, chronic use of stimulants or drugs of abuse can alter the neurotransmission, induce neurotoxicity and thereby act as catalytic factors for the emergence of dopaminergic neurodegenerative disorders. Subsequently, this requires high priority research to ascertain the neurotoxic profile of these compounds. Therefore, our goal is to examine the neurotoxicity profile of the new and novel psychoactive substances (piperazine derivatives) and determine their mechanisms of neurotoxicity. Our hypothesis is that chronic abuse of piperazine products can have a significant influence on the monoaminergic neurotransmitters which may produce Parkinsonism. Our specific aims are to determine the neurotoxic potential of novel piperazine derivatives. To achieve this goal, we will utilize cell proliferation assay to evaluate the differential neurotoxicity of synthetic piperazine derivatives on the N27 dopaminergic neurons and detect the mechanisms of neurotoxicity. Piperazine derivatives induced oxidative stress and mitochondrial dysfunction leading to dopaminergic neurotoxicity. Thus, there is a strong possibility for piperazine derivatives can induce monoaminergic neuronal insult and increase the risk for dopaminergic neurodegeneration leading to parkinsonian symptoms.
The contribution of endogenous tau to alpha-synuclein inclusion formation	Lindsay E. Stoyka, Casey Mahoney, Andrew Arrant, Ashwin Narayanan, David G. Standaert, Erik Roberson, Laura A. Volpicelli-Daley	University of Alabama at Birmingham, Center for Neurodegeneration and Experimental Therapeutics, Medical Scientist Training Program	Parkinson’s disease (PD) is characterized by loss of dopaminergic neurons within the substantia nigra pars compacta and intracellular inclusions composed mostly of alpha-synuclein (α-syn). Symptoms classically include tremor, rigidity, bradykinesia, and postural instability. In addition to extrapyramidal motor effects, patients experience a variety of non-motor symptoms, with up to 80% of patients eventually developing cognitive changes, termed Parkinson’s disease dementia. A negative correlation between the density of α-syn pathology and the mini-mental state examination (MMSE) suggests that α-syn pathology contributes to cognitive deficits. In addition to α-syn, GWAS studies also implicate the microtubule-binding protein tau as a risk factor for PD, though the exact role of endogenous tau in PD is unknown. The presence of co-morbid neurofibrillary tangles composed of tau is more common in PD dementia than PD, suggesting that tau also contributes to the cognitive phenotypes. We show that bilateral injections of α-syn fibrils into the striatum results in robust α-synuclein inclusion formation in the cortex and amygdala, particularly in excitatory neurons. Fibril injected mice show defects in a social dominance behavioral task associated with prefrontal cortex function. They also show defects in fear conditioning, which is associated with amygdala function. Reduction of tau lowers fibril-seeded α-syn inclusion burden in the amygdala and cortex. Additionally, we show, using super-resolution microscopy, that tau and α-syn colocalize at the presynaptic terminal in the cortex and, using a proximal ligation assay, tau and α-syn interact on a molecular level in neurons. Further work will explore if the decrease in inclusions rescues cognitive phenotypes in tube test and fear conditioning. Together, these observations suggest that an interaction of α-syn and tau in the cortex contributes to PD-related phenotypes, and that tau reduction may be an important therapeutic approach for alleviating non-motor deficits of PD.
Enhanced bioavailability of Boswellic acid by Piper longum in Rabbits: A computational and pharmacokinetic analysis	Mansour Alturki, K. Reeta Vijayaranib, Mohammed Majrashi, Ayaka Fujihashi, N. Kirubakaranb, Forrest Smith, Muralikrishnan Dhanasekaran	Department of Pharmaceutics, Periyar College of Pharmaceutical Sciences, Sundar Nagar, Tiruchirappalli – 620 021, India, Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University, AL 36849, USA, Department of Pharmaceutical Chemistry, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam 34212, Saudi Arabia, Department of Pharmacology, Faculty of Medicine, University of Jeddah, Jeddah, 23881, Saudi Arabia	Chronic inflammation substantially increases the risk of morbidity and mortality. Hence, novel therapeutic approaches are currently being designed to discover new treatment avenues or improve the pharmacokinetic and pharmacodynamic effects of existing synthetic/natural drugs. Boswellic acids are well-known natural products which have the ability to decrease inflammation effectively without severe adverse effects. However, the use of Boswellic acids is impeded due to its poor pharmacodynamic properties. Pharmacokinetics strategies that can increase the absorption and distribution of Boswellic acids can lead to a safe and effective pharmacological product that can be used prophylactically and therapeutically to decrease inflammation. In this study, we investigated the effect of Piper longum on the bioavailability of Boswellic acid in a valid animal model. Additionally, we also explored the computational pharmacodynamic interaction of Piper longum and Boswellic acid. Piper longum dose-dependently increased the bioavailability of Boswellic acid. Based on our drug-based computational studies, the cytochrome P450 mediated mechanism can be involved in the increased bioavailability, and these studies also confirm that these drugs can be administered orally for effective therapeutic efficacy. Thus, computational aided drug design with validated pharmacological experiments supports the use of Piper longum with Boswellic acid as the future novel therapeutic avenue for treating various types of inflammation.
Asymmetry of Spatial-temporal Mutual Priming and Effect from Reading Habit	Yi Chen	University of Alabama, College of Education, Educational psychology (Educational Neuroscience)	This study explored the relationship between asymmetry of spatial-temporal mutual priming and the effect of reading habit. Two experiments were conducted to subjects from both Chinese Mainland and Taiwan. Experiment 1 primed the spatial concept by time concept. The between-subjects factor is the source of the subjects (from mainland or Taiwan), within-group factor is the directions of presenting spatial question (vertical or horizontal). The reaction time to time question were recorded as the dependent variable. The main effects and interactions were significant, which indicated that reading direction would affect the dominant direction of spatial theory priming temporal theory, even under the same language background. Procedure in the second experiment is basically as same as Experiment 1, except that time question was ahead of spatial question, which was used to start the spatial concept by time concept. Neither the main effect nor the interaction was significant. Comparing the second experiment with the first indicate the spatial-temporal mutual priming and had asymmetry.
A Regulatory Intersection of Dopamine and miRNA Transport in C. elegans Parkinson’s Disease Models	J. Brucker Nourse Jr., Nathan A. Moniz, Ryan A. Tuckey, Adam C. Holzhauer, Kim A. Caldwell & Guy A. Caldwell	University of Alabama, Caldwell Lab	A novel therapeutic target of Parkinson’s disease (PD) called TNK2, a non-receptor tyrosine kinase, has come out of a recent whole exome sequencing study involving familial PD patients. Based on the location in the protein sequence of the 4 SNP’s discovered, we postulate that TNK2 cannot be negatively regulated by the E3 ubiquitin ligase NEDD4 and its prolonged activity is deleterious for dopaminergic neurons. TNK2 plays a role in preventing the endocytosis of the dopamine reuptake transporter (DAT) in mammals. Caenorhabditis elegans (C. elegans) possess an ortholog of TNK2, known as SID-3, which is involved in the internalization of dsRNA into the cytoplasm, possibly through the conserved dsRNA transporter SID-1. Bringing these findings together, we hypothesize that TNK2/SID-3 co-regulates the import of dopamine and dsRNA, and that this mechanism can be targeted for a therapeutic benefit in PD patients. To test the hypothesis, we modulated the activity of SID-3 in C. elegans with a knockout mutation and pharmacologically with Aim-100 (TNK2 inhibitor) and NAB2 (NEDD4 activator) in assays involving 6-hydroxydopamine (6-OHDA) exposures, a-synuclein (a-syn) overexpression in the dopaminergic neurons, and RNAi studies targeting GFP. Our results find that the dopaminergic neurons of sid-3(ok973) mutants are protected from 6-OHDA and a-syn toxicity. Treatments with Aim-100 and NAB2 protected the dopaminergic neurons of C. elegans from both insults. NAB2-induced protection was abolished in mutants for the NEDD4 homolog wwp-1(gk372). Aim-100, but not NAB2, prevented RNAi. Our studies indicate that SID-3 functionally behaves as the mammalian homolog to regulate the internalization of DAT, and the loss of SID-3 activity is neuroprotective and impedes RNAi. These findings demonstrate that SID-3 can be manipulated in vivo with two different drugs to achieve neuroprotection and RNAi inhibition.
Motivations for Cannabis Use Are Associated with Psychotic Symptomatology	Regan A. Moss, Julio A. Yanes, Jennifer L. Robinson	Department of Psychology, Center for Neuroscience Initiative	There is well-documented use of, and dependence on, psychoactive substances among people with prodromal syndromes and psychotic diagnoses. In particular, cannabis use is prevalent in schizophrenia patients. Strikingly, lifetime use of cannabis is twice as high amongst schizophrenics as compared to the general population. Importantly, however, use of illicit drugs, such as cannabis, is detrimental to people with schizophrenia. Specifically, cannabis use can exacerbate psychotic symptomology and lead to more detrimental outcomes. Though the co-occurrence of cannabis use and psychosis has been well-documented, few studies have sought to provide clarification regarding underlying motivations to use cannabis and their implications for subsequent psychopathology. In this study, we aimed to (i) assess cannabis use motivations among college students who completed the Marijuana Motives Measure (MMM), a five-factor model which includes conformity, coping, enhancement, expansion, and social use motives, and (ii) explore associations between motives and scores on the Prodromal Questionnaire-Brief (PQB) Version, which measures prodromal symptoms. Among 288 participants, confirmatory factor analysis (CFA) provided some support for the MMM five-factor model (X2 = 635.537, df = 220, p < 0.001) with modest fit indices (SRMR = 0.091 , CFI = 0.902, RMSEA = 0.082). Subsequent regression analyses using the five factors as predictors showed associations between cannabis motives and prodromal symptoms. Specifically, expansion (p = 0.008) was positively associated with prodromal symptoms, while social motives, enhancement, conformity, and coping were not. The finding that expansion is positively associated with cannabis use among individuals in our study with higher scores on the PQB may indicate that individuals with greater endorsement of prodromal symptoms may use in order to reduce negative symptomology that cannabinoid effects counteract. In fact, the effects of cannabis that are commonly sought-after are described in the expansion factor (i.e., perceptual and cognitive enhancement). Thus, these results also support the need for future studies to better understand the pathway between prodromal symptoms, such as those proceeding schizophrenia, and cannabis usage. In other words, do people use cannabis for the expanded experiences and as a consequence, screen higher for prodromal symptoms, or do individuals with higher scores of prodromal symptomology seek cannabis, such that the expanded experiences may deplete negative symptomology.
Association of depressive symptoms and suicidal ideation with impulsive choice: Findings from nationally representative survey	Kaniz A. Tanni, Kimberly B. Garza	Health Outcomes Research and Policy, Harrison School of Pharmacy, Auburn University	An increasing body of evidence suggests that serotonin dysfunction can be a trigger for depressive disorders, thoughts of self-harm, as well as impulsivity. We conducted a cross-sectional survey of a nationally representative sample of US adults over age 18 (n= 478) via Qualtrics to explore risk factors for depressive symptoms (DS) and suicidal ideation (SI). Our central hypothesis was that respondents with greater impulsivity are more likely to have self-reported DS and SI. Respondents having a total score >10 the PHQ9 scale for depression and a score >0 on the 9th item of the PHQ9 were identified as having DS and SI respectively. Impulsivity was measured using scores from both the well-established Barratt Impulsiveness Scale (BIS-11) and the newly introduced Three-option Adaptive Discount rate (ToAD) measure. Chi-squared tests were performed to determine association of DS or SI with categorical demographic variables. Logistic regression was performed to determine whether DS and SI are associated with impulsivity scores, controlling for demographics. Race, ethnicity, income level, and marital status were found to be significantly associated with SI (p= 0.037, 0.003, 0.030, 0.0002 respectively). For depressive symptoms, significant associations were found with race, gender, income level and marital status (p= 0.008, <0.0001, 0.0008 and <0.0001 respectively). One point increase in the log of discount rate on ToAD scale (higher score represents higher impulsivity) was found to increase the likelihood of having SI and DS by 12% (OR = 1.12 [95% CI, 1.01 – 1.26]) and 13% (OR=1.13 [95% CI, 1.01-1.27]) respectively. Similar odds of suicidal ideation (OR= 1.05 [95% CI, 1.03-1.06]) and depressive symptoms (OR = 1.05 [95% CI, 1.04- 1.07]) were found when impulsivity was measured by BIS-11 scale. Findings from this study will have potential implications in designing future behavioral interventions.
Identifying Epigenetic Modulators of Neuronal Resilience in the C. elegans Dopaminergic System	Anthony L. Gaeta, Luke E. McKay, Candice M. Keogh, Kim A. Caldwell, and Guy A. Caldwell	The University of Alabama	By studying individuals resilient to the stressors that normally severely affect the susceptible majority, novel treatments may be discovered and developed to combat neurodegenerative diseases like Parkinson’s disease (PD). In the case of PD, the most implicated genetic stressor is the misfolding-prone protein α-synuclein (α-syn), which induces cellular dysfunction through various mechanisms that ultimately lead to the death of dopamine (DA) neurons which account for the symptoms experienced by PD patients. We have generated a model of PD by overexpressing α-syn in the DA neurons of the nematode roundworm Caenorhabditis elegans (C. elegans), which exhibits progressive, age-dependent dopaminergic neurodegeneration as seen in humans. Although a vast majority of individuals that constitute this model worm strain are susceptible to the stress of α-syn, there exists a small minority of individuals whose DA neurons never degenerate, even in the face of α-syn. It is our hope that these individuals hold the key to discovering new genes, small non-coding RNAs, and/or epigenetic modifications that may lead to more effective treatments or preventative measures for PD. In order to facilitate this goal, we have devised of a way to enrich for resilient populations of worms, in the context of our PD model, by selecting α-syn-expressing worms based on a behavioral readout of DA neuron health, the Basal Slowing Response (BSR). Resilient populations of worms generated from this BSR selection method have resulted in populations of worms that show significantly less dopaminergic neurodegeneration than their control counterparts. In order to discover what genetic or epigenetic mechanisms these resilient populations may be using to confer protection to α-syn, we isolated RNA from both control and resilient populations of worms and employed RNA sequencing to find gene transcripts which were differentially expressed between the two populations.
How do presynapses capture the resources they need from the axon?	Matthew E. White, Faith Taylor, Madison J. Lowery, Michael W. Gramlich	Auburn University	The presynapses that release neurotransmitter, in order to communicate with the post-synapse, recycle most of their resources. However, some resources, such as synaptic vesicles, are shared between presynapses along the same axon in a process called inter-synaptic vesicle exchange (ISVE). This ISVE requires that resources leave one presynapse transport along the axon and be captured by another presynapse. Moreover, ISVE has been shown respond to neuronal activity suggesting that is supports cognition, but little is known about the molecular mechanics for how this resource sharing process works. We focus on the mechanics of how synaptic vesicles are captured from the axon into a presynapse. We focus on three parameters that have previously be shown to regulate ISVE: activity, myosin-V, the actin cytoskeleton. We show that all three components are required for presynapses to capture vesicles.
Loss of cerebellar Purkinje cell output increases gamma coherence between the cerebellum and prefrontal cortex.	Brittany L. Correia, Yu Liu, Angela P. Taylor, Roy V. Sillitoe, Detlef H. Heck	Department of Anatomy and Neurobiology, University of Tennessee Health Science Center	Long known for its role in motor control, it is increasingly clear that the cerebellum is involved in numerous cognitive behaviors. Though the neuronal mechanisms for the role of the cerebellum in cognition are still unclear, recent work in our lab suggests the cerebellum monitors and possibly coordinates the precise timing of neuronal oscillations in cerebral cortical areas. Specifically, we showed that Purkinje cells in right lobulus simplex (LS) of healthy mice represent the instantaneous phase and phase differences of oscillations in the medial prefrontal cortex (mPFC) and the dorsal hippocampus (dHC) (McAfee et al., 2019, Cell Reports, 27:2328). Here we measured neuronal oscillations in the cerebellar LS and the mPFC and dHC in healthy mice, while they were at rest in their home cage. We quantified the average magnitude of coherence of oscillations in this resting state and asked whether resting-state cerebrocerebellar coherence would be affected by loss of cerebellar function. Our mouse model of cerebellar deficit suffered from a genetically induced loss of Purkinje cell neurotransmission (White et al. 2014, J. Neurosci., (34)24:8231), resulting in ataxic motor deficits typical for cerebellar dysfunction. Resting-state coherence of gamma oscillations between LS and mPFC was significantly increased in ataxic mice relative to their healthy littermates. There was no difference between healthy and ataxic mice in resting-state coherence between LS and dHC, or between mPFC and dHC. Anatomical tracing studies in primates have suggested that the cerebrocerebellar connectivity is organized in closed loops, making a unidirectional influence of on structure on the other unlikely. It is thus currently unclear whether the cerebellum exerts control on mPFC oscillations to modulate coherence or vice versa, or whether both structures are jointly involved to reach an equilibrium baseline coherence. In future analyses we will apply granger causality analysis to address this question.
A systematic review of motor sequelae of acute and chronic use of methylphenidate	Adefunke O. DadeMatthews, Abby Brittain, Adelia Grabowsky, Kristina A. Neely	Auburn University, School of Kinesiology	Background: Methylphenidate (MPD) is a first-line pharmacological treatment for Attention-Deficit/Hyperactivity Disorder (ADHD). This systematic review explores the effects of MPD on movement in humans and other mammals. We focus on objective measurements of movement in combination with manipulation of MPD. Although cognitive and behavioral effects of MPD have been extensively studied, fewer studies have investigated movement. There are substantial differences in measurement techniques and statistical analyses across studies. Search strategy: The PRISMA checklist for systematic reviews was followed (Moher et al., 2009). The concepts of Methylphenidate and Motor activity along with synonyms, related terms, and subject headings were searched on 9/30/2019 in PubMed, PsycINFO, International Pharmaceutical Abstracts, and ProQuest Dissertations and Theses. The initial searches yielded 1819 results, 1202 remained after deduping. Titles and abstracts were examined against the inclusion and exclusion criteria, and 886 were found to be non-suitable. Evaluation of full-text articles is in progress. Inclusion criteria: Primary and peer-reviewed research including observational studies, cohort or cross-sectional design, in vivo studies, studies with measures of motor output, methylphenidate comparison to control, studies in English and those published before 10/1/2019. Exclusion criteria: Secondary and/or non-peer-reviewed research including books, reviews, systematic reviews, meta-analysis, editorials, commentaries, meeting abstracts, case studies, case and laboratory notes. Studies with simultaneous administration of more than one drug, without original data, or without a control. Discussion: Although this systematic review is in the screening and eligibility phase, it is clear that limited reports of quantitative and objective measures of movement in relation to MPD exist. This project will review the findings of included studies and provide tabulated summaries.
Differential Effects of Learning on Neuronal Activity in Mouse Insular Cortex	Stephanie M. Staszko (1,2), John D. Boughter(1), Max L. Fletcher(1)	(1)Department of Anatomy and Neurobiology, (2) Integrated Biomedical Sciences Program	Previous studies investigating the effects of learning on neuronal activity within the taste region of insular cortex (IC) have reported conflicting results. The primary explanation of these differences is the anatomical location of IC in the mouse brain, which makes reliably recording responses from IC neurons across days difficult. Our lab utilizes miniaturized, head-mounted microscopes (miniscopes) to image the calcium activity of neurons in awake, behaving animals. Importantly, this technology allows us to image the activity of the same population of neurons over multiple days to investigate the potential effects of learning on neuronal activity. In our paradigm, mice are trained to lick taste stimuli from a Davis Rig, which allows for the control of stimulus presentation as well as the recording of individual licks. Animals were tested in non-associative and associative learning paradigms. In non-associative learning, animals are presented with a taste panel over multiple days. As animals become familiar to the taste stimuli, they will increase their consumption of preferred stimuli (sweet) and decrease consumption of non-preferred stimuli (bitter), where animals learn to increase consumption of preferred tastes (sweet) and decrease consumption of non-preferred tastes (bitter). In the associative learning known as conditioned taste aversion (CTA), animals learn to avoid a taste after it has been paired with gastric malaise using the salt lithium chloride (LiCl). We have found populations of taste responsive and non-taste responsive neurons in IC. With non-associative learning, the activity of non-taste responsive neurons rapidly decreases. However, after CTA, the activity of this population increases, typically exceeding baseline levels of IC activity. We are currently interested in determining single cell changes in these cell populations following learning.
Exposure to an Environmental Contributor of Parkinson’s Disease: S. venezuelae impacts lifespan and mitochondrial dynamics in C. elegans	Jennifer L. Thies, Hanna Kim, Guy A. Caldwell, and Kim A. Caldwell	The University of Alabama, Tuscaloosa, Department of Biological Sciences	Parkinson’s Disease (PD) is characterized by the loss of dopaminergic (DA) neurons and the formation of protein inclusions that contain the α-synuclein (α-syn) protein. Overexpression of human α-syn in the eight DA neurons of C. elegans causes neurodegeneration in an age- and dose-dependent manner, similar to that observed in human pathology. Only 5-10% of PD cases have a direct genetic origin; however, exposure to herbicides, pesticides, and interaction with soil are all potential risk factors. A soil bacterium, Streptomyces venezuelae (S. ven), produces a secondary metabolite that causes age- and dose- dependent DA neurodegeneration in C. elegans; it also exacerbates α-syn-induced DA neurodegeneration. Initial studies from our lab determined that exposure to the S. ven metabolite caused oxidative stress and upregulation of reactive oxygen species (ROS). These studies identified the metabolite worked through the transcription factor daf-16 to activate sod-3. We also found that S. ven toxicity negatively impacts mitochondrial function, associated with increased mitochondrial fragmentation. To determine whether the metabolite impaired the longevity of worms, we subjected wild-type (WT) worms to lifespan assays. We discovered that exposure to the S. ven metabolite caused a decrease in C. elegans lifespan. However, further studies revealed metabolite exposure at lower concentrations, caused lifespan extension, suggesting a hormetic effect. Notably, daf-16 mutants displayed no significant differences between solvent control and metabolite at both high and low concentrations in lifespan assays, suggesting the hormetic response is daf-16 dependent. We additionally investigated the impacts of S.ven metabolite on the aging process of C. elegans mitochondrial fission and fusion mutants, drp-1 and fzo-1, respectively. We found exposure to S.ven increased lifespan in a drp-1 mutant background, suggesting that this cellular pathway, which we have shown to be upregulated in response to S.ven exposure, might be important for combating toxicants following chronic exposure.
Correlation of behavioral indicators of neurotoxicity with biomarkers of hepatotoxicity in mice co-administered with doxorubicin and cyclophosphamide	Julia M. Salamat, Kodye L. Abbott, Mohammed Majrashi, Mohammed Almaghrabi, Manoj Govindarajulu, Sindhu Ramesh, Kristina Gill, Eddie Fahoury, Natasha Narayanan, Darshini Desai, Rishi Nadar, Edwin McElroy, Timothy Moore, Muralikrishnan Dhanasekaran, Satyanarayana R. Pondugula	Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine, Auburn University Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University	Chronic exposure to chemotherapeutics can result in severe neurotoxicity and hepatotoxicity. A combination chemotherapy regimen of doxorubicin and cyclophosphamide is employed in treatment of several cancers including leukemia, lymphoma, and breast cancer. We therefore sought to determine whether co-administration of doxorubicin and cyclophosphamide results in neurotoxicity and/or hepatotoxicity. Male C57BL/6J mice received one intraperitoneal injection of saline or chemotherapeutics (doxorubicin-2mg/kg & cyclophosphamide-50mg/kg) per week for 4 weeks. After the treatment period, indicators of cognitive functions, liver histology and serum biomarkers of hepatotoxicity were assessed. Co-treatment of doxorubicin and cyclophosphamide induced severe cognitive impairment as seen in Y-maze, Water maze and Novel Object Recognition tests. In contrast, the chemotherapeutics did not alter markers for hepatotoxicity such as the serum levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP), bilirubin, albumin, globulin, or total protein. Similarly, coadministration of chemotherapeutics did not result in a noticeable change in liver histology. It was notable that the chemotherapeutics treatment resulted in a significant increase in serum levels of aspartate aminotransferase (AST). Elevated serum AST levels were also associated with increased serum creatinine kinase (CK) levels and decreased body weight after treatment with the chemotherapeutics, suggesting that elevated serum AST levels could be due to chemotherapeutics-induced muscle damage. Together, these results suggest that coadministration of doxorubicin and cyclophosphamide, induces neurotoxicity without inducing significant hepatotoxicity in mice. However, dose-dependent and time-dependent future studies are warranted to further confirm the above findings.
Association of salivary neurotrophins and insulin in childhood obesity	Vaithinathan Selvaraju, Ramesh Babu Jeganathan, Geetha Thangiah	Department of Nutrition, Dietetics, and Hospitality Management, Auburn University, Auburn, AL, USA	The global epidemiologic of metabolic disorder is associated with obesity, and it increases the risk of developing insulin resistance, diabetes, and cardiovascular disease. Obesity with other metabolic disorders also reported being associated with the onset of neurodegenerative disease. This study aimed to examine the level of the salivary BDNF, NGF, and insulin in normal weight, overweight and obese children, and its correlation with obesity measures. Forty normal weight (NW), twenty overweight (OW), and sixteen obese (OB) children aged 6-10 years participated in this study. Anthropometric measurements, blood pressure, and heart rate were recorded. The level of salivary BDNF, NGF, and insulin was measured using a human premixed magnetic Luminex assay kit. The expression of salivary BDNF, NGF, and insulin was significantly increased in OB compared to NW children. The systolic and diastolic blood pressure was also significantly increased in OB participants. BDNF, NGF, and insulin showed a positive association with the anthropometric measurement and among the parameters. In conclusion, our study suggests that a high level of salivary BDNF, NGF, insulin, and blood pressure are associated with obesity in children.
Antibody Gene Therapy for Neurological Infections	Henry J. Baker, Douglas R. Martin, Tatiana Samoylova, Amanda Gross, and Satheshkumar S. Panayampalli	Scott-Ritchey Research Center, College of Veterinary Medicine, Auburn University and the Center for Disease Control and Prevention, Atlanta, GA.	Circulating antibodies are excluded from the nervous system (CNS) by the blood-brain-barrier (BBB); therefore infections of the CNS are not protected by traditional vaccines. We developed antibody gene therapy using recombinant adeno-associated virus vectors that continuously deliver antibodies passed the BBB and prevent and treat infections of the nervous system for life, after a single administration. We demonstrated that antibody gene therapy produced neutralizing rabies antibodies systemically and in brain at concentrations 1000 fold higher than that considered by the World Health Organization to be protective.
Exploring the role of the mitochondrial unfolded protein response in a Parkinson’s Disease model	Corey W. Willicott, Guy A. Caldwell, Kim A. Caldwell	The University of Alabama	Parkinson’s disease (PD) is the second most common neurodegenerative disease that is characterized by the presence of misfolded α-synuclein in Lewy Body inclusions and the loss of dopaminergic neurons. Though causes of PD are unknown, there is evidence for mitochondrial dysfunction playing a role in the development of the disease. A strategy that mitochondria employ in response to stress is the mitochondrial unfolded protein response (UPRmt). Activation of this response in the presence of acute stressors promotes cell survival, however, chronic activation can be detrimental to the cell. α-Synuclein can interact with TOMM20, an outer mitochondrial membrane protein and form a mitochondrial import block. It is proposed that the increased UPRmt in dopaminergic neurons is the result of this interaction. We hypothesize that overexpression of TOMM-20 should alleviate this block and restore mitochondrial import, reducing cell death. Data from this experiment indicates that TOMM-20 overexpression, in fact, increases dopaminergic cell loss. Future work will be done to explain this result and elicit a possible mechanism for the observed increase in cell death.
Hematological markers detect cognitive impairment in rodent models tested with streptozotocin and chemotherapy	Caroline A. Jackson, S Pondugula,, M Majrashi, M Almaghrabi, K Abbott, M Govindarajulu, S Ramesh, K Gill, E Fahoury, N Narayanan, D Desai, R Nadar, E McElroy, T Moore, M Dhanasekaran	Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine, Department of Drug Discovery Development, Harrison School of Pharmacy, Auburn University	There are currently multiple factors that can contribute to cognitive impairment, including endogenous factors, such as hyperglycemia, & exogenous factors, such as medications and illicit drugs. Since many of the methods that indicate and measure cognitive decline are expensive and invasive, we attempted to determine if there are hematological markers that may predict cognitive impairment. Using rodent models of cognitive impairment, two different chemicals were administered to measure the presence of cognitive decline. This included a streptozotocin induced hyperglycemic model and chemotherapy (doxorubicin/cyclophosphamide) induced model. Hematological markers in the two rodent model groups were analyzed and compared to controls. In the models treated with chemotherapy, there was a noticeable increase in creatinine kinase, lactate dehydrogenase and aspartate aminotransferase (AST). In the models treated with streptozotocin, there was a significant increase in blood urea nitrogen (BUN), alkaline phosphatase (ALP), bilirubin, creatinine and glucose levels. However, both the chemotherapy and streptozotocin models exhibited elevated levels of triglycerides. Previously, human studies have shown a potential relationship between elevations in triglyceride levels and cognitive impairment. Similarly, our research using rodent models indicates that an increase in triglycerides correlates to cognitive impairment. Therefore, this suggests that the elevation of triglycerides in the blood could be a non-invasive procedure to aid in determining an increased risk for cognitive impairment. It is important to do further studies to determine the relationship between cognitive impairment and elevated triglyceride levels.
Identification of UPRmt Effectors Associated with a-syn Neuroprotection in a C. Elegans Model	Karolina Willicott, Rachel M. Bolden, Madeline R. Greene, A. Rose Davidson, Kim A. Caldwell, Guy A. Caldwell	The University of Alabama	Parkinson’s disease (PD) is characterized by neurodegeneration of the central nervous system, leading to impaired motor skills and bodily functions. Here, we look at two components of PD pathology: One is the protein a-synuclein that is found within Lewy Bodies and is often prone to misfolding and aggregation. The other is mitochondrial dysfunction via a dysregulated Unfolded Protein Response (UPRmt). Under non-stress conditions, the Activating Transcription Factor Associated with Stress (ATFS-1) is imported into the mitochondria and degraded. During acute stress, mitochondria employ the (UPRmt), that, with the help of ATFS-1, coordinates nuclear expression of chaperones and proteases, that translocate to the mitochondria and remove damaged or unfolded proteins. However, during a long-term genetic stressor such as a-synuclein, this system becomes dysregulated. A physical blockade is created when a-synuclein interacts with the outer mitochondrial membrane protein, TOMM-20. It is our hypothesis this blockade increases the UPRmt response because it prevents ATFS-1 and any chaperones from entering the mitochondria. These proteins build up in the cytosol and lead to a proteostatic imbalance. Our preliminary research showed that the loss-of-function mutant of ATFS-1 was neuroprotective. Thus, in our current study, we will take advantage of the ATFS-1 (lof) strain generated previously and use it to reveal an uncharacterized compensatory mechanism for UPRmt induction through a forward genetic screen with the roundworm C. elegans.
Pseudomonas aeruginosa elicits amyloid production and tau aggregation in the brain	Iva Durdanovic, Allison Scott, Dr. Mike T. Lin	Univeristy of South Alabama College of Medicine (Department of Physiology and Cell Biology)	The mortality rate in critically ill patients has improved significantly over the past 20 years, attributable to advanced ventilation strategies and hemodynamic support. As one of the main contributors to higher mortality rate and cognitive decline, Pseudomonas aeruginosa is an optimistic a bacterium known to cause nosocomial pneumonia in intensive care unit (ICU) patients, which induces cytotoxic amyloid production from endothelial cells and tau hyperphosphorylation. This neurotixicty can be tracked and observed through experiments including staining of the particular brain parts, where and Aβ oligomers are present as a result of their production in pulmonary endothelial cells. These oligomers create amyloid plaques and tangles in the brain that cause neurological disfunction during and after lung infection. The purpose of this project was confirming the connection between P. Aeuaginosa induced lung infection and production of Aβ-amyloids and Tau fibrils in brain. In our experiment, the staining was performed initially with intratracheal inoculation of PA103 and ∆PcRV (non-virulent) bacterium strains into mice expressing human Alzheimer’s precursor protein (hAPP) and in non-AD BL6 mice. Mice were then harvested for 5 weeks, and each week four sets of brains were obstained, sliced and fixed overnight. Following these steps, the staining was performed through DAB and Immunofluorescense stainings. The Primary and HRP/fluorescently labeled secondary antibodies were carefully chosen and the imaging was performed following conclusions of all the experiments. The hAPP and a non-AD brains both showed the presence of plaques and tangles, which supported our original hypothesis.
Combined Etodolac and α-tocopherol As a Novel Therapeutic Approach for Alzheimer Disease Treatment	Ihab M. Abdallah, Amal Kaddoumi	Drug Discovery and Development, Harrison School of Pharmacy, Auburn University,	Alzheimer’s disease (AD) is a complex neurodegenerative disorder with multiple dysfunctional pathways. Therefore, advanced treatment strategy that simultaneously targets multiple brain cell types and disease pathways could be advantageous for effective intervention. In our lab, we established a cell-based assay model to evaluate promising drugs for their ability to enhance the blood-brain barrier (BBB) integrity and improve clearance of amyloid-β (Aβ). Etodolac and α-tocopherol were selected as promising drugs for in vivo evaluation. Both drugs were tested separately and in combination for targeting multiple pathways including BBB function, neuroinflammation and oxidative stress in 5XFAD mice. Mice were divided into 4 group and treated with vehicle, etodolac, α-tocopherol, and combination of both drugs. Compared to either drug alone, the combination significantly enhanced the BBB function, decreased total Aβ load correlated with increased expression of major transport proteins, promoted APP processing towards the neuroprotective and non-amyloidogenic pathway, induced synaptic markers expression, and significantly reduced neuroinflammation and oxidative stress. In conclusion, this study highlights the significance of combination therapy to simultaneously target multiple disease pathways, and suggest the repurposing and combination of etodolac and α-tocopherol as a novel therapeutic strategy against AD.
Blood-Brain Barrier Disruption in Aged and Cerebral Amyloid Angiopathy Mouse Models	Euitaek. Yang, Amal Khalil. Kaddoumi	Auburn University, Harrison School of Pharmacy, Drug Discovery and Development, Pharmaceutics	Alzheimer’s disease (AD) is a neurodegenerative disorder with progressive hippocampal and cortical neuronal degeneration with dementia. Hippocampus and cortex are responsible for long- and short-term memorization, and thus neuronal degeneration in these regions could cause cognitive impairment. Aging is one of the risk factors to trigger blood-brain barrier (BBB) dysfunction. BBB plays a critical role in the clearance of brain waste products including Amyloid-β (Aβ); BBB disruption could alter these products clearance and allow entry of neurotoxic molecules to the brain at the same time. Reduced clearance of Aβ could result in its accumulation in the brain, which could trigger hippocampal and cortical neuronal degeneration. The purpose of this project is to compare the BBB integrity in young wild type, and 10 months old wild-type and CAA (cerebral amyloid angiopathy) mice. For this, we used immunostaining to monitor and compare BBB leakage by IgG extravasation assay. Our findings demonstrated the BBB leakage was highest in CAA mice brains compared to wild-type mice.
Novel drug delivery design and evaluation of Donepezil loaded 3D printed implant for improving compliance in Alzheimer’s patients	Manjusha Annaji, Sindhu Ramesh, Ishwor Poudel, T. Ramapuram, Muralikrishnan Dhanasekaran, Jayachandra R. Babu	Auburn University, Department of Drug Discovery and Development	Alzheimer’s disease (AD) is the most common form of dementia and is estimated to be affecting 5.7 million Americans and 44 million individuals globally. Therefore, World Health Organization recognized AD as a global health priority. Donepezil (cholinesterase inhibitor, cholinomimetic), which is available in the form of 5-10 mg tablet, is the most widely used drug for the treatment of mild to moderate AD currently. Rapid oral absorption of donepezil results in fluctuations of drug levels which causes several adverse drug reactions-ADR (nausea, vomiting, diarrhea, sleep disturbances, bradycardia and aggressiveness). Additionally, AD patients are vague and fail to comply with medications on a daily basis. Such non-compliance represents a widespread issue and often underestimated problem, which places a tremendous burden on the current and future healthcare system. The 3D printed implants are precise, therefore provide a superior mode of drug delivery with improved pharmacokinetic and pharmacodynamic parameters which results in enhanced quality of life of AD patients. Such implantable devices to the best of our knowledge are not available for AD therapy. Our present aim is to design a drug-eluting 3D printed implant at monthly intervals to provide steady state drug levels with minimal ADR. Our goal is to design and fabricate donepezil loaded novel biodegradable 3D printed implants. The implants will be characterized for its uniformity of drug content and thickness. In vitro release of the characterized implants will be performed to analyze the drug release kinetics and degradation mechanisms. Finally, in vivo evaluation of the donepezil loaded 3D printed implants will be conducted to determine the pharmacokinetic and pharmacodynamic parameters and the neuroprotective effectiveness in a valid animal model of AD. Keywords: Alzheimer’s disease, Donepezil, adverse drug reactions, non-compliance, 3D printed implant Acknowledgements: We acknowledge the support of funding from Presidential Award for Interdisciplinary Research (PAIR) grant.
Hispolon as a potential compound for the treatment of neurodegenerative disorders: A novel drug delivery approach	Ishwor Poudel, Manjusha Annaji, Sindhu Ramesh, T Ramapuram, Muralikrishnan Dhanasekaran, Jayachandra R Babu	Department of Drug Discovery and Development, Auburn University AL 36849	Hispolon, a polyphenolic compound isolated from Phellinus linteus has shown promising anti-inflammatory, antioxidant and antitumor activities. Hispolon is a structural analogue of curcumin, a pleiotropic polyphenolic derivate with tremendous potential for neuroprotective efficacy. The poor oral absorption confines the possibility of effective drug delivery and bioavailability. The incorporation of hispolon in clinical regimen for patients with neurodegenerative diseases like Alzheimer’s and Parkinson’s disease is a challenge, as patients tend to skip medications and fail to comply with the dosing schedule. Such non-compliance is often underestimated and results in a tremendous burden to healthcare system. Moreover, the relationship between hispolon and neuroprotective potential is not investigated till date. Use of implantable formulations is one of the best choices to avoid the above limitation. Implants when 3D printed could be personalized for precise drug delivery to patients with desirable therapeutic efficacy resulting in augmented drug bioavailability and reduced dosing frequency. Our goal is to fabricate drug eluting 3D printed implants for sustained delivery of hispolon for at least 4-6 weeks using thermoplastic extrusion technology. The physical characterization will involve drug loading, content uniformity and morphological evaluation. In vitro release of the characterized implants will be performed to exhibit the drug release kinetics and degradation mechanisms. The in vivo evaluation of the hispolon loaded 3D printed implants will be further conducted to elucidate the pharmacokinetic and pharmacodynamic parameters and the neuroprotective effectiveness in a valid animal model of Alzheimer’s disease. Acknowledgements: We acknowledge the support of funding from Presidential Award for Interdisciplinary Research (PAIR) grant.
Lysophosphatidic acid (LPA)-induces tau hyperphosphorylation through p38 MAPK	Sindhu Ramesh, Muralikrishnan Dhanasekaran, Timothy Moore	Auburn University, Drug Discovery and Development, Center for Neuroscience Initiative, Harrison School of Pharmacy	The number of Alzheimer’s disease (AD) cases – currently estimated to be greater than 5.4 million – is predicted to rapidly increase in the coming decades. Current AD treatments provide, at best, only modest and temporary symptomatic relief failing to alter the underlying pathophysiology, which leads to the progression of the disease. Lysophosphatidic acid (LPA), a bioactive phospholipid involved in inflammation, has been associated with development of AD. Hence, the aim of this study was to investigate whether LPA might promote AD pathology through tau hyperphosphorylation and identify the putative molecular signaling mechanisms by which LPA elicits this effect. Furthermore, this study tested whether atorvastatin can attenuate LPA-induced tau hyperphosphorylation. Using human SH-SY5Y cell lines differentiated with 10µm retinoic acid to achieve a neuronal phenotype, dose-dependent effects of LPA and LPA (+) atorvastatin on cell viability and dendritic morphology were determined. PrestoBlue™ cell viability assays showed LPA decreased cell viability at 5 µm and 10µm that was prevented with 1µm atorvastatin. LPA also caused neurite retraction in a time-dependent manner. Neurite retraction was accompanied by the phosphorylation of tau (Ser404 and Thr231. These effects were reversed by the p38 MAPK inhibitor SB203580, the JNK inhibitor JNK(i) II but not by the MEK (MAPK/ERK kinase) inhibitor, PD98059. The downstream effects including NF-kappaB transcription and AP-1 transcription will be determined. Taken together, these studies provide evidence for LPA as a tau-dependent inducer of neuronal degeneration involving the p38MAPK pathway. Prevention of the LPA-induced effects are accomplished by atorvastatin, and therefore these findings open a new avenue for exploring statins as potential therapeutics for AD
Investigation of ILK signalling deficits in prenatal alcohol rats exposed to 7,8-DHF	Claire E. Mullins, Dwipayan Bhattacharya, Sindu Ramesh, Manoj Govindarajulu, Jenna Bloemer, Subhrajit Bhattacharya, Manal Buabeid, Martha Escobar, Timothy Moore, Vishnu Suppiramaniam, Muralikrishnan Dhanasekaran	Auburn University, Department of Drug Development and Discovery	Through observation of rats in a fetal alcohol spectrum disorder (FASD) model, moderate prenatal alcohol exposure displayed significant deficits in both learning and memory tasks, in addition to reduced synaptic plasticity. Our previous studies have shown these plasticity deficits are associated with reduced ILK activity and increased GluR2 AMPA receptors at the synapse. We hypothesized that enhancing the ILK activity might ameliorate both the behavioural and plasticity deficits that were recognized in the FASD model. 7,8-dihydroxyflavone (7,8-DHF) acts as a potent agonist for Brain Derived Neurotropic Factor (BDNF) receptor Tyrosine receptor kinase B (TrkB). Therefore, we hypothesize that intraperitoneal administration of 7,8-DHF in FASD rats, during the early postnatal days, may restore the alcohol associated memory deficits through increasing ILK activity and improve synaptic plasticity. Consequently, we investigated the neuroprotective effects that 7,8-DHF in the FASD model. 7,8-DHF led to a significant positive impact upon the behavioural deficits associated with FASD by causing a significant increase in the ILK activity.
A Magnetic Nanoplatform for Identification and Isolation of Compounds Targeting Ion Channels	Jesse Horne (1), Shomit Mansur (1), Dr. Lukasz Ciesla (2), and Dr. Yuping Bao(1*)	(1) Department of Chemical and Biological Engineering, The University of Alabama, Tuscaloosa, AL 35401, USA (2) Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL 35401, USA *Indicates the corresponding author and can be reached at ybao@eng.ua.edu	Natural products, such as plant extracts, are rich in many bioactive compounds, but a significant challenge is the screening of complex mixtures. Traditional screening methods often require isolation of individual compounds from natural resources before analysis. Here, cell membrane-encapsulated iron oxide superparticles (a cluster of nanoparticles)-CMSPs were developed for the identification and isolation of molecules targeting ion channels, such as sodium ion channels Nav. Ion channels are often used as targets for pain-killing drugs, but many drug screening techniques are unable to select for specific ion channel subtypes. MDA-MB-231 cells and the brain metastatic cell line MDA-MB-231 BR were chosen as a model system to validate this nanoplatform because these cell lines are known to overexpress Nav 1.5 sodium ion channels. The CMSPs were characterized through scanning electron microscopy, transmission electron microscopy, and dynamic light scattering to observe a morphology change with the self-assembly of the cell membranes around the superparticles. The effects of four superparticle coatings (polyethylene glycol, pectin, polyacrylic acid, and polyvinylpyrrolidone) on the ion channel state evaluated to systematically investigate the surface chemistry effects on ion channel states (open, close, and inactive). The control of ion channel states will allow for the selective identification of molecules targeting different regions of the ion channels, such as voltage-sensing or pore-forming domains. This work will significantly advance our fundamental understanding of the ion channel immobilization and their use as targets for pain dug lead identification, with the ultimate goal of contributing to the development of novel pain management pharmaceuticals.
The long non-coding RNA Neat1 in epigenetic regulation of astrocytes in an Alzheimer’s disease model	Ashleigh B. Irwin1,2, Farah D. Lubin2	1NIH Medical Scientist Training Program, University of Alabama at Birmingham, Birmingham, Alabama, USA 2Department of Neurobiology, University of Alabama at Birmingham, Birmingham, Alabama, USA	Loss of normal epigenetic regulation in the brain is strongly associated with neuropsychiatric conditions, and may provide a framework for understanding the complex factors that contribute to memory dysfunction in Alzheimer’s disease. We recently demonstrated that the long non-coding RNA Neat1 mediates histone methylation and age-related memory impairments, and that reducing Neat1 expression rescues these memory deficits. These studies prompted us to ask if Neat1 contributes to AD-associated cognitive decline and if so, if Neat1 might serve as a therapeutic target. Previous studies have shown that Neat1 expression is significantly elevated in hippocampal astrocytes of patients with Alzheimer’s disease. Using magnetically activated cell sorting, we have been able to isolate microglia and astrocyte fractions from an aged mouse model of AD. Preliminary data suggests elevated Neat1 expression in the hippocampus of the hAPP-J20 mouse model of AD, both in whole extracts and in cell-type specific analyses, similar to that seen in the human condition. Using this model, we are testing the hypothesis that Neat1 epigenetically regulates astrocyte-specific genes involved in hippocampus-dependent memory formation.
Intravenous AAV gene therapy improves lifespan and clinical metrics in feline Sandhoff Disease	Anne S Maguire, Elise B Diffie, Amanda L Gross, Hannah G Lahey, Miguel Sena-Esteves, Douglas R Martin	Scott-Ritchey Research Center, Department of Anatomy, Physiology, & Pharmacology, College of Veterinary Medicine, Center for Neuroscience Initiative, Auburn University; Department of Neurology, Horae Gene Therapy Center, University of Massachusetts Medical School	Sandhoff Disease (SD) is a neurodegenerative lysosomal storage disease that causes the death of severely affected children before 5 years of age. This autosomal recessive disease results in the dysfunction of the enzyme ß-hexosaminidase (Hex) and the subsequent accumulation of GM2 ganglioside in neuronal lysosomes. In previous studies in a feline model of infantile SD, intracranial administration of gene therapy in the form of an adeno-associated viral (AAV) vector quadrupled lifespan and increased quality of life. In an attempt to reduce the risk of intracranial surgery and achieve greater systemic vector distribution, we treated 11 SD cats intravenously (IV) at one month of age with a bicistronic AAV9 vector expressing Hex. These cats were divided into three groups: low dose (5e13/kg) short-term (n=3), low dose long-term (n=4), and high dose (2e14/kg) long-term (n=4). Animals in the short-term group were euthanized 16 weeks post-treatment, while cats in the long-term group were followed to a predetermined humane endpoint (inability to stand). While untreated SD cats live to 4.3±0.2 months, cats treated with the low and high doses lived to 8.3±1.2 and 11.1±1.5 months, respectively, with one cat in the high dose group currently alive at 15.6 months. In-life assessments revealed clear clinical benefit of AAV treatment, with the most dramatic improvement seen in the reduction of tremors, the most debilitating feature of feline SD. Bimonthly neurological examinations demonstrated markedly improved quality of life in AAV-treated animals. Cerebrospinal fluid (CSF) levels of aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were decreased both at 16 weeks post-treatment and at long-term endpoints, indicating a reduction of cell damage within the central nervous system. These results support the efficacy of IV delivery of a bicistronic AAV vector, especially at a high dose, for improving lifespan and quality of life in a feline model of SD.
Phosphoprotein Enriched in Astrocytes-15 kD (PEA15) affects cellular glucose uptake and energetics in the brain of domestic cats	Towns, T.J., Watanabe R., Brinker, E.J., Bashir, A., Maguire A.S., Gross, A.L., Martin, D.R., Graff, E.C.	Auburn University College of Veterinary Medicine, Scott-Ritchey Research Center	Abnormalities in brain glucose metabolism underlie many neurological diseases. Recently, our lab discovered that a loss-of-function mutation in Phosphoprotein Enriched in Astrocytes-15kD (PEA15), an intracellular adaptor protein, has an essential role in gyrencephalic neurodevelopment. The absence of PEA15 results in severe microcephaly and polymicrogyria in domestic cats. Several studies suggest that differential PEA15 expression alters glucose uptake and metabolism; however, previous work focused on peripheral, insulin-dependent tissues. The role of PEA15 in glucose uptake and metabolism has not been characterized in the central nervous system. Therefore, the objective of this study was to investigate the effects of a PEA15 loss-of-function mutation on neurodevelopment and cellular energetics in a gyrencephalic animal. Developmental milestones, clinical assessments, and metabolic parameters in the serum and cerebrospinal fluid (CSF) were collected over a 2-year period on PEA15-/- and PEA15+/+ cats. At 1 year of age, 31P-MRS of the brain was performed to evaluate in vivo creatine kinase flux (CK-flux) as an indirect measure of cell energetics. Additionally, a fluorometric glucose uptake assay was performed on primary cultured fibroblasts from affected (PEA15-/-) and unaffected (PEA15+/+) cats. No metabolic differences were noted between groups in the CSF or serum. The cerebral CK-flux is decreased by ~60% in affected cats compared to unaffected at 1-year of age. Cultured PEA15-/- fibroblasts had significantly decreased basal glucose uptake at 6 hours (P < 0.001) and 12 hours (P < 0.001) compared with fibroblasts from unaffected cats. These findings suggest that there is altered cellular glucose metabolism and cerebral cellular energetics associated with PEA15 loss-of-function. However, these changes are likely compensated systemically by alternate metabolic pathways. Future studies will determine if changes in PEA15 neurometabolism contribute to the neurodevelopmental deficits observed in PEA15-/- cats.
Role of Prenatal Synthetic Cannabinoid Exposure on the Cerebellum of Adolescent Rat Offspring	Justin Hall1, Priyanka D. Pinky2*, Mohammed Majrashi3*, Fujihashi Ayaka2, Jenna Bloemer2, Vishnu Suppiramaniam1#, Muralikrishnan Dhanasekaran,1#	1College of Science and Mathematics, Auburn University, Auburn, Alabama, USA 2Department of Drug Discovery and Development, Auburn University, Auburn, Alabama, USA 3Department of Pharmacology, Faculty of Medicine, University of Jeddah, Jeddah, 23881, Saudi Arabia.	Cannabinoid is an illicit recreational drug used by some women during pregnancy to attenuate morning sickness. However, prenatal cannabinoid exposure produces enduring cognitive, locomotor, and behavioral deficits in the offspring by affecting neural circuitry and various signaling mechanisms that are responsible for learning and memory. The effect of prenatal cannabinoid exposure on cerebellum are still not well elucidated. Cerebellum plays an important role in balance, motor control, as well as some cognitive processes such as attention, language, emotional functions and procedural memories. Therefore, the current study evaluated the effects of prenatal cannabinoid (synthetic cannabinoid agonist – WIN55, 212-2) exposure on the cerebellar markers of oxidative stress (reactive oxygen species-ROS, Nitrite-NO, lipid peroxide), mitochondrial function (Complex-I and IV activities), apoptosis (Caspase-1, Caspase-3, ERK, p-JNK, P38), and the cerebellar signaling molecules associated with cannabinoid (CB1 receptor), glutamatergic (GluA1, GluN2A receptor), and dopaminergic neurotransmission (monoamine oxidase (MAO) & tyrosine hydroxylase activities), and synaptic plasticity (AKT,GSK3β, ILK). Administration of WIN55, 212-2 during pregnancy altered the markers of oxidative stress by significantly reducing nitrite content, without any alteration in ROS generation and lipid peroxide content. An anti-apoptotic effect has been observed by increased expression of P38 expression. On the other hand, pro-apoptotic factor caspase-3 activity, pERK, and pJNK expression were significantly decreased. CB1 and GluA1 receptor expression remained unchanged; however, the GluN2A receptor expression was significantly reduced. With regard to the dopaminergic neurotransmission, a significant decrease in the MAO activity with no change in tyrosine hydroxylase activity has been observed. Altogether, data from this study provides evidence for cerebellar alterations following prenatal synthetic cannabinoid exposure.
Regular Cannabis Use is Associated with Poorer Pain Outcomes Inside and Outside the Laboratory: Results from Experimental and Naturalistic Assessments	Julio A. Yanes, Meredith A. Reid, Lauren Y. Atlas, Jared W. Younger, Raul Gonzalez, and Jennifer L. Robinson	Auburn University Center for Neuroscience Initiative Auburn University MRI Research Center National Center for Complimentary and Integrative Health University of Alabama Birmingham Florida International University	Substantial efforts have been directed toward understanding the complex relationship between cannabis and pain, yet we have many (often conflicting) theories regarding cannabinoid analgesia. Critically, although cannabis’s acute effects on pain have received considerable attention, long-term effects have been severely understudied. Here, we leveraged experimental and naturalistic approaches to provide enhanced understanding about regular cannabis use and pain. Two, inter-related studies were conducted: a laboratory study and an internet study. During the laboratory study, we measured acute pain in recreational cannabis users and cannabis non-users to test two hypotheses: (i) that subjective pain ratings are lower among users and (ii) that maximum pain tolerance is greater among users. During the internet study, we retested observed associations using more ecologically-relevant endpoints in a separate sample. When taken together, our results support the following conclusions: (i) recreational cannabis is associated with lower maximum pain tolerance and (ii) users report greater pain-related impairment outside the laboratory. Despite meta-analytic evidence that cannabis treatments reduce experimental pain and clinical pain in individuals without cannabis use histories, our findings are consistent with recent reports documenting poorer post-operative pain outcomes among chronic users. Given recent advancements in medical, political, and societal attitudes toward cannabis, it has become increasingly important to consider potential motivations (e.g., self-medication) or unintended consequences (e.g., hyperalgesia) associated with regular, long-term use.
Measurement of Blood Brain Barrier Permeability in Human Brain using Magnetization Transfer Effect at 7T.	Sultan Z. Mahmud, Thomas S. Denney, Ronald J. Beyers, Adil Bashir.	Auburn University, Department of Electrical and Computer Engineering. Auburn University, MRI Research Center.	Blood brain barrier (BBB) plays a very important role in regulating water and nutrients delivery between vascular circulation and central nervous system (CNS). Any disruption in the blood brain barrier may cause the alteration of normal functional activity of the nervous system. Many CNS diseases such as Multiple Sclerosis, Alzheimer’s disease etc. are associated with compromised BBB. The techniques currently available to measure BBB permeability are prone to certain limitations and potential side effects. In this study we demonstrated a non-invasive technique of evaluating BBB permeability at 7T using the magnetization transfer (MT) effect on endogenous water labeled by arterial spin labeling (ASL) technique as a perfusion tracer. Perfusion measured with and without MT can be compared to estimate water extraction fraction and BBB permeability. FAIR QUIPSS II ASL technique was used for quantification of perfusion with and without MT. 6 healthy subjects partcipated the study. Average perfusion in the gray matter and white matter was 66 ± 11 mL/100g/min and 41 ± 14 mL/100g/min respectively. Average extraction fraction in white and gray matter was 0.957 ± 0.02 and 0.916 ± 0.03 respectively. Average permeability surface area product in white and gray matter was 129 ± 43 mL/100g/min and 163.5 ± 53 mL/100g/min respectively.
Trifluoromethylphenyl (TFMPP) derivatives affects dopaminergic, hippocampal and PS70 cells	Zhao, Mingliu; Harshan, Aisha; Alturki, Mansour; Almaghrabi, Mohammed; Ramesh, Sindhu; Govindarajulu, Manoj; Alghenaim, Fada; Smith, Forrest; Deruiter, Jack; Clark, Randall; Dhanasekaran, Muralikrishnan	Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University	Background: In the United States, 1 out of every 20 deaths is associated to substances of abuse. Furthermore, drug abuse has become a huge global problem. Trifluoromethylphenyl (TFMPP) derivatives (2, 3, 4) belong to class of Piperazine designer drug and it is abused as an alternate to the illicit drug-MDMA. Designer drugs have shown to induce cognitive impairment and dopaminergic neurotoxicity. Methods: MTT cell viability assay was used to assess the effect of 2-TFMPP, 3-TFMPP and 4-TFMPP on dopaminergic, hippocampal and PS70 cells. Results: 2-TFMPP, 3-TFMPP and 4-TFMPP dose-dependently and time-dependently decreased the cell viability significantly of the dopaminergic, hippocampal and PS70 cells Future Studies: Our future study is to evaluate the effect of these designer drugs on movement and memory disorders.
“Piperazine” Designer Drugs affect the cells producing Amyloid-beta	A.Harsha, , M. Zhao, M. Alturki, M.Almaghrabi, S.Ramesh, M. Govindarajulu, F.Alghenaim, F. Smith, J.Deruiter, C Randall, M.Dhanasekaran	Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University	Designer drugs are illegally synthesized to produce the psychostimulatory effects similar to the banned psychotic drugs of abuse. Piperazine designer drugs were formulated due to the legal banning of Amphetamine (Scheduled Substance of Abuse). Apart from their psychostimulatory effects, designer drugs induce mental, movement, and memory-related adverse effects. The current global peril is the distressing rise in the number of Alzheimer’s disease (AD) patients. The extracellular senile plaques that accumulate in the brain of Alzheimer’s patients are caused by Amyloid-beta (Aβ) through sequential cleavages of amyloid-β precursor protein (APP) by β and γ-secretase. Interestingly, drugs that modulate Aβ pathway are considered to be one of the most promising avenues for treating AD. However, the role of Piperazine on the pathophysiology of Aβ metabolism is not well elucidated. The purpose of this study is to investigate the effects of Piperazine derivatives on Aβ metabolism. PS70 cells-Chinese Hamster Ovarian (CHO) cells (carrying the amyloid precursor protein-APP and Presenilin PS1 mutation) are valid in vitro model for this study. Piperazine derivatives dose-dependently inhibited the PS70 cell proliferation. Furthermore, based on our drug-based computational analysis, we will elucidate the interaction of Piperazine derivatives with various markers of (Aβ) metabolism. Thus, our current study will help to shed more light on the effects of Piperazine derivatives on the Aβ pathology in AD.
“Kainic acid”- A potent neuroexcitatory amino acid agonist may not be a valid model to assess dopaminergic neurotoxicity.	Alghenaim, Fada; Alturki, Mansour; Almaghrabi, Mohammed; Harshan, Aisha; Zhao, Mingliu; Ramesh, Sindhu; Smith, Forest; Clark, Randall; Deruiter, Jack; Dhanasekaran, Muralikrishnan	Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University	Alteration of dopaminergic neurotransmission (synthesis, storage, release, degradation, reuptake, and receptor interaction) is associated with Parkinson disease. Numerous neurotoxins such as 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), 6-hydroxydopamine, divalent metals (iron), insecticide / pesticides (salsolinol, diquat) have known to significantly induce dopaminergic neurotoxicity. Kainic acid is an epileptic agent that induces seizures in animals through binding with kainate receptors. In this study, we compared the dopaminergic neurotoxicity of kainic acid with the active metabolite of MPTP using N27 dopaminergic neuronal cells. Kainic acid exhibited significantly lower dopaminergic neurotoxicity as compared to MPP+ (1-methyl-4-phenylpyridinium).
Can computational modelling predict the neurotoxicity of designer drugs?	Ramapuram, Hariteja; Alturki, Mansour; Almaghrabi, Mohammed; Majrashi, Mohammed; Fujihashi, Ayaka; Ramesh, Sindhu; Govindarajulu, Manoj; Moore, Timothy; Forrest, Smith; Clark, Randall; Deruiter, Jack; Dhanasekaran, Muralikrishnan	Department of Drug Development and Discovery, Harrison School of Pharmacy, Auburn University, Auburn, AL	Computational modeling has shown to predict the pharmacological activity and the neurotoxicity of various drugs. Designer drugs with low toxicity can be a potent therapeutic tool to treat various peripheral impairments, endocrine disorders, and neurological diseases. Hence, in this study, we used new and novel substances of abuse and compared their computational modeling effects (Qikpro) with their in vitro effect on cell viability. 3-Trifluoromethylphenylpiperazine (TFMPP) derivatives (2-TFMPP, 3-TFMPP, 4-TFMPP, 2-TFMBzPP, 3-TFMBzPP, 4-TFMBzPP and BZP) were used in the present study. Based on Lipinski’s rule of five, Jorgensen’s rule of three, QPlogBB and the QPPCaco, the TFMPP derivatives can undergo passive diffusion and can be highly absorbed after oral administration (can be bioactive orally). PISA and FOSA values can be used to correlate with the neurotoxicity of a drug/chemical. TFMPP derivatives have lower cLogp, HBD and HBA values. Therefore, these designer drugs can cross the BBB and possibly exhibit significant neuropharmacological effects in the central nervous system. TFMPP derivatives exhibited significantly higher neurotoxicity as compared to the TFMBzPP derivatives.
Chemotherapeutics can increase the risk of dopaminergic neurotoxicity	Anna Solomonik, Darshini Desai, Mohammed Majrashi, Mohammed Almaghrabi, Kodye Abbott, Sindhu Ramesh, Manoj Govindarajulu, Satyanarayana Pondugulu, Timothy Moore, Muralikrishnan Dhanasekaran	Department of Drug Development and Discovery, Harrison School of Pharmacy	Chemotherapy-induced cognitive impairment, also known as “chemo brain”, is a medical complication of cancer treatment that is characterized by a general decline in cognition affecting visual and verbal memory, attention, complex problem-solving skills, and motor function. It is estimated that one-third of patients who undergo chemotherapy treatment will experience cognitive impairment. Alterations in the release and uptake of dopamine and serotonin neurotransmitters that play important roles in cognition could potentially contribute to impaired intellectual performance in those impacted by chemo brain. Chemotherapeutics can increase oxidative stress, decrease mitochondrial function, and increase apoptosis which causes dopaminergic neurodegeneration leading to motor disorders such as Parkinson’s disease. This study was done to investigate in vitro dopaminergic neurotoxic effects of the chemotherapeutics Doxorubicin and cyclophosphamide. Rat dopaminergic neuronal cells (N27) were used for in vitro studies to assess the neurotoxicity. Chemotherapeutics Cyclophosphamide and doxorubicin exhibited dose-dependent and time-dependent in vitro dopaminergic neurotoxicity. Chemotherapeutics doxorubicin and cyclophosphamide induced oxidative stress and apoptosis (increased BAX expression) in the dopaminergic neurons without affecting the mitochondrial functions. With regards to oxidative stress, cyclophosphamide and doxorubicin increased the generation of ROS and nitrite content resulting in lipid peroxidation. Cyclophosphamide and doxorubicin depleted glutathione, decreased catalase activity, and increased glutathione peroxidase, SOD, and MAO activity. Future studies to assess the in vivo neurotoxic effects of cyclophosphamide and doxorubicin using a rodent animal model will be performed. Therefore, this study supports the need for additional neurochemical, behavioural, and biochemical analysis to identify the underlying mechanisms of chemotherapy-induced cognitive disorders.
Scutellaria lateriflora: An innovative botanical to treat inflammation driven neurodegenerative diseases	Kelsey Schoenmeyer, Madhukar Lohani, Mrigendra Rajpoot, Sindhu Ramesh, Manoj Govindarajulu, Mohammed Majrashi, Eddie Fahory, Timothy Moore, Dennis Shannon, Frederik van Ginkel, Dean Schwartz, Barbara Kemppainen, Muralikrishnan Dhanasekaran	Auburn University Harrison School of Pharmacy Department of Drug Discovery and Development, Auburn University College of Veterinary Medicine Department of Anatomy Physiology and Pharmacology, Auburn University College of Veterinary Medicine Department of Pathobiology, Auburn University College of Agriculture Department of Agronomy and Soils, South Dakota State University Department of Veterinary and Biomedical Sciences	Scutellaria lateriflora is a native plant of North America that contains a larger amount of chemically active flavonoids than other commonly used species of the plant genus Scutellaria. To date, very limited research studies have been done to measure neuroprotective activities of S. lateriflora. The purpose of this study was to determine the neuroprotective mechanisms (immunomodulatory and anti-inflammatory) of S. lateriflora. Experiments were designed to measure the effects of an alcoholic extract of S. lateriflora (SLE) on pro-inflammatory enzyme (cyclooxygenase-1-COX-1, cyclooxygenase-2-COX-2) and 15 Lipoxygenase (15-LOX) activities, activation of T lymphocytes and major histocompatibility complex class-II (MHC-II) expression in bovine monocyte-derived dendritic cells (MoDCs). The SLE inhibited COX-1, COX-2 and 15 LOX activities but did not affect T cell proliferation or MHC-II expression on MoDCs or pro-inflammatory cytokines production by T cells. Our results established the anti-inflammatory mechanisms of SLE, which validates the potential possibility of treatment for various inflammation driven neurodegenerative diseases.
Are “Designer drugs” a global liability and peril?	Katherine Bricco, Sindhu Ramesh, Manoj Govindarajulu, Jack Deruiter, Randall Clark, Muralikrishnan Dhanasekaran	Auburn University, Harrison School of Pharmacy, Department of Drug Development and Discovery	Background: Designer (or synthetic) drugs are defined as “illicitly produced with the intent of developing substances that differ slightly from controlled substances in their chemical structure while retaining their pharmacological effects”. Designer drug production initially arose as legal regulatory bodies began to ban and regulate various chemical compounds due to their various psychostimulatory effects. Currently many drug manufacturers produce drugs of abuse in clandestine labs and distribute them illicitly to mimic and/or enhance psychostimulatory effects. As of 2017, United Nations determine that 5.5% of the global population aged 15-64 had used illicit drugs. Over the past years the use of illicit drugs globally has steadily increased. As the use of illicit drugs increases so have the increase in development and use of designer drugs. With the rise of synthetic or designer drug abuse and substantial diversity in adverse effects among formulations available there is an increase in crime and health concern on global populations. Methods: Combined meta-analysis of statistics in illicit drug use and abuse trends. The primary conclusion was determined by analysis of lifetime and first-time use of illicit drug trends from global safety and anti-drug databases. Secondary conclusions and reasoning for future research were determined by overdose trends compiled by the CDC. Results: The most common drugs of abuse in the past 5 years included: cocaine, methamphetamine, marijuana, heroin, and hallucinogens. Of these common illicit drugs there are over 200 identified synthetic or designer versions of the drug circling the globe. Over the past years as the increase in designer drug production increases there has also been a large jump in the number of overdose deaths due to designer drugs. This increase in the number of overdose death leads to an increase in global concern and need for further research regarding adverse health outcomes of these unregulated substances. Conclusion: Of the most common illicit drugs of abuse in the past 5 years, all have some form of synthetic versions available in the market which are being abused. With the upsurge in synthetic drugs available there is an increase concern regarding adverse health outcomes and how to adapt legal regulation.
LEC-specific P301L tau results in synaptic alterations and learning and memory deficits	Setti, Sharay, E.; Bloemer, Jenna, E.; Pinky, Priyanka, D.; Pfitzer, Jeremiah, C.; Dogan, Marrisa, H.; Suppiramaniam, Vishnu; Reed, Miranda, N.	Auburn University, Center for Neuroscience Initiative, Harrison School of Pharmacy	Alzheimer’s Disease (AD) is a fatal neurodegenerative disease that disrupts and destroys normal neuronal function via the formation of beta-amyloid plaques and neurofibrillary tangles made of hyperphosphorylated tau protein. One of the first brain regions to exhibit tau pathology is the lateral entorhinal cortex (LEC), with pathology “spreading” or “propagating” to connected regions, such as the hippocampus, as the disease progresses. To better understand the mechanisms underlying tau propagation, in experiment 1 we first injected an adeno-associated viral (AAV) vector encoding human tau with the P301L mutation into the LEC of healthy mice. After four weeks of LEC-P301L tau expression, we assessed for learning and memory impairments using memory tasks (object recognition and trace fear conditioning) designed to be LEC-dependent. At the conclusion of memory testing, we assessed for the presence of tau pathology using immunohistochemical staining of brain slices. Our results indicate that compared to controls, LEC-P301L mice exhibit subtle deficits in learning and memory that is associated with tau pathology in the LEC. LEC-P301L also mice exhibit impaired synaptic plasticity as indicated by a reduction in long-term potentiation, a cellular correlate of learning and memory. Because evidence from the literature suggests that neuronal activity may mediate release of pathological tau from neurons, thereby underlying the spread of tau pathology from the LEC to the hippocampus, in experiment 2 we delivered bilateral injections of AAV vector encoding human tau with the P301L mutant to the LEC. Then, we used optogenetics to stimulate neuronal activity in one hemisphere. Tau pathology spread further from the site of injection in the stimulated versus non-stimulated hemisphere, as indicated by presence of tau pathology within the hippocampus. Preventing tau spread may ultimately prove to be a promising therapeutic target for treatment of Alzheimer’s disease.
The thyroid effect on brain transcriptome	Huifei Sophia Zheng , Chen-Che Jeff Huang	Department of Anatomy, Physiology & Pharmacology, College of Veterinary Medicine, Auburn University	Thyroid hormones are important in brain development by regulating cell migration and differentiation, synaptogenesis, and myelination. Clinical hypothyroidism and hyperthyroidism have long been recognized as potentially reversible causes of cognitive impairment. Thyroid dysfunction has emerged as a possible risk factor for the development of irreversible dementia, with several epidemiological studies implicating both hypo- and hyperthyroidism. Thyroid hormone can also stimulate the expression of the selective Alzheimer’s disease indicator-1 (Saladin-1) gene, which has been shown to have lower abundance in the areas affected by Alzheimer’s disease. To understand how thyroid hormone affects the brain cortex at the transcriptome level in adult mice, we used RNAseq to analyze B6 male mice under four different thyroid conditions including (1) short-term T3 treatment group and (2) the saline control group; (3) long-term hypothyroidism (hypo-T) and (4) long-term hyperthyroidism (hyper-T). Our data showed that most T3-responsive genes are low-expressing genes. Only 1105 T3-responsive genes had the expression level above one RPKM. Eighty-four (84) of them showed a >2-fold change. Also, the expression of Alzheimer’s diseases-associated genes (430 genes) were relatively stable among different thyroid conditions. In the field of studying brain development and the thyroid hormone action, it is suggested that the gene regulation by thyroid hormone follows a strict temporal and regional specificity; most genes are sensitive only during a limited time period of postnatal development, and some genes are under thyroid hormone control only in certain brain regions. Our data provide a transcriptome-level evidence on the current understanding of the thyroid hormone action in the adult brain.
Gut microbiota metabolite, TMAO, induces microglial activation and senescence in rodent models with memory impairment.	Ian Steinke, Manoj Govindarajulu, Robert Rella, Priyanka Das Pinky, Nila Ghanei, Taylor Schaedig, Melissa Boersma, Forrest Smith, Vishnu Suppiramaniam and Rajesh Amin	Dept of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University, Auburn AL. Director of Chemical Laboratory, Chemistry & Biochemistry, Auburn University, Auburn AL.	Research has intensified in the area of the gut microbiome and how it is linked to various disease pathologies. Dysbiosis of these bacteria can occur when dietary intake influences an imbalance in healthy bacteria and has been observed in Diabetic and Alzheimer’s patients. Dysbiosis metabolites are further metabolized by the liver to produce Trimethylamine-N-oxide (TMAO). This novel biomarker for atherosclerosis and renal fibrosis is now being investigated for its association with the development of Alzheimer’s disease (AD). Neuroinflammation is a common factor associated with AD and Diabetes for the development of memory impairment and neuronal dysfunction. In the current study we will show that TMAO induces microglial activation leading to neuronal inflammation and cellular senescence. Quantification of TMAO levels from aged 3xTg-AD and diabetic (db/db) mice was accomplished by LC-MS analysis. Evidence of senescence was measured by beta gal staining in aged (8 month) 3XTg-AD mice and Diabetic mice. Microglial activation patterns by TMAO are measured by flow cytometry and western blot. Electrophysiological recordings for long term potentiation in mouse brain slices incubated with TMAO for 2 hours demonstrated altered plasticity and enhanced microglial activation. We have observed that TMAO is elevated in Diabetic and AD mice as disease progresses. TMAO also induces significant activation of microglial cells in vitro. Further, elevated levels of hepatic flavin-containing monooxidase 3 (FMO3), are observed in diabetic and 3xTg-AD mice. Our data is currently preliminary and further evaluation will determine the significance of TMAO to its related metabolic pathway in association with neuro-inflammation and cognitive impairment in Diabetes and AD. Second, FMO3 is central in TMAO metabolism and has been correlated to hepatic insulin resistance. This connection of TMAO with insulin sensitivity and glucose utilization is being investigated with novel FMO3 inhibitors.
SELECTIVE PPAR-γ AGONISM ATTENUATES NEURO-INFLAMMATION AND IMPROVES BRAIN ENERGY DYSREGULATION IN A MOUSE MODEL OF ALZHEIMER’S DISEASE.	Manoj Y Govindarajulu, Nila Ghanei, Sieun Yoo, Ian Steinke, Taylor Schaedig, VIshnu Suppiramaniam, Rajesh Amin	Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University Center for Neuroscience Initiative, Auburn University	Neuroinflammation and brain metabolic dysfunction are well-established feature of AD, that can be observed potentially decades prior to the development of AD symptoms. Thiazolidinediones (TZDs) are insulin sensitizing peroxisomal proliferator activating receptor-gamma (PPARγ) agonists and are promising agents for improving pathologies of AD. However, they display poor blood brain barrier (BBB) permeability, thus requiring high dosing and are associated with several adverse side effects. We have developed a novel PPARγ (AU-9) in-silico that avoids the unwanted side effects of current PPARγ agonist, including edema and effects on the heart. Furthermore, AU-9 displays enhanced BBB permeability, thus requiring lower concentrations to promote protection against AD. We hypothesize that our lead compound (AU-9), a selective dual Peroxisome proliferator-activated receptor γ/δ modulator improves neuroinflammation and energy dysregulation associated with AD in a mouse AD model. Six months old triple-transgenic AD (3xTg-AD) and C57BL/6J control mice were treated with AU-9 for one month. Hippocampal gene expression analysis for neuroinflammation and energy regulation was performed by Nanostring gene array. Hippocampal lysates were analyzed to determine the effect of AU-9 on various inflammatory and insulin signaling pathways; Immunohistochemistry to determine markers of inflammation and In vitro studies utilizing murine BV2 microglial cells to study the signaling mechanisms involved improving neuroinflammation and insulin signaling pathway. Gene expression analysis indicated decreased mRNA expression of various inflammatory genes and upregulation of genes involved in energy regulation with AU-9. Biochemical assays showed AU-9 attenuated p38 MAPK signaling, decreased microglial activation and improved PI3K-Akt pathway both in murine BV2 microglial cells and in 3xTgAD mice. Our results indicate that AU-9 offers potential therapeutic benefits for mitigating pathologies in AD.
Synaptic Vesicle Dynamics at Single Presynaptic Active Zones	Abhishek P Shrestha, Thirumalini Vaithianathan	University of Tennessee, Department of Pharmacology	Synapses of retinal bipolar cells and photoreceptor cells are characterized by special structures at the site of neurotransmitter release called synaptic ribbons. Although we know a great deal about ribbon synapses, it remains unclear how the ribbon supports distinct kinetics of neurotransmitter release. By exploiting the direct approach of tracking single vesicles at ribbons in living neurons via high-resolution optical imaging, we have resolved long-standing questions about the fate of vesicles on the ribbon prior to fusion, at fusion sites, and after fusion. To track single vesicles, we targeted the photoactivatable fluorescent protein to synaptic vesicles by fusing it to the C-terminus of the vesicle membrane protein vesicular glutamate transporter 1. To detect fusion of ribbon-associated vesicles, we generated transgenic zebrafish that express the exocytosis reporter Synaptophysin-pHluorin fusion protein. We used two-color laser scanning methods that allowed single labeled vesicles to be observed throughout the full extent of the ribbon in voltage-clamped synaptic terminals while the ribbon and cell border were imaged with a second fluorescent label. Our results suggest a dual role for the synaptic ribbon in neurotransmitter release, with the ribbon serving as both a conduit for diffusion of tethered synaptic vesicles and a platform for vesicles to fuse distal to the plasma membrane. Our approach also revealed precisely when and where a fusion event occurs and the rate of clearance of fused synaptic vesicles after fusion. In summary, the new approach that we introduced in this study provided new information about the nanoscale dynamics of single vesicles and allowed visualization of the events leading up to and following fusion itself. Our results set the stage for models of synaptic vesicle trafficking and fusion at the ribbon active zone that incorporates both calcium signaling and vesicle trafficking.
Single Channel Recordings of Synaptic AMPA Receptors from Epileptic Human Temporal Lobe	Warren D. Smith, Larry W. Ver Hoef, Jenna E. Bloemer, Vishnu Suppiramaniam	Auburn University, Department of Drug Discovery and Development, Center for Neuroscience Initiative University of Alabama at Birmingham, Department of Neurology	Using a novel technique to record synaptic ion channel activity, this project has produced the first single channel recordings of synaptic AMPA receptor activity from the brain of a living human patient. Samples were obtained at the University of Alabama at Birmingham (UAB) during a focal resection in a patient with refractory epilepsy. Synaptosomal fractions were biochemically isolated on-site, and recordings were performed at the Pharmacy Research Building at Auburn University. Receptors from the unafflicted cortex were compared to diseased tissue from the hippocampus foci in an attempt to discover possible functional characteristics of individual AMPA receptors contributing to this patient’s epilepsy.
Ayurvedic botanical exhibits Anti-Alzheimer’s effect by exhibiting significant neuroprotective effects	Mentlick, Kassandra; Ahuja, Manuj; Buabeid, Manal; Ramesh, Sindhu, Govindarajulu, Manoj; Almaghrabi, Mohammed; Alturki, Mansour; Smith, Forrest; Suppiramaniam, Vishnu; Moore, Timothy; Dhanasekaran, Muralikrishnan	Department of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University	Centella asiatica (CA) is a well-known medicinal herbal drug traditionally used in folklore Indian Ayurvedic medicinal system and in other parts of Asia for more than hundred years to treat a plethora of ailments. The whole aerial portion of the plant is highly rich in various biologically active compounds. Several lines of evidence report antioxidant and neuroprotective actions of Centella but the mechanism of action is unknown. Moreover, recent reports as well as ancient texts mentions cognition enhancement as one of the beneficial medical properties of Centella. In the current study we commercially obtained a concentrated extract of Centella asiatica containing 20% asiaticoside and used it to elucidate the neuroprotective and antioxidant action against well-known pro-oxidants and neurotoxin in two neuronal cell lines. In addition, we also investigated the molecular mechanisms related to neuroprotection as well as mnemonic activities in the hippocampal neuronal cell line H19-7. In addition to preliminary characterization of the current extract, the neuroprotective action of Centella against hydrogen peroxide was also studied in hippocampal cell line. These results were found to be reproducible in the other neuronal cell line PC12. Reactive oxygen species were found to be effectively reduced in in-situ as well as homogenized samples of H19-7 cells. Caspase activity was also found to be greatly reduced in presence of Centella extract. We also found Centella to increase the phosphorylation of ERK1/2 and CREB by itself at different doses and affect the NMDA receptors in differentiated and un-differentiated hippocampal neuronal cells. Similar observations were found when cells were treated with hydrogen peroxide in presence or absence of two concentrations of Centella. Thus, the current study describes anti-caspase, antioxidant actions of Centella namely attributing to its asiaticoside concentration. Modulation of glutamatergic receptors and stimulation of ERK1/2-CREB-BDNF pathway was found to be one of the putative mechanisms involved in synaptic strengthening shown by the Centella.
The impact of reduced SIRT3 in models of Neurodegenerative Diseases	Victoria S. Jiminez, Manoj Govindarajulu, Priyanka Das Pinky, Nila Ghanei, Vishnu Suppiramaniam and Rajesh Amin.	Dept of Drug Discovery and Development, Harrison School of Pharmacy, Auburn University,Auburn AL.	Alzheimer’s Disease is a neurodegenerative disease affecting an estimated 44 million people worldwide. Recent evidence has placed energy dysregulation at the focus for the progression from mild cognitive deficits to full dementia. Recently we observed a significant decrease in SIRT3 levels in advanced Alzheimer’s brains from 3xTG-AD, Diabetic (db/db) and SIRt3 KO mice. SIRT3 is a member of the sirtuin families of class III histone regulates mitochondrial protein function by deacetylation and thus is involved with metabolic homeostasis, oxidative stress, and cell survival. However currently there are no clinically available SIRT3 agonist for mitochondrial dysfunction associated with metabolic disorders. In the current study we have observed a significant decrease in long term potentiation when compared to age matched mice. Further analysis revealed significant hyperacetylation of key proteins involved in antioxidant capacity, including SOD. The current project proposal focuses on understanding the role of SIRT3 in the development of memory deficits as well as neuroinflammation. In addition we have observed impaired insulin signaling and reduced glucose uptake in SIRT3KO mice as well in 3xTG-AD and dbd/db mice brains. Lastly, we also observed enhanced neuroinflammation as demonstrated by elevated IBA1 expression in microglial cells. Together we observed reduced SIRT3 expression, reduced energy regulation and enhanced neuroinflammation in brains from SIRT3 knock out mice. Further these traits for neurodegeneration were similarly observed in aged 3xTg-AD and db/db mice. Further analysis will focus central mediators glucose uptake in these models of behavioral deficits and neurodegeneration.

Last modified: Jun 8, 2023 @ 11:44 am