Nutraceutical Effects on Delay Eyeblink Conditioning in Humans (Aging)

Small, Brent and Bickford, Paula

A considerable body of research indicates that multiple cognitive abilities decline as we grow older. As such, there is ever-increasing interest in the development of interventions to lessen or remediate age-related declines in cognitive performance among older adults. Evidence from epidemiologic studies, as well as experimental animal literature, suggests that dietary properties, especially antioxidant compounds, promote good cognitive health among older adults. In the current proposal, we examine the influence of a nutritional supplement intervention on the acquisition of delay and trace human eyeblink conditioning among older adults. Both delay and trace eyeblink conditioning are targetted because of their reliance upon different neural pathways, primarily cerebellar pathways for delay conditioning and medial temporal lobe pathways for trace conditioning. We predict that persons who receive the nutritional supplementation will exhibit facilitated eyeblink conditioning as compared to persons who do not receive the nutritional supplement. Moreover, we predict that measures of oxidative stress and inflammation will mediate nutrition-related gains in eyeblink conditioning. In order to test these predictions, we propose a randomized controlled trial whereby persons will be assigned to receive a nutritional supplement for a month preceding the eyeblink conditioning or receive a placebo during that period. In addition, participants will have blood drawn for the measurement of antioxidant markers and will also complete a brief battery of tests of memory and cognitive performance. Using these data, we will address the following specific aims: (1) Does nutritional supplementation facilitate acquisition in delay human eyeblink conditioning paradigm, relative to persons who do not receive this supplementation?; (2) Does nutritional supplementation facilitate acquisition in a trace human eyeblink conditioning paradigm, relative to persons who do not receive this supplementation?, and (3) Is the facilitation of the acquisition of delay and trace human eyeblink response in the nutritional supplementation group mediated by changes in anti-oxidant markers?

Heat Shock Protein Hsp27 as Regulator of Tau Fibril Assembly

Muschol, Martin and Dickey, Chad

Filamentous deposits of tau, a microtubule-associated protein, are either the primary molecular agent or significantly contribute to multiple senile dementias, such as Pick’s disease and Alzheimer’s disease. As a result, the cellular interactions of chaperone pro-teins with tau and its pathogenic variants have attracted considerable attention. This proposal will determine the mechanisms by which the molecular chaperone Hsp27 inhib-its tau fibril formation, and how various age-related mutations to either protein enable tau to evade and overwhelm cellular protein homeostasis. We have already developed re-combinant expression systems for human 4R0N tau, wild-type Hsp27 and a mock-phosphorylated mutant 3xS/D-Hsp27 and analyzed their biochemical interactions. We have also begun characterizing the in vitro kinetics of tau fibril-assembly, and have con-firmed that wtHsp27 does indeed suppress tau fibril growth.

To investigate the specific effects of Hsp27 on in vitro fibril assembly of tau we will utilize a multitude of biochemical and biophysical approaches including immuno-blotting and gel chromatography, sucrose-cushion ultracentrifugation, static and dynamic light scat-tering, CD spectroscopy and atomic force microscopy. Initially we will characterize tau fibril-assembly under near-physiological in vitro conditions, including quantification of tau net intermolecular interactions, detection of any transient intermediates and characteri-zation of their morphologies and dimensions, and determination of the nucleation and growth kinetics for each aggregate species. We will then determine whether and how interactions with the molecular chaperone Hsp27 alter each one of these distinct aspects of tau fibril-assembly. Finally, we will explore how age- and disease-related mutations to either protein disrupt the native interactions and accelerate subsequent fibril growth.

These experiments will interrogate the molecular processes regulating tau self-assembly, the mechanisms by which chaperones interfere with this process, and how age- and stress-related variants of tau or Hsp 27 disrupt native interactions. Further-more, these experiments will validate targets for subsequent animal studies and provide an in situ system for testing pharmaceutical agents to combat geriatric neurodegenera-tion.

Aerobic Exercise and Nicotine Withdrawal (Mood /Addiction)

Drobes, David and Kilpatrick, Marcus

Physical activity has been examined as a potential treatment adjunct for smoking cessation, but with mixed success and little clarity regarding mechanisms that might contribute to successful cessation outcomes. Ultimately, incorporation of exercise into smoking cessation treatment will benefit from a better understanding of specific effects of exercise on important aspects of smoking motivation, such as nicotine withdrawal. Previous research has demonstrated that nicotine withdrawal is associated with diminished executive attentional processes and mood deterioration, whereas aerobic exercise has positive effects within similar cognitive and affective domains. We propose that acute aerobic exercise may ameliorate cognitive and affective deficits during periods of nicotine abstinence. The direct effects of exercise on cognitive and affective functioning during nicotine withdrawal have not yet been systematically investigated. The primary aim of the proposed pilot research is to examine effects of acute aerobic exercise on measures of executive attentional cognition and mood during nicotine abstinence among thirty daily cigarette smokers. Using a 2 x 2 within subjects design, two of the sessions will occur in a state of nicotine deprivation, and the other two during a state of nicotine satiation. Participants will engage in aerobic exercise during one of each of these types of sessions, and a non-activity control during the other. Data analyses will focus on event-related brain potential (ERP) activity and behavioral performance during three well established executive function tasks (flankers, go-nogo, oddball), as a function of the exercise and nicotine abstinence manipulations. We will test the hypothesis that executive attention and mood indices will be diminished during nicotine abstinence, and that these deficits will be ameliorated following acute aerobic exercise. A secondary aim is to examine indices of tobacco use/dependence and initial fitness level (VO2Max) as potential moderators of the interactive effects of aerobic exercise and nicotine abstinence on executive attention and mood. Finally, genetic samples will be collected for future analyses. This research is meant to serve as the foundation for extramurally funded research concerning neurocognitive mechanisms that may underlie the effects of exercise on nicotine withdrawal, which may ultimately lead towards the optimal utilization of physical activity-based interventions for smoking cessation.

Robotics Modeling of Skilled Hand Tasks (Movement Disorders)

Lee, Sang-Hie and Sun, Yu

Hand motion of ten concert pianists will be recorded using Polaris Vicra System. Hand motion in piano practice of ten music students who have reported some level of hand discomfort will be recorded to create another data set. Data sets will be analyzed and compared. Motion features will be extracted and modeled to identify the prototype robotic model that represents a most proficient hand biomechanics and the principal biomechanical components that may contribute to hand injury. The components will be implemented on robot hands for verification and demonstration. Hand motion data of ten injured musicians will be compared with the healthy principal biomechanical components. We will then design an educational program for student pianists and a rehabilitation process for injured musicians. Functional dexterity of the robotic models will be tested on a MIDI (Music Instrument Digital Interface)-equipped piano to monitor the accuracy, key velocity, and timing of key depression and release. MIDI and motion analyses will be used to measure the outcomes of the programs.

This study aims to (1) capture skilled musicians’ hand motion in a robotic model to demonstrate efficient biomechanical movements that can be used in proficient music training and prevent hand injuries; (2) measure the principal hand biomechanical components of injured musicians with disabling symptoms, such as focal dystonia and design a rehabilitation process to bridge from injured hand motion to healthy hand motion; and (3) implicate the correct use of the hand for the larger population who use intensive repeated hand motion. The integrity of this study reinforces the USF’s Neuroscience’ strength by engaging in a new field of collaborative research and compliments the existing structures between the science and the arts.

This study, focused on the intensity of musician’s hand biomechanics, has a broader impact on school musician and teacher, popular and amateur musician, and other occupational hand users. It has a profound impact on music education since this will be the first time to introduce robot system in music learning. Non-musicians who use repeated complex hand tasks (e.g., computer) are often unaware of the biomechanically incorrect use, and many experience hand problems. This study is innovative, interdisciplinary, transformative, educational, and has broader impact. After dissemination of the study report, we hope to attract NSF, NIH, OSHA funding and industry partnerships.

Development of a Cubic Silicon Carbide Brain Machine Interface (PTSD/TBI)

Weeber, Edwin and Saddow, Stephen

When the central nervous system (CNS) is damaged through trauma or disease, gliosis repairs and regenerates some CNS tissue but also inhibits neural and axonal regeneration. One method of neural signal pathway restoration involves the use of a biomedical implant called the brain machine interface (BMI). This device has the ability to receive and transport signals between distant neurons and allow neuron interaction with computers and electromechanical prosthetics. The BMI can potentially restore functionality for people suffering from PTSD/ TBI, but they also may act as a neural pacemaker for Parkinson’s disease and depression and can signal muscle tissues for Muscular Dystrophy. Unfortunately, current material limitations facing the implantable component of the BMI blocks further progress and clinical trials: a lack of biocompatibility with the neural environment. Current BMI neural interfaces fail primarily due to their material’s eliciting secondary glial scaring that leads to eventual device encapsulation. BMIs also experience long-term material deterioration and biofouling due to the harsh body environment. We propose to use a novel material, silicon carbide (SiC), to construct a neural interface system for a BMI. SiC is chemically inert, physically durable, and has presented initial evidence of excellent in vitro biocompatibility.

The cubic polytype of SiC (3C-SiC) has displayed the highest in vitro biocompatibility, but a major critique of previously submitted federal grants has been the absence of true in vivo biocompatibility data. Specific Aim 1 of this proposal involves in vivo biocompatibility testing of 3C-SiC where we will compare CNS tissue reactions to implanted 3C-SiC shanks utilizing immunohistochemical staining, western blot proteomics, and microscopic investigation of the implanted material. We will construct a neuronal activation device (NAD) from 3C-SiC and explore its ability to receive and transmit signals from in vitro primary cultured neurons for Specific Aim 2. Specific Aim 3 will establish the ability of the NAD to elicit specific synaptic pathway excitation and perform signal integration with ex vivo organotypic hippocampal cultures. BMI devices have gathered much interest in the last decade due to many successful demonstrations with animal subjects, and the dual proof of biocompatibility of a 3C-SiC BMI device would position us well for garnering competitive external funding.

Optimal Timing and Dosing of Therapy for Cerebral Edema after Stroke (Stroke)

Decker, David and Pennypacker, Keith

Cerebral edema after stroke is associated with poor neurologic outcome and death. Current therapies are limited to osmotic agents, such as hypertonic saline (HS), which reduce intracranial pressure. Whether these therapies lead to increased tissue survival and the optimal method of administration is unclear. Additionally, newer agents that exploit different pathways, such as the vasopressin receptor antagonist conivaptan, have shown promise in preliminary studies, but have not been evaluated for synergy with osmotic agents. It is current clinical practice to delay treatment until patients become symptomatic from cerebral edema. Recent advances in the understanding of the pathophysiology of cerebral edema, however, suggest that early intervention may improve outcomes.

The specific aims of this project are to determine if HS and/or conivaptan reduce infarct volume and improve neurological outcome, and whether early initiation of therapy is superior to delayed initiation.

The long-term objectives are to develop a more effective treatment strategy for cerebral edema after stroke that will both reduce intracranial pressure and maximize tissue survival.

The design of this project involves the development of a standardized rat model for cerebral edema the may be used to explore different combinations of dosing, concentration and timing of initiation of HS and conivaptan and other agents in the future. Permanent middle cerebral artery occlusion will be used to induce ischemic strokes in rats. The animals will be treated with HS and/or conivaptan, beginning either 6 or 24 hours after infarction in varying dosages. After 48 hours, mortality and the degree of neurologic deficits in the survivors will be assessed. Brain tissue will be stained using a flouro-jade technique to assess infarct volume.

This project is likely to lead to sustained extramural funding as the results may be readily translated into human trials. Conivaptan is already FDA-approved for the treatment of hyponatremia, and if it shows a positive effect, the results of this study may eventually lead to an additional clinical indication for this medication. This animal model may also be used to test additional therapeutic agents and may be adapted for the treatment of cerebral edema found in other conditions including cerebral hemorrhage, traumatic brain injury and liver failure.

Genetic/epigenetic programming of human fetal brain by tobacco smoke

Salihu, Hamisu and Rubin, Lewis

Smoking accounts for a significant proportion of morbidity and mortality in the US with approximately half a million deaths attributed to smoking, and more than eight million people suffering from smoking-related illnesses. Despite smoking cessation programs, most smokers continue to smoke during pregnancy (prenatal smoking) leading to considerable fetal morbidity and mortality. An important consequence of prenatal smoking is its impact on the developing fetal brain leading to reduced intellectual capacity and subsequent adult-onset neurological disorders. Human studies on the mechanisms linking tobacco smoke to fetal brain damage are lacking limiting our understanding and ability to formulate targeted interventions early in life. Accordingly, we propose this study with the following specific aims: Specific Aim 1: To determine whether prenatal smoking reduces fetal brain size; Specific Aim 2: To identify and quantify alterations in brain regulatory gene expressions in cord blood of fetuses exposed to intra-uterine tobacco smoke; Specific Aim 3: To determine whether normal epigenetic modifications in specific genes responsible for normal fetal brain development are altered in fetuses exposed to intra-uterine tobacco smoke. The study will be conducted at the Tampa General Hospital delivery ward. Over the course of two years, we will recruit 140 pregnant women at delivery (70 smokers and 70 non-smokers). For all study participants and at the mother’s convenience, and preferably several hours after delivery but before hospital discharge, detailed questionnaires covering information on behavioral and nutritional habits, psycho-social stresses and mental health during pregnancy will be administered. Information pertaining to socio-demographic characteristics during pregnancy will be abstracted from the medical records. Neonatal cord blood will be collected at delivery for subsequent DNA extraction, genetic and epigenetic (DNA-methylation) analyses. The proposed project is multi-disciplinary and will provide critical information that will enhance our understanding of how tobacco smoke exposure modulates fetal brain programming during intra-uterine life. The study will also potentially provide compelling data for a subsequent R01 application to the NIH on the role of tobacco smoke in the fetal origins of childhood and adult-onset neuro-behavioral disorders.

Interactions of cdk7 spliced variants with PKC-iota during glioma cell cycle/proliferation

Patel, Niketa and Acevedo-duncan, Mildred

The broad, long-term goal of this proposal is to design better treatments for glioma. High-grade gliomas are highly lethal brain tumors that arise from the supportive glial/neuroglial tissue. We seek to understand the interaction between protein kinase C-iota (PKC-i or PKC-ι) and the master cell cycle regulator, cyclin-dependent kinase activating kinase (CAK/Cdk7) during the glioma cell cycle and proliferation. This line of inquiry follows from our results showing that PKC-ι is associated with two forms of CAK/Cdk7 (Cdk7_L (42kD) and Cdk7_S (37 kD)) which are temporally regulated throughout the cell cycle. This project will examine whether cell cycle progression in gliomas can be regulated by the interaction of PKC-ι and Cdk7 splice variants. Our hypothesis is that Cdk7_S splice variant functions as a repressor of G1/S transition.The specific aims are : 1. Determine the status of Cdk7_L and Cdk7_S in normal, benign and malignant brain tissue. 2. Establish the mechanism by which Cdk7_S is regulated 3. Determine a role for Cdk7_S in G1/S transition or gene transcription 4. Determine whether Cdk7_S modulation affects cytokinesis or DNA synthesis 5. Elucidate alternative splicing mechanism resulting in Cdk7_L and Cdk7_S splice variants. The proposed research will be performed in normal, benign and malignant brain tissue and in-vitro in control normal human astrocyte cells (NHA) to evaluate malignant progression, in the U-138 glioma cell line and in NHA Cdk7_S sense overexpressor cells in order to compare results. The experimental methods are 1) cell biology techniques; cell cycle analysis, flow cytometry, proliferation assays; 2) biochemical procedures; cell subfraction, PKC activity assays, Western blotting, and 3) molecular biology techniques and splicing assays. These studies are relevant because elucidation of mechanisms by which Cdk7_S regulates the cell cycle and proliferation may suggest ways to mimic it role in the cell, thereby providing selective therapeutic agents for controlling glioma proliferation. Knowledge on how Cdk7 splice variants may control the defects which control the unrestricted proliferation of gliomas relates to the USF NSC program area by fulfilling the requirements of shared criteria for secondary clusters. Drs. Patel and Acevedo-Duncan have never collaborated and are in different departments at USF. This project will provide the needed preliminary data to apply for sustained external funding from NIH (RO1) and NSF.

Symposium on Early Alzheimer's Disease

Potter, Huntington and Duara, Ranjan

The purpose of this symposium (formerly the Mild Cognitive Impairment Symposium) is to provide a forum for new information and for in-depth discussions about advances in research, related to the clinical diagnosis, progression and treatment of Alzheimer’s Disease. This symposium will emphasize the early stage of AD and the preclinical deficits that are associated with increased risk for subsequent dementia. We will examine recent developments in understanding risk factors which lay the groundwork for potential strategies for treating and delaying Alzheimer’s disease. We will present results from several longitudinal and cross-sectional studies that have explored the relationship between clinical, imaging and biomarker results and AD.

Establishing Autism Spectrum Disorder Networks

Mieres, Ana and Vaughn, Bobbie

The conference, an interactive symposium, will bring national experts in the field of autism from different disciplines and professions within USF to present advances in Autism Spectrum Disorders (ASD) to 100 selected participants working within the field of ASD. Participants will engage in small interprofessional group discussions and problem solving sessions, to assess and evaluate full session information and examine the translational capabilities for education, research and clinical practice locally, regionally and nationally.

Professionals working within the field of ASD to include: physicians, psychologists, nurses, physician assistants, social workers, physical therapists, speech and language pathologists, occupational therapists, applied behavior analysts, special education teachers, administrators, third party payer case managers and other advocacy groups are asked to join to build competency , understand the roles of others in the continuum of care, and jointly assess the new advances for families with children diagnosed or suspected of having autism spectrum disorder.

Invited participants will respond in advance to invitations to allow the structuring of assigned table seating (active learning pods), among different professions. A trained facilitator and a trained note taker will accompany each group to administer and facilitate discussion on pre-developed questions. The group facilitation will occur after each formal plenary session. The questions posed to the groups will serve to evaluate, expand and advance the presented information and systematic facilitation will occur resulting in information that will impact pre-service education, continuing education, research and clinical practice. Reports from each table will be compiled for a Symposium report.

The proposed Symposium will replicate the Ohio State University, Commission for Interdisciplinary, Education and Practice (CIEP) Conferences for both the National Consortium and the Ohio State Commission.

Anti-apoptotic effects of nicotinic agonists

Wecker, Lynn and Citron, Bruce

The goal of this research project is to identify the cellular and molecular targets and mechanisms mediating the ability of neuronal nicotinic receptor modulation to alleviate the incoordination and imbalance in individuals with ataxia, thereby leading to the development of pharmacological agents to treat these disorders. There is no current pharmacological treatment to alleviate the imbalance and lack of voluntary muscle coordination manifest by individuals with ataxia. Ataxia may be a consequence of hereditary diseases or non-hereditary causes including chemical or traumatic insult, and is ascribed typically to a loss of function in the cerebellum or its associated afferent or efferent pathways. Clinical studies have indicated that varenicline, a partial agonist at α4β2 and full agonist at α7 neuronal nicotinic receptors, improves balance and coordination in patients with ataxia of distinct pathogenic etiologies including disorders that are a consequence of diverse molecular and cellular alterations and involve different anatomical pathologies. Preliminary studies in our laboratory have provided proof of concept for the use of nicotinic agonists in an animal model of human olivocerebellar degeneration, but the mechanisms involved are unknown. This translational project will use a classical pharmacological approach to determine whether the ability of nicotinic agonists to alleviate ataxia in this laboratory rat model is a consequence of the selective activation of α4β2 or α7-containing neuronal nicotinic receptors, whether receptor activation shifts the balance between pro-apoptotic and anti-apoptotic signaling in the cerebellum to favor the latter, and whether receptor activation increases the expression or levels of IGF-1, a growth factor critical for the integrity of cerebellar neurons and implicated to play a pivotal role in human cerebellar degenerative disorders. Results will provide data necessary for the preparation of a major extramural grant proposal and lead to the development of new therapeutic agents for the treatment of these disorders for which there is no current efficacious pharmacological therapy.

Subtypes of callous-unemotional traits in "cross-over" youth

Kimonis, Eva and Groer, Maureen

The long-term objective of this proposal is to test whether there are two trajectories to callous-unemotional (CU) traits (lack of guilt, empathy, callous use of others) in youth that are underpinned by distinct emotional profiles. This study will generate basic behavioral and neuroscience findings that can be directly translated to inform the assessment and treatment of antisocial behavior in at-risk youth. It is well-established that a subset of youth with more severe, persistent, and early-onset conduct problems shows CU traits. Prior research consistently documents that these youth show low autonomic reactivity and deficient processing of negatively valenced emotional stimuli. However, youths with manifest CU traits may be a heterogeneous group. Specifically, recent research shows that those with comorbid psychopathology (anxiety, depression), along with a history of trauma, show the opposite pattern of heightened emotional reactivity. Theory suggests that behaviorally observed emotional reactions following trauma exposure can resemble CU traits (secondary type). The proposed cross-sectional study will test whether incarcerated delinquent youth high on CU traits, but grouped into primary and secondary types according to maltreatment history and co-occurring psychopathology, show distinct neuropsychological, psychophysiological, and neuroendocrine profiles. Official administrative data will be combined with youths’ self-reports to determine maltreatment history. Youth will complete a comprehensive emotional assessment, including neurocognitive and psychophysiological testing, and measurement of stress hormone levels. These data will be used to support larger collaborative R01 federal grant applications to test, using longitudinal designs, developmental trajectories to CU traits in youth, and to test the efficacy of comprehensive and individualized interventions for antisocial and aggressive youth.

The proposed project has two specific aims:

Aim 1: Test whether primary and secondary subtypes of CU traits, as distinguished by maltreatment history and psychopathology, reflect distinct neuropsychological, psychophysiological, and neuroendocrine profiles derived from a multi-method assessment of emotion.

Aim 2: Test whether youths exhibiting CU traits and a maltreatment history (secondary type CU) have greater PTSD symptoms and substance abuse/dependence than primary-subtype CU youths (low psychopathology, trauma history) and youth low on CU traits.