Disruption of ion channel function is increasingly associated with neurodevelopmental disorders (NDDs) such as autism spectrum disorder (ASD). ASD has complex etiology and symptoms, but recent studies suggest that individuals with ASD have disrupted mesostriatal network activity associated with social interaction deficits. Although mutated ion channels play a key role in NDDs, the degree to which they disrupt mesostriatal network activity is not well understood. The Kv7 family of voltage-gated potassium channels encoded by KCNQ1-5 genes are increasingly linked to NDDs including ASD. These channels are broadly expressed, including within the ventral tegmental area (VTA), a nucleus of the mesostriatal pathway that plays an important role in social behavior. Here we assessed the impact of the ASD-associated variant KCNQ3(R2C) on VTA dopamine neuron excitability and social behavior in mice. We used a viral-based strategy to conditionally inactivate endogenous Kv7.3 with CRISPR/SaCas9 and re-express human wildtype (hKv7.3/WT) or mutant (hKv7.3/R2C) KCNQ3. Whole-cell patch clamp electrophysiology in brain slices revealed that hKv7.3/R2C significantly decreased excitability of dopamine neurons. I also observed a loss of social novelty preference using the three-chamber social interaction task in mice expressing hKv7.3/R2C but not in controls expressing hKv7.3/WT. These results suggest that hKv7.3/R2C and other similar Kv7 mutant variants may disrupt mesostriatal network activity and social interaction by decreasing dopamine neuron excitability.

In 2020 marijuana use among college students was at an all-time high with 44% reporting using in the past year. Research demonstrates individuals with ADHD are at risk for increased marijuana use later in life. Additionally, some studies demonstrate self-reported symptoms of ADHD among undiagnosed college students prospectively relate to increased marijuana use, which was mediated by changes in perceptions of typical peer marijuana use (i.e., descriptive norms). However, this is complicated by frequent marijuana use being linked to issues with attention, impulsivity, and memory; all captured by self-reported ADHD measures. This study will evaluate bidirectional relations between ADHD-related symptoms, marijuana descriptive norms, and marijuana use among college students. Data collection is in progress and will be completed in March 2022 as part of a larger study. Students completed assessments at baseline, 6 months, and 12 months. A cross-lagged panel model will be conducted in R using the lavaan package to evaluate the bidirectional associations over a 1-year period, as well as test whether marijuana norms at 6 months mediate the longitudinal associations between ADHD-related symptoms and marijuana use from baseline to 12 months.  We hypothesize those reporting more ADHD-related symptoms will use marijuana more frequently 6 months later. Additionally, those who use marijuana more frequently will report more ADHD-related symptoms after 6 months. We also hypothesize that those experiencing more ADHD-related symptoms at baseline will have increased marijuana norms at 6 months and subsequent greater marijuana use at 12 months. This study attempts to disentangle the bidirectional associations between ADHD-related symptoms, marijuana norms, and marijuana use over a one-year period. Conclusions and future directions for research will be presented. Additionally, this study may have implications for norms-based interventions to better address memory, impulsivity, and other ADHD-related symptoms in association with marijuana use and perceptions.

Despite the well-known role of the neurotransmitter dopamine in reinforcement learning, generating the brain’s patterns of dopamine release for conditioned response learning remains unresolved. The Zweifel laboratory discovered that two ion channel subunits, Kv4.3 and BKCa1.1, expressed in ventral tegmental area (VTA) dopamine-producing neurons control the pattern of dopamine neuron firing at the cell body and dopamine release in the nucleus accumbens (NAc) on different time scales to regulate separate phases of reinforced behavior in awake and behaving mice (in vivo). However, whether the impact on dopamine release is dependent on cell body loss of function (LOF) or possible effects of local terminal regulation remains unknown. DLight—a fluorescent, genetically modified dopamine receptor—was injected into the NAc. Chrimson, a photoactivatable ion channel, and CRISPR/Cas9 viruses that knocked out either of the two ion channel subunits were injected into the VTA for selective expression in dopamine neurons. Three conditions were imposed: control (no subunit knockout), Kv4.3 LOF, or BKCa1.1 LOF. After at least six weeks, NAc brain slices were prepared for two-photon imaging of dLight following photoactivation of VTA dopamine terminals. I measured changes in dLight fluorescence, indicating dopamine release, using ImageJ and analyzed them using ImageJ, Excel, and GraphPad Prism. VTA dopamine terminals in the dLight-expressing NAc of control and Kv4.3 LOF mice were photostimulated; the BKCa1.1 LOF mice are in progress. I found that loss of Kv4.3 in VTA dopamine neurons does not impact dopamine release ex vivo. Observed differences in dopamine release in vivo in Kv4.3 LOF mice and lack of differences in ex vivo suggest that the ion channel effects at the cell body impact dopamine release downstream, highlighting that regulators of dopamine neuron cell body activity patterns impact dopamine release and affect dopamine-mediated behaviors like learning and motivation.

The ongoing opioid epidemic has made apparent the consequences of repeat opioid use and the opioid withdrawal that follows. Avoidance of opioid withdrawal and its associated symptoms has led to a need to understand signaling within neural circuits involved. The parabrachial nucleus (PBN) is a region in the brainstem that relays sensory information throughout critical forebrain regions, informing specific behavioral actions and learning mechanisms. A genetically defined cell population with the PBN expressing the calcitonin gene related peptide (CGRP, gene name Calca) has been demonstrated to relay aversive sensory and visceral information when activated. I hypothesized that PBN-CGRP neurons may be a neuron population that is recruited during opioid withdrawal. To investigate whether the aversive experience of opioid withdrawal recruits PBN-CGRP neuron activity, I induced morphine-dependence in mice expressing a yellow fluorescent protein in PBN-CGRP neurons. After establishing morphine dependence, precipitated opioid withdrawal was induced with an injection of naloxone. Ninety minutes after induction of precipitated opioid withdrawal, mice were perfused to fix brain tissue. I then performed immunohistochemistry for cFos, an immediate early gene and a marker of increased neural activity, in brain sections with the PBN. I am quantifying the expression of cFos, a marker of increased neural activity, in PBN neurons that contain the neuropeptide CGRP following naloxone precipitated withdrawal. I expect to see increased cFos expression indicating the recruitment of PBN-CGRP neurons in mice that are suffering from opiod withdrawal compared to the control mice. By understanding whether the PBN is recruited during opioid withdrawal, I can begin to explore options to alter neuronal activity in the PBN to lessen the severity of symptoms felt during opioid withdrawal, a key contributor to the prevalence of relapse in those overusing opiates.

Alcohol withdrawal exists as a significant barrier to care for those who struggle with alcohol use disorder. Furthermore, there is evidence of worsening withdrawal symptoms after repetitive cycles of alcohol use and abstinence. Our preliminary research points to neuroinflammation primarily regulated by microglia, the immune cells in the brain, as a potential causal component in the escalation of recurring alcohol withdrawal symptoms. Previous work in our lab has shown that mice subjected to chronic intermittent exposure (CIE) to ethanol vapor showed elevated markers of neuroinflammation, as evidenced by transcriptome analysis in microglia. We found that the “unfolded protein response (UPR)” was activated; it plays a critical role in the development of neuroinflammation, and the UPR is mediated significantly by the C/EBP homologous protein (CHOP) an apoptotic transcription factor. Therefore, we hypothesize that CHOP in microglia is necessary for promoting alcohol withdrawal symptoms. We expect to observe measurable reductions in the withdrawal symptoms experienced by transgenic male and female mice with and without CHOP following CIE exposure. In our pilot study, transgenic male and female mice with and without CHOP were subjected to five weeks of CIE to ethanol vapor and then tested for withdrawal phenotypes. These tests include body temperature, locomotion, marble-burying behavior, novelty suppressed feeding, and sucrose preference. We have also characterized microglia morphology and CHOP expression in pilot mice brains. Having measured changes in these mice's neuroinflammatory mechanisms and subsequent behaviors, our results will determine the efficacy of decreasing CHOP gene expression to ameliorate the kindling effect in alcoholics. Therefore, our work provides a promising direction in tackling the inescapable cycle of worsening alcohol withdrawal that limits the road to recovery for alcoholics.

Autism Spectrum Disorder (ASD) is a developmental disorder that can impair social and communication skills. Many individuals diagnosed with ASD experience other mental health disorders such as anxiety and depression. Research has estimated that between 63-70% of people with ASD suffer from depression while 50-58% experience anxiety. Anxiety and depression both have been found to have a negative correlation with self-compassion for ASD children and typically developing (TD) children. Self-compassion is the concept of being caring towards oneself and is characterized by three facets: (1) self-kindness (2) common humanity, and (3) mindfulness. The present study seeks to explore the relationship between self-compassion and anxiety/depression and how this may vary for children with ASD compared to TD children. To examine this, 55 children (ASD=21), ages 7-17 years from the longitudinal ACE2 study, a five-site NIH-funded study on gender differences in autism were included in the analysis. ASD diagnosis was confirmed via standardized measures. Self-compassion was measured using the Self-Compassion Inventory for Youth, a self-report tool that generates scores on two scales: self-compassionate coping scale and self-punitive coping scale. Anxiety and depression were measured using the Youth Self Report (YSR), a self-questionnaire assessing symptoms of anxiety and depression. We expect that children with ASD will experience a stronger negative correlation between self-compassion and anxiety/depression than TD children. This relationship is critical to understand as individuals with ASD can experience difficulty practicing self-compassion. Cognizance of the relationship between these two constructs may help inform beneficial interventions and treatments for children with ASD to alleviate symptoms of anxiety and depression.

Drug addiction and abuse exacerbate many health issues globally. The nucleus accumbens (NAc), a region within the brain, contains dopamine receptor D1 and D2 medium spiny neurons (MSNs) with relevance to addiction-related behaviors. I investigated the effect of D2 MSNs on behavioral economics of cocaine-seeking through chemogenetic activation of Designer Receptors Exclusively Activated by Designer Drugs (DREADD). I used a transgenic line of rats specifically expressing the Cre-recombinase enzyme in D2 MSNs. This enzyme is responsible for selective expression of otherwise-inert viral vectors by reversing its sequence, allowing for neuron-specific genetic manipulation. I bilaterally injected either a Cre-dependent AAV containing the DREADD hM3Dq or a fluorescent marker as control into the NAc of male rats; additionally, I catheterized the rats to allow for cocaine self-administration. Activation of hM3Dq required binding to clozapine-N-oxide (CNO)—the “designer drug”—and because only D2 MSNs expressed this DREADD, they were the only neurons to activate upon administration of CNO. After recovery, rats were trained to press a lever to receive cocaine infusions; and once this self-administration task was learned, the animals were introduced to reinforcement schedules with blocks of trials with differing unit doses of cocaine. Once this phase was learned, the rats were tested in sessions that were preceded either with an injection of saline or CNO, to test how chemogenetic activation of D2 MSNs affects the cocaine demand curve. Based on studies discussing the roles of D2 MSNs, I predict that chemogenetic activation of these neurons will correlate with decreased demand for cocaine concentrations compared to those injected with vehicle or green fluorescent protein alone. This study can produce valuable insight into the role this pathway plays in the development of drug-seeking behavior, ultimately leading to a deeper understanding of the system and possible avenues for treating addiction.

Soldiers who are transitioning to civilian life have alarmingly high rates of suicide, alcohol abuse, and mental illness. There are multiple facets that may influence these poor outcomes for veterans. Organizational dissatisfaction is a risk factor for poor mental health and lack of access to mental health support for veterans, but this has yet to be studied in-depth. The aim of this project is to assess how demographic variables and organizational dynamics impact mental health during the transition process. First, we will use preliminary data from The Network Study, one of the first longitudinal studies to follow the mental health, alcohol use, and socioeconomic stressors of service members as they transition out of the Army. Multiple regression will be used to analyze the influence of interpersonal support and conflict within the Army on depressive and anxiety symptoms. Mental health symptoms were measured by the PHQ-9 and GAD-7, two common screening questionnaires used to briefly assess depressive and anxiety symptoms, respectively. We will additionally assess various demographic variables as moderators. Next, findings will be used to develop questions for qualitative interviews to be conducted with recent veterans. We predict there will be a positive association between organizational support and positive mental health outcomes, and qualitative interviews will show lower work-related stress and increased access to mental health supports as major themes for those with positive work relationships. In line with previous research, we predict these associations and themes will be strongest for women, racial and ethnic minorities, and those of lower rank. Support and intraunit conflict are potentially modifiable factors, so our research demonstrates a potential pathway for prevention of negative mental health outcomes for veterans.

Engagement barriers are barriers that prevent individuals from participating in Attention Deficit Hyperactivity Disorder (ADHD) therapy. Kazdin et al. (1997) found that having fewer engagement barriers serves as a protective factor to prevent treatment dropout. Despite knowledge behind the importance of having few engagement barriers, who experiences what barriers is largely unexplored. By understanding what engagement barriers can arise during behavioral therapy, and who experiences them, therapists can generate tools to overcome these barriers, thus improving the treatment of adolescent ADHD. Our purpose of this study was to ascertain and understand any relationships between engagement barriers in behavioral therapy and clinical characteristics/sociodemographic factors. Participants were 121 adolescents with ADHD (which came from an ethnically diverse sample) and their parents. Twenty-five different barriers were sorted across three subtypes (cognitive/attitudinal, behavioral, and logistical), three sources (parent, teen, or dyadic) and three treatment phases (engagement, skills, and planning). Minority status, gender, ADHD medication status, family adversity, Oppositional Defiant Disorder (ODD) comorbidity, and parental ADHD served as independent variables. I used multiple regression models to analyze relationships between each independent variable and barrier subtype, source, and treatment phase. ODD comorbidity and parental ADHD each significantly predicted a primary outcome of an overall increase in barrier frequency. Significant secondary outcomes included ODD comorbidity and parental ADHD each significantly predicting greater cognitive barriers, teen barriers, engagement phase, and skills phase barriers frequencies. ODD also predicted significant increases in planning phase barriers frequencies, while minority status significantly predicted lower planning phase barriers frequencies. These results demonstrate that populations of adolescents diagnosed with ADHD and comorbid ODD or who have parents that are also diagnosed with ADHD are at risk to experiencing more engagement barriers. Additional research is required to develop therapeutic tools. These tools will help therapists better serve adolescents with comorbid ADHD/ODD, and parents with ADHD.

Chronic sleep disruption, present in 25-60% of patients suffering from Alzheimer's Disease (AD), often precedes cardinal disease symptoms. While little is known about the mechanisms underlying chronic sleep disruption and the development of clinical pathology, both acute and chronic sleep deprivation have been found to increase biomarkers of AD including neuroinflammation and amyloid-beta accumulation. Additionally, in people without AD, sleep deprivation can result in a deterioration of working memory and attention. In this study, we examine the impact of chronic sleep disruption on cognition both at baseline and in the 5xFAD mouse model of AD. The 5xFAD mouse model is a transgenic mouse model of familial amyloidosis which expresses neurocognitive impairment as early as 2 months. To define the effect of chronic sleep disruption on cognition in the absence of AD pathology, 60 wild type mice were exposed to chronic sleep disruption or sham procedure for 8 weeks between 10 and 18 weeks of age. At 18 weeks of age, I evaluated the animals for changes in spatial memory (Barnes maze), short-term memory (Y-maze), locomotion and anxiety (open field test), and activities of daily living (burrowing trials). To test whether chronic sleep disruption specifically exacerbates AD-related neurocognitive decline, the same cognitive tests were performed on 60 5xFAD+ animals exposed to 8 weeks of sleep disruption or sham treatment. I then analyzed the collected data to isolate any trends of cognitive performance, finding that chronic sleep disruption impaired cognitive performance in 5xFAD+ and littermate controls, with a more significant impact on 5xFAD+ animals. These findings highlight the critical association between dysfunctional sleep and the development of cognitive impairment with AD disease progression which then guides us toward better preventative care and treatments.

Mild traumatic brain injury (mTBI) is a major public health issue, frequently resulting in long-term sequelae such as sleep disruption, headaches, and cognitive impacts. In recent years, mTBI has emerged as a risk factor for the development of neurodegenerative diseases, such as Alzheimer’s disease (AD). Blast-related mTBI has been experienced by large numbers of Servicemembers during the conflicts in Afghanistan and Iraq; therefore, their potential vulnerability to downstream neurodegeneration is a major concern among Veteran populations. Recent evidence demonstrates that TBI impairs the glymphatic system, a brain-wide network of perivascular channels along which cerebrospinal fluid (CSF) and interstitial fluid (ISF) exchange facilitates the clearance of interstitial solutes such as amyloid β and tau. However, these findings were in an impact TBI model, which is a brain injury caused by a blow to the head; therefore, relatively little is known about the possible effects of blast mTBI. Here, we hypothesize that glymphatic function is impaired following repetitive blast mTBI. Using a murine blast model, we measured glymphatic function at both 7-day and 28-day timepoints following a repetitive blast induced TBI. Glymphatic function was quantified using intracisternal fluorescent tracer injection and measuring the fluorescent intensity of CSF tracer movement. We found a delayed impairment in glymphatic function at 28 days post-injury. These findings may provide further insight on the mechanisms that may render the blast-injured brain vulnerable to neurodegeneration and may give rise to improved treatments for patients exposed to blast injury.

Alzheimer’s disease (AD) is an age-related neurodegenerative disease characterized histopathologically by amyloid plaques and neurofibrillary tangles in the brains of affected individuals. The impairment of cerebrospinal fluid (CSF)-mediated clearance of proteins including amyloid beta and tau from the brain is proposed to underlie the development of AD pathology. Sleep and circadian disruption are both linked to the development of AD. CSF clearance is regulated through both sleep and circadian processes, while CSF production by the choroid plexus (CP) is diurnally regulated. The CP acts as a blood-CSF-barrier, provides nutrient delivery, and clears toxic proteins. Studies to date document reduction in CSF production, impaired blood-CSF-barrier function, and altered protein uptake in both aged and AD conditions. It is currently unknown whether the CP is itself regulated by sleep and circadian rhythms and whether disruption of these two governing processes contributes to disease development. Our initial analysis across awake, asleep, and acutely sleep-deprived young mice indicated no gene expression differences between sleep states; however, significant circadian-dependent transcriptional changes were observed. We then examined the circadian-dependent gene expression profile of the CP in aged (12-14 months) mice and in an AD mouse model. Preliminary analysis reveals a shift in the transcriptional profile of aged mice and a near complete loss of circadian regulation in the AD model. Ongoing analysis and validation are being carried out to reveal functional pathways disrupted in the CP that could provide a further understanding of AD pathology.

The glymphatic system, which is primarily active during sleep, is a network of astroglial perivascular channels within the brain that allow for Cerebrospinal Fluid (CSF) influx and exchange. Glymphatics play a crucial role in the waste clearance of amyloid beta, a hallmark in the development of Alzheimer’s Disease and neurodegeneration. Recently, a bidirectional relationship between Alzheimer's Disease and sleep has also been suggested with the aggregation of amyloid beta associated with mid-life sleep disruption. However, the mechanistic link between sleep disruption, particularly over chronic time scales, and the development of Alzheimer’s pathology remains unclear. This study investigates whether chronic sleep disruption, similar to that experienced in humans, will impact downstream neuropathology. We hypothesize chronic sleep disruption will result in decreased glymphatic function and subsequently increased amyloid plaque burden. This experiment utilizes a chronic sleep fragmentation model in 120 5xFAD mice from 8 weeks to 16 weeks of age. In the Lafayette Sleep Fragmentation chambers, 60 animals are disturbed every two minutes during normal sleeping periods (daylight hours). 60 mice were placed in normal sleeping conditions. After eight weeks of sleep fragmentation or sham exposure, glymphatic function is assessed by in vivo near infrared imaging following stereotactic CSF tracer injection. Animals are perfusion fixed, cryosectioned, and glymphatic function is assessed by measurement of fluorescent cerebrospinal fluid tracers in brain tissue. Aquaporin-4 localization, amyloid plaque deposition, and markers of astroglial and microglial activation are assessed by immunofluorescence. In this project, I specifically work on cryosectioning the tissue, and understanding glymphatic function through the processes of immunofluorescence imaging and analysis. The collected data demonstrated that sleep disruption did increase neuropathological outcomes.The measured impact of glymphatic function was also correlated with these downstream pathological effects. These findings could be an indicator of interactions between neurological disease progression and an inflammatory expression after sleep disruption. They can also shed more light on the complex relationship between Alzheimer’s disease progression, the glymphatic system, and chronic sleep disruption.

Alzheimer’s Disease (AD) is a neurodegenerative disease that affects more than 5 million Americans. The glymphatic system (a network of perivascular spaces that facilitate fluid movement and solute clearance from the brain) and its dysfunction associated with aging has been implicated in the development of AD. The water channel aquaporin 4 (AQP4), located in astrocytic endfeet bordering the perivascular spaces, is crucial for the proper functioning of the glymphatic system. Data suggests that loss of AQP4 localization results in amyloid-ß deposition, a hallmark of AD pathology, and loss of AQP4 localization accompanies aging in rodents as well as AD in humans. In this study, we quantitatively analyze the expression of aquaporin- 4ex (AQP4ex)—a translational readthrough variant of AQP4 believed to play a role in its localization—to identify any correlation with aging and AD pathology. Selective deletion of AQP4ex results in the mislocalization of AQP4 all over the astrocytic membrane, indicating that AQP4ex is a crucial element in the localization of AQP4. We analyze young, old and AD groups in the murine (mouse) brain as well as AD versus control in a human case series. Currently, we see a trend towards decline in cortical perivascular AQP4ex in the AD group, with more analysis ongoing. This is the first characterization of AQP4ex expression in the murine brain and in a human case series, and these data will contribute to the small but growing body of research on AQP4ex and its relationship with AQP4 localization, creating opportunities to identify a new novel mechanism and novel target in AD pathology.

Sexual minority women, and bisexual women in particular, are at greater risk of experiencing sexual assault and other types of traumas and stress, as well as heightened symptom severity than heterosexual women. Among sexual minority women, bisexual women in comparison to lesbian women have shown a higher risk of reporting worse mental health. To our knowledge, only three studies have looked at differences in mental health in bisexual and lesbian women who have been sexually assaulted, but their results have been contradictory. As such, the purpose of this study is to investigate the symptom differences between bisexual and lesbian women following sexual assault to better understand their differences. To do this, I conducted a secondary analysis of an existing national data set that surveyed 1057 sexual minority women. I used bivariate statistics tests to compare lesbian and bisexual women regarding their mental health following sexual assault with a focus on anxiety, depression, and PTSD symptoms. No statistically significant differences were found between the two sexual identity groups as a function of sexual assault, but bisexual women reported greater symptoms of depression regardless of sexual assault status in comparison to lesbian women. A potential explanation for this difference could be that bisexual women may have higher levels of minority stress (i.e., stress resulting from societal prejudice and other stigmatizing experiences) than lesbian women, which has been connected to heightened mental health issues. Future studies should consider minority stress as a factor when studying the two sexual identities to better tailor research and treatment to bisexual and lesbian women’s unique identities.

STAND (Supporting Teens Autonomy Daily) is an evidence-based practice modular therapy that helps ADHD (Attention deficit hyperactivity disorder) teens build academic and interpersonal skills. STAND has been shown to be cost-effective in promoting evidence-based practices compared to the usual-care psychological services and medication for ADHD. The purpose of this study is to assess the utilization and efficacy of online resources and training tools for therapists and supervisors implementing STAND. Six therapists and four supervisors administered STAND with the use of LYSSN, an AI (Artificial Intelligence) based tool that provides feedback on therapy audio tapes, and Care4, a dashboard that provides supervisors with a library of resources. I analyze 49 therapist sessions and 3 supervision sessions to determine the attitudes and engagement of the therapists and clients with STAND activities by calculating the average of scores provided by therapists and supervisors from Care4. Therapists login onto the Care4 system after a therapy session and answer questions about feedback they received from LYSSN and the preparation for their session. Supervisors used the data collected from the therapist’s LYSSN score and formulate their feedback for therapists. Therapists spent an average time of 11 to 30 minutes reading through the STAND session description and preparing client worksheets and an average of 1 to 10 minutes reviewing LYSSN feedback, watching STAND demonstration videos, seeking consultation from the STAND research team, and reviewing feedback from their supervisor. Therapists found LYSSN to be “occasionally” accurate and credible. Collection of this research will provide insight about the implementation of STAND through an online format and about the general appeal of the use of AI, the workload of supervisors and therapists could be reduced and give feedback more efficiently to them with regards to STAND treatment and other evidence-based treatments.

The adolescent age is ideal for Attention-Deficit/Hyperactivity Disorder (ADHD) treatments, such as Supporting Teen's Autonomy Daily (STAND) which was developed to make therapy accessible to disadvantaged youth with ADHD, since these treatments may be carried from the teenage years into adulthood. To increase the quality of care for these adolescents and improve their outcomes, six therapists and four supervisors have received STAND training. This training uses technology, such as the Artificial Intelligence (AI) tool that gives them feedback: Lyssn, is used to enhance provider engagement and delivery. The purpose of this study is to examine whether this STAND training and its technology are effective in improving therapists' STAND delivery methods to patients. Through 64 previously coded audio recordings of provider sessions that took place in Spring and Summer 2021, I assess the effectiveness of this AI training for therapists and their supervisors on their performance and delivery of STAND treatments. To do this, I analyze the Motivational Interviewing Treatment Integrity (MITI) coding system scores that the therapists received as feedback from Lyssn, when the therapy sessions occured, when those sessions were uploaded, and when therapists received feedback. I also examine questionnaires given to therapists regarding their thoughts on Lyssn before and after their sessions. Though data is preliminary and lacks a control group, results could inform us of therapists' positive engagement with Lyssn. Using AI feedback to train therapists on new practices or more efficient ways to deliver ADHD therapy sessions could potentially produce significant benefits to disadvantaged adolescents with ADHD so that they receive optimal care and see positive outcomes.

The nucleus accumbens core (NAc), among other brain regions, plays a key role in drug seeking behavior and relapse, particularly in determining incentive value as cocaine consumption escalates. Neurons in this region have two genetically-distinct output projections which form the direct and indirect pathways, D1 and D2 respectively. My experiment is based on the Incubation of Craving model, in which animals show increased drug seeking after a period of abstinence from self-administered cocaine. The behavioral side of this project seeks to better understand how these two pathways contribute to drug seeking behavior following a period of forced abstinence, and I am also investigating how RNA translation changes in and between the direct and indirect pathways following the escalation of cocaine taking and incubation of craving by forced abstinence. I used a transgenic line of rats expressing the Cre-Recombinase enzyme in the D1 and D2 medium spiny neurons (MSNs) to selectively express and manipulate MSNs in the direct and indirect pathways. I bilaterally injected either DIO-hM4Di RiboTag, DIO-hM3Dq RiboTag or a fluorescent control virus into the NAc, and the rats were catheterized during this time to allow for cocaine self-administration. The rats underwent the Incubation of Craving experiment where they experienced an acquisition and abstinence period. Once the Incubation of Craving was complete, I collected and homogenized the NAc from each subject and performed RNA purification. Then, I performed qPCR and RNA sequencing to investigate, and validate any pathway-specific changes in mRNA expression following these behaviors with the goal to discover new therapeutic targets. My hypothesis is that ribosome-associated mRNA in synaptosomes differs from the cell bodies, with enrichment of RNAs known to be trafficked to dendrites. Additionally, I predict that incubation of craving will induce distinct patterns of RNA changes in neurons of the direct and indirect pathway.

Traumatic brain injuries (TBIs) are a major cause of disability among war veterans, leading to behavioral dysfunction and post-concussive symptoms such as depression, anxiety, pain, and substance abuse. These symptoms are thought to be caused by neural inflammation combined with a malfunctioning autonomic nervous system following injury, called dysautonomia. Dysautonomia leads to changes in heart and respiratory rate, increased fatigue, and has been shown to be able to predict future behavioral outcomes such as depression. Vagal nerve stimulation (VNS) is currently being examined as a treatment for blast trauma, as the vagus nerve helps regulate the autonomic nervous system. I predict that VNS following blast exposures will reduce the neural inflammation and severity of dysautonomia following blast TBIs, in turn lessening the chronic behavioral dysfunction that normally occurs after blasts. To examine the effects of VNS on TBIs, I looked at both vital signs and behavioral dysfunction immediately and chronically following the blast. A shock tube that generates clinically relevant overpressure waves was utilized to simulate chronic (3x blast exposures (or sham exposure)) in 11-week-old C57BL/6J mice (n=5-6 per group, from two cohorts of mice). In addition to analyzing cytokine expression to determine inflammation and vital signs, several behavioral assays were run, including: operant conditioning, t-maze, y-maze, acoustic startle, and photophobia, to examine both biological and behavioral changes following blast and VNS treatment. Thus far it appears likely that the VNS treatment has lessened the severity of multiple measures of dysfunction, including pain, startle sensitization, and inflammation. This demonstrates that VNS could be a potential therapeutic for blast TBIs but further research, including running more mice to obtain a larger sample size, is necessary to draw more conclusions.

Blast exposure via detonation of explosives is a major source of trauma for service members, Veterans, and civilian bystanders, resulting in mild traumatic brain injury, post-traumatic stress disorder, and chronic pain. The combination of these effects characterizes the polytrauma clinical triad and is a risk factor for increased substance use and substance use disorder (SUD). Exposure to polytrauma can result in disparate symptom trajectories. Research focused on understanding how distinct symptom trajectories map onto substance preference and SUD risk is the focus of my project. In order to understand the interactions between polytrauma and SUD risk, I use a rodent model that utilizes custom, in-house-built polysubstance self-administration chambers to measure water, alcohol, and fentanyl intake. Two types of tracking are used to monitor drinking: Radio Frequency Identification (RFID) tracking and the volumetric drinking monitor. With these two tracking types combined, we can see which mouse drinks what liquid for what period of time. For this project, I used C57Bl/6 male and female mice aged 9 weeks on arrival. These mice were then single or group-housed in the RFID cages for one week to monitor drinking patterns. Previously, I have tested 48 mice, and found that when one (fentanyl or alcohol) substance was available, mice that were single-housed drank more substance than mice that were group housed. To continue this project, I plan to test fentanyl and alcohol in the same cage, which can give us valuable insight into substance preference and polysubstance use, making it more representative of the human experience, as many people consume multiple drugs at the same time. I also plan to look into sex differences and see how that could alter substance use. These factors combined will give valuable insight into classifying substance use that can lead to more optimized treatment for Veterans with polytrauma.

Associations between aggression and social functioning have been identified among typically developing individuals; however, the link between aggression and social concerns among individuals with Autism Spectrum Disorder (ASD) is not well understood. While aggression is not considered a defining feature of ASD, it is observed at elevated levels and is associated with increased risk of harm to the self and others, and reduced opportunities for social relationships and learning. The aims of this study are to determine whether 1) there is a concurrent association between aggression and impairments in social functioning among children with ASD, 2) aggression can predict social functioning among children with ASD six months later (T2), and 3) peer relationships mediate the potential link between aggression and social deficits among children with ASD. We analyzed data from 399 (ASD- 280) children ages 6 -11 years who were enrolled in the Autism Biomarkers Consortium for Clinical Trials (ABC-CT). We used Spearman’s correlation to understand a potential relationship between aggression and social functioning. To investigate whether aggression can be used to predict social impairments, we compared a series of linear models with social ability at T2 as the outcome variable, and age, sex, aggression, and group (ASD or typically developing) as predictor variables. Peer relationship data was only available for participants with ASD so we ran a mediation analysis among this group to determine whether peer relationships mediate the link between aggression and social deficits. We hypothesized that there would be a positive association between aggression and concurrent social impairments, the best-fit model would include aggression as a predictor variable, and peer relationships would mediate the link between aggression and social impairments. The results of this study may provide information that can be used to optimize interventions for individuals with ASD, improving their quality of life.

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by communication and social impairments deficits. I am investigating alpha power, EEG activity between 8 and 12 Hz, which is found present during quietly resting with greater power (amplitude squared) when the individual's eyes are closed rather than when their eyes are open. This is thought to reflect increased cortical activation (decreased power) when eyes are open in preparation for attention and cognition. Previous research shows lower alpha power in individuals with ASD as compared to typically developing (TD) and has been associated with attention and cognition and social understanding and imitation in individuals with ASD. I plan to evaluate the association between eyes-closed EEG resting-state alpha power and social responsiveness in youth with and without ASD by gender and diagnosis. We collected data from 152 youth (ASD = 76) ages 8-17 years from an NIH-funded study investigating sex and gender differences in youth with ASD. The youth sat with their eyes closed while we collected high-density EEG data. A fast fourier transformation (FFT) was applied to artifact-free EEG data to extract alpha power for central and posterior electrodes. I calculated the Regions of interest (ROIs) for alpha power over: left frontal, mid frontal, right frontal, left central, left posterior, mid central, mid posterior, right central, and right posterior. Parents of children completed the Social Responsiveness Scale-2 (SRS-2), a 65-item rating scale measuring autism traits. I conducted a series of one-way ANOVAs, and correlations to evaluate the differences in frontal, central, and posterior alpha power x group (ASD/TD), gender (male/female), and associations with social responsiveness. Preliminary results comparing alpha power and social responsiveness show significant group differences. This research is important since it helps individuals currently affected with ASD and helps understand the larger class of neurodevelopmental disorders.

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by social-communicative impairments and restricted, repetitive behaviors (RRBs). Children with ASD often have impairments in executive functioning (EF), a broad term for cognitive processes including working memory, planning, impulse control, inhibition, and mental flexibility. Previous research has identified direct links between EF problems with RRBs in children with ASD. Intrinsic brain activity measured by Resting-State EEG (RS-EEG), particularly activation of beta (12-30 Hz) and theta (4-8 Hz) bands, has been shown to reflect memory and inhibition. Prior RS-EEG research has found links between reduced theta and beta activation and increased alpha activation in children with ASD. Thus, this project examines the associations between EF problems and resting-state alpha, beta, and theta activation. This study includes 217 participants, ages 8-17 years (ASD=109) from the ACE GENDAAR network, a four-site NIH-funded project investigating sex differences in individuals with autism. ASD diagnosis was confirmed via gold-standard diagnostic measures. EF was assessed using the Behavior Rating Inventory of Executive Function (BRIEF), a parent-report with eight clinical scales measuring specific EF abilities. RS-EEG was measured using an eyes-open paradigm where children sat and looked at screensaver-like videos. Movement and blink artifacts were rejected, and Fast-Fourier Transformation was performed over clean segments. Average power in the alpha, beta, and theta bands was abstracted over 9 different areas across the head for each subject. We hypothesize that EF abilities in participants will correlate positively with theta and beta activity levels and negatively with alpha activity, specifically in the frontal regions. We hypothesize that, similar to children with ADHD, ASD participants will have a greater theta-beta ratio than typically developing (TD) participants. This study’s data can provide a better understanding of the relation between ASD and EF and identify biomarkers to distinguish between ASD and other EF dysfunction prognoses.

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder that often results in deficits in communication, social skills, and emotion regulation. Additional concerns include disruptions to the sleep wake cycle that result from circadian rhythm dysfunction. Individuals with ASD also generally demonstrate difficulty with emotional regulation, including internalizing behaviors that can present as disorders such as anxiety, depression, general mood disturbance, and externalizing behaviors such as aggression. Previous studies have suggested an association between increased sleep dysfunction and increased issues with internalizing/externalizing behaviors in children, and have insinuated a possible bidirectional relationship between the two. This study aimed to look at the relationship between sleep quality and emotion regulation (internalizing and externalizing behaviors) in adults with and without ASD. 80 adults (ASD=42) from the longitudinal Autism Center of Excellence 2 study, a five-site NIH-funded study on gender differences in autism, were included in the analysis. Autism diagnosis was confirmed via standardized measures. Participants completed the Pittsburgh Sleep Quality Index (PSQI), a measure of sleep quality and the adult self report (ASR), a measure of internalizing and externalizing behaviors. Analysis included correlations between sleep quality and internalizing and externalizing scores for ASD and typically developing (TD) groups. We hypothesized that individuals with ASD will demonstrate higher internalizing and externalizing scores and higher rates of sleep issues; and stronger correlation between the two. Additionally we explored gender differences in sleep quality within the sample. I hypothesized that decreased sleep quality and increased sensitivity to stimuli will increase emotional regulation problems in individuals with ASD. This study might provide evidence to support the increased use of therapy or behavioral interventions for better sleep to improve mental health in individuals with autism.

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by social, communicative, cognitive, and behavioral impairments, observable in over 2% of adults in the United States. Hedonic capacity is a concept that is defined as reward responsiveness and has become associated with ASD and related disorders. When faced with stimuli that are typically considered to be rewarding, one’s level of hedonic capacity can be determined by his or her ability to feel pleasure in response. Research has shown that individuals with ASD experience decreased levels of social and interpersonal pleasure when compared to typically developing (TD) individuals. Considering the prevalence of social impairments in autism, I aim to analyze the relationship between hedonic capacity for interpersonal pleasure and autism social traits in ASD and TD adults. 99 adults (ASD = 52) from the longitudinal ACE2 study, a five-site NIH-funded study on gender differences in autism, were included in the analysis. Participants completed the Anticipatory & Consummatory Interpersonal Pleasure Scale (ACIPS), which assesses an individual’s ability to look forward to interactions with other people (anticipatory social pleasure) as well as to experience pleasure for social/interpersonal interactions when they occur. We also utilized the Social Responsiveness Scale (SRS-2) to measure participant social responsiveness, including social motivation. I expect ASD individuals to have a lower ACIPS score and more deficits in social responsiveness. Additionally, I expect that individuals with decreased interpersonal pleasure will have greater social responsiveness impairments and expect this relationship to be stronger in the ASD group. Understanding the relationship between hedonic capacity for interpersonal pleasure and social responsiveness will allow us to further understand how decreased social pleasure may be related to autism traits and symptoms. 

The mesolimbic dopamine system mediates various behavioral functions, including motivation and reward association. Midbrain dopamine neurons of the ventral tegmental area (VTA) send projections to the nucleus accumbens (NAc), forming the mesolimbic pathway. It has been hypothesized that two subregions of the NAc, core and shell, are differentially involved in motivation and reward association. Previous studies identified distinct groups of dopamine neurons with synaptic projections to either the core or shell. The goal of this project is to establish the functional roles of core- and shell-projecting VTA neurons in motivated behavior. More specifically, we investigate whether either subpopulation is necessary or sufficient to reinstate an extinguished goal-directed behavior. The optogenetic approach allows for millisecond-precise gain or loss of function in distinct neural circuits. We use transgenic mice engineered to express Cre-recombinase in either core- or shell-projecting dopamine neurons. Cre-dependent targeting confers cell-type specificity while local intracranial injection of light-sensitive opsins via viral vector provides anatomical specificity. After surgical manipulations, mice undergo a conditioning task with separate acquisition, extinction, and reinstatement phases to measure the rate of food reward retrieval. Operant conditioning paradigms provide the ability to dissect how cues, contexts, and instrumental responses influence goal-directed behavior. During acquisition, mice are trained for six days of lever pressing for food reward paired with an audiovisual cue. In the extinction phase, no reward or cue is delivered. During reinstatement, lever pressing is followed by laser stimulation and cue presentation. We hypothesize that VTA-core-projecting and VTA-shell-projecting neurons differentially regulate cued reinstatement behavior. Our project will provide insight into the functional organization of the mesolimbic pathway and its role in motivated behavior. Collectively, these studies will establish the anatomical basis of dissociable aspects of motivation, extinction, and reinstatement of reward-seeking behaviors, which are implicated in substance abuse and obsessive-compulsive disorders.

Research in recent decades has highlighted the influence of descriptive and injunctive norms on young adult behavior in relation to alcohol and cannabis use. Simultaneous alcohol and cannabis use is when these two substances are used a way that their effects overlap. Rates of simultaneous alcohol and cannabis use are higher among young adults and use is associated with more negative outcomes compared to alcohol or cannabis alone. The relationship between norms and simultaneous use has been less examined, especially among higher-risk young adults (e.g., driving under the influence). Additionally, it is not fully understood why norms influence substance use behaviors for some individuals and not others. Past research has linked factors of impulsivity with higher rates of substance use, but how impulsivity may impact the relationship between norms and simultaneous use is not known. For this research I used data from a larger parent study that sampled high-risk young adults who reported both simultaneous alcohol and cannabis use and riding or driving under the influence in the past (6) months. I used moderated regression analyses to assess impulsivity as a moderator of descriptive and injunctive norms on substance use. I hypothesized that for young adults high in impulsivity the association between norms and substance use would be dampened. These findings may suggest interventions that utilize norms (e.g., normative feedback interventions) may be less effective for individuals who are higher on impulsivity factors and that other intervention approaches should be applied for these young adults.