Title: MEMO-ASAP: Mealtime Effects on Maternal Outcomes- Adolescents on the Autism Spectrum Amongst the COVID Pandemic

Authors: Kate Lindig, Deborah Rafferty, Naomi Ekas, and Chrystyna Kouros

Introduction: Mental health of mothers of children on the autism spectrum has been extensively studied (e.g., Sawyer et al., 2009). Prior studies have found connections between the time commitment and pressure experienced by mothers of autistic children and poor mental health outcomes (Liu et al., 2020). Mealtimes for children on the autism spectrum often create stress for mothers (Ausderau & Juarez, 2013). Children on the autism spectrum often experience various difficulties with feeding and mealtime behaviors, such as limited food variety (Curtin et al., 2015), food neophobia (Kuschner et al., 2015), high rates of food refusal and more restricted food repertoire compared to neurotypical children (Bandini et al., 2019), as well as increased disruptive behaviors around mealtimes (Curtin et al., 2015). While little research has examined how feeding behaviors in autistic children impact mothers’ mental health (e.g., Ausderau & Juarez, 2013), it is possible that these behaviors may exacerbate negative mental health outcomes as mothers spending hours attempting to help their children eat with little fruition likely causes feelings of stress and anxiety. Therefore, this study aims to examine how food restrictiveness and disruptive behavior during mealtimes for children on the autism spectrum affects measures of maternal mental health during the COVID-19 pandemic, which has the potential to elevate stress-levels and other negative mental health outcomes for mothers.

Method: Ninety-seven mothers with a child on the autism spectrum served as participants for this study. All mothers participated in October 2020 as a follow-up to a larger study that began prior to the COVID-19 pandemic. Mothers were mostly white, non-Hispanic (77%), college-educated (45%), with a yearly income greater than $40K (90%). Mothers answered surveys about their children’s mealtime behaviors (BAMBI) and their own mental health (IDAS, NIH PROMIS).

Results: Two separate multiple regressions were performed to examine associations between disruptive mealtime behaviors, such as refusing to stay seated or screaming, and maternal dysphoria and stress. Results indicated a significant, positive association between the number of disruptive behaviors a child or adolescent with ASD exhibited during mealtimes and maternal self-reports of dysphoria, b = 1.24 (SE = .60), t = 2.05, p = .04, R2 = .05, while controlling for everything else in the model. Similarly, there was a significant, positive relation between disruptive mealtime behaviors and maternal self-reports of stress, b = 1.66 (SE = .82), t = 2.02, p = .047, R2 = .09. The relations between limited variety of food and the study outcome variables were non-significant, ps ≥ .09.

Discussion: Autistic children’s disruptive mealtime behaviors were associated with poor maternal mental health outcomes, specifically higher depressive symptoms and greater stress. Consequently, finding ways to lower disruptive mealtime behaviors is important, since they have the potential to impact maternal mental health outcomes. Future research should test various mealtime intervention programs to find the most effective ways for parents to minimize their autistic children’s disruptive mealtime behaviors. Additionally, future research should explore the effects of mealtime behaviors on fathers’ mental health outcomes, since they are often present during their autistic children’s mealtimes. Finally, future research should look specifically at how children on the autism spectrum’s mealtime behaviors at the current time compare to those during the first six months of the COVID-19 pandemic, when the present study’s data was collected.

References:
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food choices in children with and without autism spectrum disorder. Journal of Autism and Developmental Disorders, 45(10), 3308–3315.
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0912-3

Human language is an action wherein one plans for, produces, and terminates sound production. Errors in motor planning and production lead to vocal dysfluency. Motor control of respiration is critical for a myoelastic-aerodynamic sound generation mechanism that is used by humans and many other vocalizing animals. Developing our understanding of how the forebrain assumes control of brainstem respiratory circuitry is essential for understanding language initiation, execution, and termination. Songbirds are an animal model for speech production in humans because of the numerous similarities between song learning and production and language acquisition and speech production. Zebra finches sing a learned song that is composed of a motif of 4-7 syllables. The motif is repeated multiple times to form a song bout. Here we explore how a cell-type specific class of neurons control sound termination in zebra finches (Taeniopygia guttata). In songbirds, motor production of song requires activity in premotor and motor cortical analogues HVC (letters used as proper name) and RA (robust nucleus of the arcopallium), respectively. Using an AAV (adeno-associated virus) as a viral vector and Cre-dependent expression of a red-shifted opsin (ChRmine), neurons in premotor cortex that project to motor cortex (HVCRA neurons) produced and inserted soma-targeted, membrane-bound ion channels that allowed for light-dependent manipulation of ongoing cellular activity. Using real-time recording of singing, optical stimulation was timed with the production of individual song syllables. Light stimulation was delivered while birds produced self-initiated and female-directed songs, as well as during quiet respiration. We measured respiratory pressure in birds while they were singing by inserting a small piece of silastic tubing into the anterior thoracic air sac. Singing was reliably disrupted by optical illumination of HVCRA neurons. During the production of song, optogenetic activation of HVCRA neurons resulted in a truncation of the ongoing song respiratory pattern within ~25 ms for female-directed songs and ~32 ms for self-initiated songs. Measures of ongoing respiration during HVCRA stimulation outside of singing had no measurable effects on respiration. These data demonstrate that ongoing song motor patterns can be interrupted via activation of a specific class of neurons, and that the effect is mediated by suppression of ongoing respiratory patterns. However, the suppression of respiration is state-dependent. When birds typically sing faster (female-directed song), the termination occurred faster compared to when they sing more slowly (self-initiated song). Further, outside of song, optical activation of the neurons did not affect ongoing respiration. This suggests that forebrain control of respiration is “permitted” only during song. The underlying neural mechanisms allowing for this state-dependent switch remain to be elucidated. The forebrain control of song termination requires robust activation of HVCRA neurons to suppress brainstem respiratory circuitry. Similar suppression of respiratory circuits may control the termination of human speech.

Introduction: TBRI is an attachment-based model of care for children and youth who have experienced relational trauma. TBRI has been used in a number of service settings, but there is a lack of research on TBRI in clinical and counseling services.   
 
Purpose: The purpose of this pilot study is to evaluate behaviors and trauma symptoms among foster and adopted children whose families received clinical services using the TBRI Counselor’s Manual. 
 
Methods: This study used a one group, pre-post design. Caregivers seeking clinical services for their foster and adopted children (n=14) completed the Child Behavior Checklist (CBCL) and Trauma Symptoms Checklist for Young Children (TSCYC) or Trauma Symptoms Checklist for Children (TSCC) prior to treatment and again after 18 family sessions with a clinician using the TBRI Counselor’s Manual.  
 
Results: Although statistical analyses were limited by the small sample size, descriptive statistics suggest that CBCL and TSCYC/TSCC scores were trending in positive directions. Specifically, mean t scores for most CBCL and TSCYC/TSCC subscales improved, including mean scores for attention problems, aggressive behaviors, and anger. Further, the percentage of participants with scores in the clinical/borderline range dropped for most subscales, including attention problems, aggressive behaviors, anger, arousal, as well as for the composite scales of internalizing problems and externalizing problems.   
 
Discussion: Preliminary findings suggest that TBRI may help improve behavior and trauma symptoms among adopted and foster children whose families participate in TBRI-based clinical services. Limitations include a high rate of attrition and lack of a control group. Further research is needed to establish the effectiveness of the intervention in improving outcomes for adopted and foster children and families.

Introduction: During emerging adulthood, the dynamics of the parent-child relationship change such that parents need to support their child’s independence and autonomy (Padilla-Walker et al., 2019). Engaging in helicopter parenting, where parents excessively monitor their adult children, is associated with greater mental health problems for emerging adults (Schiffrin et al., 2014). Emerging adults who experience greater helicopter parenting also report higher levels of entitlement (Segrin et al., 2012). As emerging adults transition to attending college, perhaps living away from home for the first time, they may experience challenges that negatively impact their mental health. However, there is a lack of research that focuses on how helicopter parenting impacts well-being specifically related to students’ college experience. Thus, the overall goal of the current study was to examine associations between helicopter parenting and emerging adult’s college-related well-being. A second aim was to determine the extent to which psychological and academic entitlement (i.e., the belief that academic success is deserved and the avoidance of personal responsibility) mediated these associations.

Method: Participants included 657 undergraduate students at two private universities in the United States. Participants completed an online survey composed of several measures evaluating the participants’ parents’ helicopter parenting behaviors and the participants’ own academic entitlement, psychological entitlement, and college related well-being. Measures used in the current analyses included the Helicopter Parenting and Autonomy Supportive Behavior Scale (Schiffrin et al., 2014), the academic entitlement scale (Chowning & Campbell, 2009), the Psychological Entitlement Scale (Campbell et al., 2004), and the College Student Subjective Wellbeing Questionnaire (CSSWQ; Renshaw, 2016).

Results: Multiple mediation was used to investigate the extent to which academic entitlement – externalized responsibilities and entitled expectations -- and psychological entitlement mediated the association between helicopter parenting and well-being related to the college experience. Results revealed that helicopter parenting positively predicted each measure of entitlement, ps ≤ .001; but, only academic entitlement – externalized responsibilities significantly predicted college related well-being (b path), b = -5.82, SE = .71, p ≤ .001. The 95% confidence interval of the indirect effect using 5,000 bootstrap reiterations did not include zero [-1.25, -0.47], suggesting a significant indirect effect of helicopter parenting on college related well-being through academic entitlement – externalized responsibilities.

Discussion: Although helicopter parenting was associated with higher levels of each form of entitlement, only academic entitlement served as a mediator of the association between helicopter parenting and college related well-being. Students with higher levels of externalized responsibility generally avoid taking personal responsibility for their academic difficulties, instead blaming failures on others (e.g., their college professors). It is possible that these students were accustomed to having their parents manage academic tasks, particularly when they were living at home. However, now that they are living away from home and attending college, they feel entitled to expect similar treatment from their professors. These findings are important for college administrators and others working with college students to identify students who may be particularly vulnerable for struggling during the transition to college.

How does Prior Knowledge Impact Students’ Study Order Decisions?
Matthew N. Olivares, Addison P. Williams, Addison L. Babineau, Michelle L. Rivers, Sarah K. Tauber, & Michael A. Pelch

Learning complex concepts is essential for student success, but it is often challenging. To improve student concept learning, researchers have identified study strategies that can significantly increase student performance (Samani & Pan, 2021). One strategy that has a profound effect on complex concept learning is study order. Prior research has found that memory performance is better when concepts are studied in an interleaved order (i.e., studying multiple concepts mixed together) compared to a blocked order (i.e., studying one concept multiple times before moving to the next; Brunmair & Richter, 2019). One factor that may impact students’ study order decisions (i.e., the decision to interleave or to block) is prior knowledge. We hypothesized that students with more knowledge about a topic would choose to interleave more during learning than would students with less knowledge. To evaluate this hypothesis, we conducted a two-part study to explore the study order decisions of undergraduate students enrolled in Introductory Geology at TCU (i.e., “Understanding the Earth”). During session one of the study, students learned to classify categories of rocks (i.e., igneous, metamorphic, sedimentary) by either studying the rocks or by completing practice tests. After each learning trial, students could block their study by selecting to study another example from the same rock category (e.g., study multiple igneous rocks in a row); or they could interleave their study by selecting to study an example from a different rock category (e.g., study one igneous rock, then one sedimentary rock). After the first session, students completed activities and lectures in their Introductory Geology course aimed at increasing their knowledge of rock classifications. Then, students completed session two of the study by restudying the rock categories, making study order decisions, and taking a final test on rock classification. We will examine students’ study order decisions at session one (i.e., low prior knowledge) as compared to their study order decisions at session two (i.e., high prior knowledge).

The goal of the present study was to compare the effects of two presentation formats when teaching Icelandic words through computer-assisted instruction: In the high-density constructed response condition, the participant was shown a word in Icelandic and asked to type the Icelandic word in every trial. In the pair-test condition, most trials simply presented the Icelandic word paired with a word in English and typing was required only in intermittent probe trials. Ten undergraduate students will participate in the study. Five Icelandic words are assigned to each condition. The assignment of stimuli to conditions is counterbalanced across participants. Pre and post-tests included translation trials, in which the participants were either presented with an Icelandic word and asked to give its equivalent in English, or vice versa. Preliminary data suggest similar acquisition curves in both conditions.

Cannabidiol (CBD) has been found to have anxiolytic effects on behavior when injected (Blessing et al, 2015). In Experiment 1, we investigated whether we could replicate these findings in rats using acute voluntary oral consumption of non-pharmaceutical grade CBD oil at 60mg/kg in Long-Evans rats, with a control group consuming coconut oil (COC). A test was given two hours after consumption to observe any anxiolytic effect of CBD using an elevated plus maze. It was hypothesized that CBD rats would spend more time in the open arms of the elevated plus maze than the control group; however, results revealed that there was no difference between groups. Experiment 2 utilized Fischer rats as subjects to observe the difference in anxiolytic behaviors of TgF344-AD and wild-type (WT) rats and determine if our previous test apparatus was a sufficient measure of anxiety. Previous research has found that AD rats spent less time in the open arms of an elevated plus maze than WT counterparts (Pentkowski et al., 2018). There was no significant difference between the WT and AD rats; however, there was a significant difference between the Fischer rats (WT and AD) and Long-Evans rats (CBD and COC) with the Long-Evans rats spending more time in the open arms than the Fischer rats. Results will be discussed regarding possible factors for finding similar behavior across rats.

Development of the Opioid-Treatment Linkage Model Resource Guide to Strengthen Parole Officers’ Role in Promoting Linkage to Community Services for Individuals Involved in the Justice System

Brooke Preston, Jennifer Becan, PhD, Jennifer Lux, PhD, Chelsea Wood, MPH, Amanda L. Wiese, MS, Kevin Knight, PhD

As funded by the National Institute on Drug Abuse, the Justice Community Opioid Innovation Network (JCOIN) seeks to improve health outcomes for individuals who are at risk for using opioids upon release from correctional facilities. Specifically, JCOIN aims to increase linkage and receipt of community substance use and medication for opioid use disorder (MOUD) treatment within 18 communities across Texas, New Mexico, and Illinois. This system-level initiative will leverage and facilitate cooperation between community corrections staff and community-based treatment providers by providing training on innovative and best practices and service provider lists. Specifically, the Opioid-Treatment Linkage Model (O-TLM) Resource Guide is being designed to help provide parole officers knowledge of the Behavioral Health Services Cascade, which outlines the process of screening, assessment, and referral to health providers, and the treatment that clients undergo. The O-TLM Resource Guide includes information on evidence-based practices and resources that will aid parole officers in more effectively guiding their clients through each step of the services cascade. Additionally, local community provider lists and maps are being created to increase parole officers’ awareness of screening, assessment, and treatment facilities within their community. The list includes a comprehensive catalogue of providers, the services they offer, and their contact information. These guides will hopefully make it easier for parole officers to more actively link clients to community substance use and MOUD services that match the specific client needs.

Past research has shown that merely thinking about an attitude object can result in self-generated attitude polarization (Tesser, 1978). The current study examined the effects of a specific type of thought—extrapolating traits about proponents and opponents of a social issue—on participants’ post-manipulation attitudes. Participants completed an online survey in which they either extrapolated traits about people who support or oppose legalized abortions, or listed synonyms of experimenter-provided personality traits. Participants who extrapolated reported more positive attitudes towards those who agreed and more negative attitudes toward those who disagreed with the participant’s position on abortion than participants who wrote synonyms. Additionally, extremity of extrapolated traits predicted more positive (negative) post-manipulation attitudes towards those who agreed (disagreed) with the participant’s position on abortion. Our findings extended past research on mere thought by offering new insights into a specific thought strategy that can lead to attitude polarization.

Historically women and minorities have been underrepresented in the STEM field. What about individuals who identify as minority women? Their representation in the STEM field is even less than that of White women or minority men. How do we change this phenomenon and increase diversity in the STEM field? This is the question many leaders face every day when trying to increase diversity in their STEM oriented companies, college programs, and departments. Previous research has shown that enrichment programs that help “bridge the gap” between the majority and minority in the STEM field can be beneficial to minority individuals' pursuing a career in this area. (Brown et al., 2020). One aspect that contributes to the pursuance and retention of minority individuals in the STEM field is mentorship. The goal of this study is to evaluate the impact of mentorship on female minority high school students that wish to pursue a career in the STEM field. This topic is important to examine because it can help contribute to data on how to diversify the STEM field by targeting students in high school who wish to pursue STEM degrees in college. The mentorship program in this study helps prepare students to pursue STEM degrees in college by pairing them with a mentor who is currently a minority female in college pursuing a STEM degree and addressing topics such as college applications, resume building, mental health, time management, and navigating STEM classes in college. A series of pre-, during-, and post- surveys were administered via Qualtrics during the mentor program in the 2021/2022 academic school year. The surveys assessed participants' attitudes and feelings toward college, their knowledge and understanding of essential things needed to complete a college application, and if they were able to foster a meaningful relationship with their mentor.

The purpose of the present study was to extend previous studies in our lab that evaluated the stability of classes of stimuli that were learned through Equivalence-based instruction (EBI) and Complete Instruction (CI) procedures. Sixty undergraduate students received training to establish three stimulus classes with four members in each class. The students were randomly assigned to two groups: EBI – in which they received training for some of the relations – and CI – that targeted all possible relations between the members of each class. After undergoing training and equivalence test (Phase 1), participants received contingency reorganization training (Phase 2). In the reorganization phase, new relations between stimuli were established as correct. Stability was evaluated in an immediate contingency-reversal post-test. Overall, there was no statistical difference between EBI and CI groups. Participants made more errors in trials that tested derived changed relations than in trials that tested derived unchanged relations between stimuli.

Examining Parental Alcohol Use, Gender, and Peer Relationships as Predictors of Substance Use Severity
Vinisha Inaganti, Brooke Preston, Kha Hoai Bao Vu, Addison Williams

In the United States, issues with substance use among adolescents has grown in its prevalence, and past research has shown a continuity in substance use amongst children with substance involved parents. In adolescence, young men tend to report higher rates of alcohol use when compared to young women. As such, we were interested in examining gender as a moderating factor on the relationship between parental alcohol use and substance use severity. Amazon’s MTurk was used to recruit 185 participants with a history of substance use to complete a series of surveys. Participants reported parental alcohol use during childhood and involvement with pro-social peers. Results revealed gender did not moderate the relationship between parental substance use and respondents’ self-reported substance use. However, there were strong correlations among substance use severity, parental alcohol use, and involvement with pro-social peers. Specifically, respondents who were the child of a mother who experienced problems with alcohol were 3.12 time more likely to have a severe SUD. This effect was not observed when paternal alcohol use was examined as a predictor of substance use severity (p = .651). Respondents involved with pro-social peers are less likely to have a severe substance use disorder (p < .001). Together, children whose mothers struggled with alcohol use were more likely to develop a severe substance use disorder. In contrast, having a father who struggled with alcohol use did not affect respondents’ involvement with substance use. Furthermore, having a peer support system reduces subsequent substance use, regardless of parental alcohol use.

Antibiotic resistance has been increasing rapidly; however, the amount of new and effective antibiotics is declining. One area of growing interest is the use of metal nanoparticles because they are relatively easy to make and can be synthesized into different shapes, sizes, and with various chemical properties. In particular, zinc oxide nanoparticles have shown to be effective against various bacterial strains; however, the mechanism that zinc oxide utilizes to exhibit its antimicrobial activity is still unknown. It is also not clear what properties of zinc oxide such as size or proximity to bacterial cells are critical for its antimicrobial activity. In order to gain a better understanding of the mechanism behind zinc oxide’s antimicrobial activity, we tested Staphylococcus aureus with various zinc oxide particles under different conditions. Specifically, we looked at whether particle size, contact with bacterial cells, and media type influenced antimicrobial activity. Our results suggest that particle size does not influence zinc oxide activity, but media type significantly impacts antimicrobial activity. Physical contact, although more effective, is not absolutely required to see inhibition of bacterial growth. Understanding the mechanisms that zinc oxide utilizes may guide design for future particles that will improve their effectiveness.

Alzheimer’s disease (AD) is a progressive neurodegenerative disease that is projected to affect almost 14 million American adults by the year 2050. While the prevalence of this detrimental disease is rapidly increasing in the United States, researchers have established the key pathologies connected to AD, including the development of extracellular, amyloid beta (Aβ) plaques, and intracellular, hyperphosphorylated, neurofibrillary tau tangles. Overall, AD engenders general atrophy of the brain and damage to key brain regions including the cerebral cortex and hippocampus, the main brain region responsible for the neural mechanisms of learning and memory. AD pathologies develop in these regions, which commonly results in neuronal death. The presence of AD pathologies, such as Aβ, activates microglial cells in the brain. Glial cells are the most common brain cells that provide support to neurons. Microglia specifically serve as resident immune cells in the brain, clearing cellular debris, such as dead neurons. Therefore, microglia play a key role in the progression of several neurodegenerative diseases. The activation of microglial cells results in an increased secretion of effector proteins, known as pro-inflammatory cytokines. These are released when inflammatory agents, such as Aβ, are present in the brain. Microglial cells commonly produce pro-inflammatory cytokines, such as TNF-α. Although microglial activation is advantageous at first, continual activation of microglial cells results in a constant inflammatory state. Chronic inflammation can lead to detrimental tissue damage that plays a vital role in neurodegeneration. Another key AD pathology, oxidative stress, is connected to chronic inflammation. Oxidative stress develops when the antioxidant system is unbalanced, resulting in the accumulation of reactive oxygen species (ROS). The presence of inflammatory agents and ROS have the potential to activate microglial cells. Accordingly, our lab utilizes microglial cells to study the harmful effects of inflammation on the brain. Dr. Kayla Green’s lab in the TCU Chemistry Department has successfully created compounds that act as potent antioxidants, L2 and L4. We collaborate with Dr. Green’s lab to research the possible rescue effects of L2 and L4 against inflammation in immortalized, BV2 microglial cells. In our lab’s previous research, we have demonstrated that both L2 and L4 have the capacity to rescue BV2 cells and increase cell survival during oxidative stress. Moreover, the main purpose of the current experiment is to further study the effects of these compounds against key AD pathologies, to understand their therapeutic potential against inflammation in vitro. In the current experiments, we utilized lipopolysaccharide (LPS), an element from the cell wall of gram-negative bacteria, to induce an inflammatory response in BV2 cells. First, we determined several timepoints and concentrations in which LPS treatment successfully induced the secretion of TNF-alpha. Next, we pre-treated cells with the compound, L4, for one hour prior to LPS treatment, to study the possible rescue effects of the drug against pro-inflammatory cytokine production. We are currently determining which concentration of L4 is the most therapeutic against pro-inflammatory cytokine production in BV2 cells.

Mercury (Hg) is found in the environment in excess of historic baselines throughout the globe because of widespread atmospheric emissions of inorganic mercury (IHg) from anthropogenic sources such as coal-fired power plants and artisanal gold mines. In aquatic ecosystems, Ihg deposited from the atmosphere is converted by bacteria to methylmercury (MeHg), a bioavailable neurotoxin that adversely affects the health of vertebrates including humans and wildlife. Because IHg deposition varies across the landscape, it is necessary to monitor MeHg levels in aquatic food webs of individual waterbodies. This is a challenge because there are millions of river miles and lakes in the U.S. Shoreline spiders that feed on MeHg-contaminated emergent aquatic insects have been proposed as sentinel species to monitor MeHg contamination. Sentinel species are species which serve to map the bioavailable fraction of pollution in an ecosystem by retaining the pollutants in their tissue. The objective of this study was to test the hypothesis that shoreline spiders can be used as sentinels to evaluate MeHg contamination of river food webs. Our study focused on the Clear and West forks of the Trinity River. A pilot study in 2016 indicated the two forks have different levels of MeHg contamination. From June to August 2019, we collected over 1000 long-jawed orb weaver spiders (Tetragnathidae) along the shorelines of the two forks of the river. Spiders were preserved in 95% ethanol and sorted by leg length into different size categories. Mercury was analyzed using direct Hg analysis. Concentrations of Hg in spiders increased with spider size and was higher in the Clear Fork than the West Fork. A follow up study confirmed that fish in the Clear Fork had higher concentrations of MeHg than in the West Fork. This is one of the first studies to demonstrate that shoreline spiders can be used as sentinels of MeHg contamination in river ecosystems.

Across North America, bats are being killed in large numbers at wind energy facilities and there is concern that this level of mortality threatens bat populations. Currently three species of migratory tree bats, including two Lasiurus species, comprise ~75% of all known fatalities; however, as wind energy development expands into new areas (e.g., the southwestern U.S.) there is the potential for new species to be impacted. Ongoing work in our labs has indicated that that our current understanding of the distribution of Lasiurus species across North America is limited, at best, and that more species are impacted by wind energy development than previously thought. Accurate knowledge about which species are being impacted where, and to what extent, will greatly improve the implementation of effective mitigation strategies. We obtained 19 bat fecal samples from wild-caught Lasiurus bats from a study being conducted at Texas State University to improve the species-specific effectiveness of an ultrasonic acoustic deterrent (UAD) at deterring bats from approaching operational wind turbines. Based on morphology, these wild-caught bats were identified as eastern red bats (L. borealis), but it is possible that some of the individuals were western red bats (L. blossevillii). We extracted DNA from the bat fecal samples and amplified the COI mitochondrial gene to determine the correct species identification for each sample. The final sequencing reactions are underway, and the results will be available soon. These data will improve the accuracy of the results from the flight cage study at Texas State University and will contribute to improving strategies to reduce bat fatalities at wind energy facilities.

Although wind energy facilities are a growing source of renewable, clean energy, they have been shown to contribute to increasing bat mortalities which could threaten the persistence of bat populations. This study aims to expand what we know about the biology and behavior of bat species impacted by wind energy development. Recent research has indicated that yellow bats (Lasiurus spp) are killed at wind energy facilities in the Rio Grande Valley of south Texas. We have limited understanding of the population biology or movement patterns in these species, so the extent to which wind turbine mortality may impact these bats is currently unknown. As part of ongoing research in our labs, I extracted DNA from 18 tissue samples collected from northern yellow bats (Lasiurus intermedius) at a wind energy facility in Willacy county, Texas in 2015. I amplified a region of the mtDNA, the COI locus, and will compare genetic diversity of these samples to a larger data set from wind energy facilities in nearby Starr and Hidalgo Counties that were studied in 2016 and 2017. Together, these datasets will improve our understanding of Lasiurus intermedius genetic diversity and population structure, and have the potential to provide much needed insights into the potential impacts of wind energy development on bats in southern North America.

Mutations in BReast CAncer 1 protein (BRCA1) play a crucial role in DNA damage control such as double-strand DNA break repair mechanisms. Mutations in BRCA1 increase the chance of disrupted genetic integrity by its contributions to the development of breast cancer. BRCA1 must bind to its partner protein PABL2 (Partner and Localizer to BRCA2) in order to properly carry out its function in the repair mechanism pathway, but its conformation once bound to PALB2 is not clear. In its inactive state, PALB2 is known to remain in an alpha-helical coiled-coil homodimer conformation. Through this observation, we hypothesized that the intrinsically disordered region of BRCA1 on its binding surface will undergo a conformational change into an alpha-helical form. In order to test this hypothesis, we first created a truncated BRCA1, making it 50 amino acids long, then conducted nuclear magnetic resonance (NMR) experiments. Through the NMR experiments, we found that the binding interface of BRCA1 does change its conformation into a helical state, forming a coiled-coil heterodimer upon binding with PALB2.

The proper functioning of the protein PALB2 is vital to preventing tumor formation within breast tissues in individuals. Upon the detection of DNA damage, PALB2 and BRCA1 bind to each other along with BRCA2 to form a DNA repair complex. This complex then repairs DNA double-strand breaks in order to prevent the accumulation of DNA damage that leads to breast cancer. While both BRCA1 and BRCA2 have been extensively studied, a lot of information about the structure and function of PALB2 remains unknown. It is thought that BRCA1 and PALB2 bind via PALB2’s coiled-coil domain; however, how variants of unknown significance (VUS) affect this binding interaction is largely unknown. Further, while some of these VUS have been studied in vivo, cheaper and easier in vitro methods to measure their effect on binding affinity have yet to be formulated. Thus, we hypothesized that isothermal titration calorimetry (ITC) could be used as an in vitro testing method for assessing the effects of VUS within the coiled-coil domain of PALB2 on the binding event between PALB2 and BRCA1. Further, we hypothesized that a decrease in binding between the two proteins as measured by ITC would correlate with a decrease in DNA repair as measured in vivo. We tested the efficacy of this method by creating seven mutations within the coiled-coil domain of PALB2 and measuring the binding event of PALB2 to BRCA1 via ITC. Our results strongly suggest that the binding event is enthalpic in nature and can be adequately measured via ITC as evidenced by the correlation between our in vitro data and previous in vivo data.

Evaluating sex-specific differences in cellular immune function in a small fish model, the fathead minnow

Andrew Mielcuszny
Department of Biology

Advisor: Dr. Marlo Jeffries

Previous studies in the Jeffries lab have shown that male and female fathead minnows differ in their ability to fight and survive bacterial infections. Specifically, males have significantly higher rates of mortality upon infection than females. Despite this, few studies have sought to identify the sex-specific differences in specific immune processes that underlie the observed differences in survival following pathogen infection. The purpose of this study was to examine the sex-specific differences in phagocytic cell activity, a key innate immune response in which immune cells engulf and destroy pathogens. To evaluate phagocytic cell activity, kidney cells were isolated from male and female adult fathead minnows and their ability to phagocytose fluorescently-labeled E. coli was measured. The relative phagocytic cell activity of male and female fathead minnows will be presented in an effort to explain whether differences in phagocytosis contribute to differences in pathogen resistance.

Mercury (Hg) is found in the environment in excess of historic baselines throughout the globe because of widespread atmospheric emissions of inorganic mercury (IHg) from anthropogenic sources such as coal-fired power plants and artisanal gold mines. In aquatic ecosystems, Ihg deposited from the atmosphere is converted by bacteria to methylmercury (MeHg), a bioavailable neurotoxin that adversely affects the health of vertebrates including humans and wildlife. Because IHg deposition varies across the landscape, it is necessary to monitor MeHg levels in aquatic food webs of individual waterbodies. This is a challenge because there are millions of river miles and lakes in the U.S. Shoreline spiders that feed on MeHg-contaminated emergent aquatic insects have been proposed as sentinel species to monitor MeHg contamination. Sentinel species are species which serve to map the bioavailable fraction of pollution in an ecosystem by retaining the pollutants in their tissue. The objective of this study was to test the hypothesis that shoreline spiders can be used as sentinels to evaluate MeHg contamination of river food webs. Our study focused on the Clear and West forks of the Trinity River. A pilot study in 2016 indicated the two forks have different levels of MeHg contamination. From June to August 2019, we collected over 1000 long-jawed orb weaver spiders (Tetragnathidae) along the shorelines of the two forks of the river. Spiders were preserved in 95% ethanol and sorted by leg length into different size categories. Mercury was analyzed using direct Hg analysis. Concentrations of Hg in spiders increased with spider size and was higher in the Clear Fork than the West Fork. A follow up study confirmed that fish in the Clear Fork had higher concentrations of MeHg than in the West Fork. This is one of the first studies to demonstrate that shoreline spiders can be used as sentinels of MeHg contamination in river ecosystems.

In recent years, macrocycles have emerged to be potential drug leads, as they show to have promise for targeting disease pathways, however their synthesis is quite difficult and has yet to be optimized. Utilizing glycine specifically in macrocycle synthesis was the objective, and this was done by stepwise reactions of successfully adding compounds onto glycine to prepare for cyclization. Cyanuric chloride, BOC-hydrazine, and morpholine were successfully added to glycine, as proven with thin layer chromatography and NMR. However, problems that arose came with purifying the compound for cyclization due to solubility issues. Many attempts utilized column chromatography, but there seems to be promise in utilizing an extraction to purify the compound and prepare for cyclization.

Organic dyes with photophysical properties affected by alterations in the properties of the media, including viscosity, temperature, and polarity, are known as environment-sensitive probes. These probes are widely used in various areas of analytical, biological and material sciences. This poster will describe our initial efforts on designing multi-responsive environment-sensitive probes based on squaric acid scaffolds. Specifically, the incorporation of aminoquinoline moieties produced small molecule viscometers, which have the ability to sense polarity variations of organic solvents. Multiplexing abilities, coupled with modular and facile synthesis, distinguishes these probes from other types.

Many drugs today are small molecules and function through a specific binding with their target. This has proved to be efficient, yet the idea of larger macromolecules being used as drugs has grown more popular because of their flexibility. The issue with these larger molecules is that they have been previously difficult to synthesize. The emphasis of the research is to find an efficient way to synthesize macrocycles, reducing purification processes and side products. All reactions are done in solution and column chromatography is used to purify. An important aspect is testing if this cyclization method is possible with all amino acids or if limitations are present based on the backbone of the molecule. Because macrocycles have proved difficult to synthesize in the past, they are overlooked in the field of drug design. However, with this rather basic process it is possible to create new rules associated with drug design and defy what was once believed about macrocycles.

Organic synthesis and research into the activity and uses for macrocycle compounds have increased in recent years. These compounds proved to be an interesting field of research due to their size and ability to orient in different ways depending on the environment. The synthesis of these molecules is done by using a stable foundation molecule, cyanuric chloride, which is subject to substitution. The compound can be built from there using nucleophilic substitution with various nitrogen-based compounds. Then, in the final steps of the synthesis, the compounds dimerize forming the macrocycle. The amino acid nucleophile used to build the molecule is being varied to build many different compounds. The challenge, however, is to find the most efficient route for synthesis. I have successfully managed to synthesize one macrocycle compound using lysine with a Z protecting group as the starting material. Throughout the synthesis there was great difficulty with the compound’s solubility, therefore the starting material was switched to a BOC protected lysine amino acid. This resulted in better solubility throughout the process and yielded another successful macrocycle. These results demonstrate how the synthesis pathway we used to build these macrocyclic dimers is successful, but the process can be variable, based on the properties of the amino acid. It is recognized how the synthesis of these compounds is only the first step and further research into the properties and actions of the compounds is necessary. However, a pure product and efficient synthesis in making the macrocycle is important to properly access its properties. My further research will specifically test the antibiotic properties, if any, the macrocycles possess.