Development of gene expression-based biomarkers of exposure to metals and pesticides in the freshwater amphipod Hyalella azteca

Ecological risk assessment (ERA) is a framework for monitoring risks of exposure and adverse effects of environmental stressors to populations or communities of interest. One tool of ERA is the biomarker, which is a characteristic of an organism that reliably indicates exposure to or effects of a stressor like chemical pollution. Traditional biomarkers which rely on characteristics at the tissue level and higher often detect only acute exposures to stressors. Sensitive molecular biomarkers may detect lower stressor levels than traditional biomarkers, which helps inform risk mitigation and restoration efforts before populations and communities are irreversibly affected. In this study I developed gene expression-based molecular biomarkers of exposure to metals and insecticides in the model toxicological freshwater amphipod Hyalella azteca. My goals were to not only create sensitive molecular biomarkers for these chemicals, but also to show the utility and versatility of H. azteca in molecular studies for toxicology and risk assessment. I sequenced and assembled the H. azteca transcriptome to identify reference and stress-response gene transcripts suitable for expression monitoring. I exposed H. azteca to sub-lethal concentrations of metals (cadmium and copper) and insecticides (DDT, permethrin, and imidacloprid). Reference genes used to create normalization factors were determined for each exposure using the programs BestKeeper, GeNorm, and NormFinder. Both metals increased expression of a nuclear transcription factor (Cnc), an ABC transporter (Mrp4), and a heat shock protein (Hsp90), giving evidence of general metal exposure signature. Cadmium uniquely increased expression of a DNA repair protein (Rad51) and increased Mrp4 expression more than copper (7-fold increase compared to 2-fold increase). Together these may be unique biomarkers distinguishing cadmium and copper exposures. DDT increased expression of Hsp90, Mrp4, and the immune response gene Lgbp. Permethrin increased expression of a cytochrome P450 (Cyp2j2) and decreased expression of the immune response gene Lectin-1. Imidacloprid did not affect gene expression. Unique biomarkers were seen for DDT and permethrin, but the genes studied were not sensitive enough to detect imidacloprid at the levels used here. I demonstrated that gene expression in H. azteca detects specific chemical exposures at sub-lethal concentrations, making expression monitoring using this amphipod a useful and sensitive biomarker for risk assessment of chemical exposure.

SELECTED CHARACTER PIECES OF FRÉDÉRIC CHOPIN (1810-1849), FRANZ LISZT (1811-1886) AND JOHANNES BRAHMS (1833-1897)

During the 19th century, Frédéric Chopin (1810-1849), Franz Liszt (1811- 1886), and Johannes Brahms (1833-1897) were among the most recognized composers of character pieces. Their compositions have been considered a significant milestone in piano literature. Frédéric Chopin (1810-1849) did not give descriptive titles to his character pieces. He grouped them into several genres such as Mazurkas, Polonaises. His Mazurkas and Polonaises are influenced by Polish dance music and inspired by the polish national idiom. Franz Liszt (1811-1886) was influenced in many ways by Chopin, and adopted Chopin’s lyricism, melodic style, and tempo rubato. However, Liszt frequently drew on non-musical subjects (e.g., art, literature) for inspiration. “Harmonies poétiques et religieuses” and “Années de pèlerinage” are especially representative of character pieces in which poetic and pictorial imagination are reflected. Johannes Brahms (1833-1897) was a conservative traditionalist, synthesizing Romantic expression and Classical tradition remarkably well. Like Chopin, Brahms avoided using programmatic titles for his works. The titles of Brahms’ short character pieces are often taken from traditional lyrical or dramatic genres such as ballade, rhapsody and scherzo. Because of his conservatism, Brahms was considered the main rival of Liszt in the Romantic Period. Brahms character pieces in his third period (e.g., Scherzo Op.4, Ballades of Op.10, and Rhapsodies of Op.79) are concise and focused. The form of Brahms’ character pieces is mostly simple ternary (ABA), and his style is introspective and lyrical. Through this recording project, I was able to get a better understanding of the styles of Chopin, Brahms and Liszt through their character pieces. This recording dissertation consists of two CDs recorded in the Dekelboum Concert Hall at the University of Maryland, College Park. These recordings are documented on compact disc recordings that are housed within the University of Maryland Library System.

Developmental Neural Correlates of Social Interaction

Children develop in a sea of reciprocal social interaction, but their brain development is predominately studied in non-interactive contexts (e.g., viewing photographs of faces). This dissertation investigated how the developing brain supports social interaction. Specifically, novel paradigms were used to target two facets of social experience—social communication and social motivation—across three studies in children and adults. In Study 1, adults listened to short vignettes—which contained no social information—that they believed to be either prerecorded or presented over an audio-feed by a live social partner. Simply believing that speech was from a live social partner increased activation in the brain’s mentalizing network—a network involved in thinking about others’ thoughts. Study 2 extended this paradigm to middle childhood, a time of increasing social competence and social network complexity, as well as structural and functional social brain development. Results showed that, as in adults, regions of the mentalizing network were engaged by live speech. Taken together, these findings indicate that the mentalizing network may support the processing of interactive communicative cues across development. Given this established importance of social-interactive context, Study 3 examined children’s social motivation when they believed they were engaged in a computer-based chat with a peer. Children initiated interaction via sharing information about their likes and hobbies and received responses from the peer. Compared to a non-social control, in which children chatted with a computer, peer interaction increased activation in mentalizing regions and reward circuitry. Further, within mentalizing regions, responsivity to the peer increased with age. Thus, across all three studies, social cognitive regions associated with mentalizing supported real-time social interaction. In contrast, the specific social context appeared to influence both reward circuitry involvement and age-related changes in neural activity. Future studies should continue to examine how the brain supports interaction across varied real-world social contexts. In addition to illuminating typical development, understanding the neural bases of interaction will offer insight into social disabilities such as autism, where social difficulties are often most acute in interactive situations. Ultimately, to best capture human experience, social neuroscience ought to be embedded in the social world.