Chronic non-bacterial osteomyelitis (CNO) is an autoinflammatory disease that predominantly affects children. Osteoclasts are cells that carry out normal bone degradation, and their hyperactivity contributes to pathogenesis of CNO. We do not yet understand how the environment and precursor cells contribute to hyperactivity. We are examining the effects of serum on osteoclastogenesis, the differentiation of osteoclast precursors, from peripheral blood mononuclear cells (PBMCs). Serum from patients with active CNO, inactive CNO, juvenile idiopathic arthritis (JIA), and healthy control participants was collected and used at different concentrations (0.1%, 1%, 10%) to determine their respective effects on osteoclastogenesis. With IRB approval, we sorted PBMCs from ZenBio for monocytes and plated them with media, receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony stimulating factor (M-CSF). Cells were then incubated and media exchanged twice. Tartrate-resistant alkaline phosphatase (TRAP) was used to stain osteoclasts and DAPI was used to stain nuclei. All TRAP positive osteoclasts with three or more nuclei were counted using ImageJ. We then compared counts across serum types and concentrations to identify trends in osteoclastogenesis. Over the course of this project, we encountered issues with the TRAP stain quality and imaging. In response, we modified the procedure at various steps to normalize staining quality across all plates to facilitate the quantification of osteoclast formation. Our results thus far suggest there is a trend of increase in osteoclasts as serum concentration increases with the exception of inactive CNO serum. Given that this study is ongoing and methods are frequently improved, we expect to gain a more comprehensive view of the trends as we progress. The goal of this study is to gain a better understanding of the pathogenesis of CNO, and its results have the potential to guide treatment of CNO and future research.

Music, both vocal and instrumental, captures the attention of children and facilitates language learning. Infants have been shown to prefer infant-directed song over infant-directed speech. Furthermore, education professionals and researchers agree that music supports language acquisition. In early childhood, children are exposed to a great deal of auditory input, including music, in a variety of settings. Much of this exposure occurs in the home, which presents difficulties when attempting to capture data that is truly representative of a child's auditory environment. Thus, the main aim of Tots and Tunes is to quantify and describe the music and speech that infants hear in their home environment during first two years of life. To achieve this, I have randomly extracted 12,000 10-second audio segments from a longitudinal dataset of audio recordings taken of children at ages 6, 10, 14, 18, and 24 months of age by the Language Environment Analysis (LENA) system. This system uses a small device worn by a child throughout their day to record their naturalistic linguistic environment. My team is using a novel citizen-science approach to outsource the annotation of the segments by naive listeners via a platform called Zooniverse. For each segment, we ask participants to listen and subsequently identify whether there is speech and/or music present, whether the speech and/or music is directed to the child wearing the LENA device, and if the speech and/or music is in-person, through an electronic device, or both. In the span of a month, 20% of the segments recorded at 6 months of age have been annotated. I anticipate that our findings will inform parents, researchers, and early childhood professionals about the presence of music in the auditory environment of young children, which will inform further work regarding how best to use music to support language acquisition.