Fungi and bacteria occupy distinct spatial niches within carious dentin.

The role of bacteria in the etiology of dental caries is long established, while the role of fungi has only recently gained more attention. The microbial invasion of dentin in advanced caries especially merits additional research. We evaluated the fungal and bacterial community composition and spatial distribution within carious dentin. Amplicon 16S rRNA gene sequencing together with quantitative PCR was used to profile bacterial and fungal species in caries-free children (n = 43) and 4 stages of caries progression from children with severe early childhood caries (n = 32). Additionally, healthy (n = 10) and carious (n = 10) primary teeth were decalcified, sectioned, and stained with Grocott's methenamine silver, periodic acid Schiff (PAS) and calcofluor white (CW) for fungi. Immunolocalization was also performed using antibodies against fungal ß-D-glucan, gram-positive bacterial lipoteichoic acid, gram-negative endotoxin, Streptococcus mutans, and Candida albicans. We also performed field emission scanning electron microscopy (FESEM) to visualize fungi and bacteria within carious dentinal tubules. Bacterial communities observed included a high abundance of S. mutans and the Veillonella parvula group, as expected. There was a higher ratio of fungi to bacteria in dentin-involved lesions compared to less severe lesions with frequent preponderance of C. albicans, C. dubliniensis, and in one case C. tropicalis. Grocott's silver, PAS, CW and immunohistochemistry (IHC) demonstrated the presence of fungi within carious dentinal tubules. Multiplex IHC revealed that fungi, gram-negative, and gram-positive bacteria primarily occupied separate dentinal tubules, with rare instances of colocalization. Similar findings were observed with multiplex immunofluorescence using anti-S. mutans and anti-C. albicans antibodies. Electron microscopy showed monomorphic bacterial and fungal biofilms within distinct dentin tubules. We demonstrate a previously unrecognized phenomenon in which fungi and bacteria occupy distinct spatial niches within carious dentin and seldom co-colonize. The potential significance of this phenomenon in caries progression warrants further exploration.

Describing Physical Activity Patterns of Truck Drivers Using Actigraphy.

Background: Truck driving is a highly sedentary occupation that places workers at risk for chronic health conditions, such as obesity and high blood pressure. The primary purpose of this study was to objectively describe truck drivers' typical physical activity (PA) patterns. Methods: We used ∼7-10-day baseline PA actigraphy data samples from drivers in the Safety & Health Involvement For Truckers (SHIFT) study (n = 394). Driver PA patterns (e.g., average number of ≥10 minute Freedson bouts per week, time in bouts, and common days/times for PA) were summarized with descriptive analyses. We also compared objective accelerometer data to self-reports. Results: Drivers' weekly PA averaged 14.4 minutes (SD = 37.0), and most PA occurred between 5-6 pm on Tuesdays and Wednesdays. Drivers overestimated self-reported weekly exercise by over 60 min/week compared to accelerometer data. Conclusion: Our results suggest that objective PA assessment may be warranted over self-report when possible, and timing may be key in future PA intervention work with truck drivers.