Paget’s disease of bone (PDB) is a chronic skeletal disorder that can affect one or several bones in individuals older than 55 y of age. PDB-like changes have been reported in archaeological remains as old as Roman, although accurate diagnosis and natural history of the disease is lacking. Six skeletons from a collection of 130 excavated at Norton Priory in the North West of England, which dates to medieval times, show atypical and extensive pathological changes resembling contemporary PDB affecting as many as 75% of individual skeletons. Disease prevalence in the remaining collection is high, at least 16% of adults, with age at death estimations as low as 35 y. Despite these atypical features, paleoproteomic analysis identified sequestosome 1 (SQSTM1) or p62, a protein central to the pathological milieu of PDB, as one of the few noncollagenous human sequences preserved in skeletal samples. Targeted proteomic analysis detected >60% of the ancient p62 primary sequence, with Western blotting indicating p62 abnormalities, including in dentition. Direct sequencing of ancient DNA excluded contemporary PDB-associated SQSTM1 mutations. Our observations indicate that the ancient p62 protein is likely modified within its C-terminal ubiquitin-associated domain. Ancient miRNAs were remarkably preserved in an osteosarcoma from a skeleton with extensive disease, with miR-16 expression consistent with that reported in contemporary PDB-associated bone tumors. Our work displays the use of proteomics to inform diagnosis of ancient diseases such as atypical PDB, which has unusual features presumably potentiated by yet-unidentified environmental or genetic factors.

Second-harmonic generation imaging (SHG) captures triple helical collagen molecules near tissue surfaces. Biomedical research routinely utilizes various imaging software packages to quantify SHG signals for collagen content and distribution estimates in modern tissue samples including bone. For the first time using SHG, samples of modern, medieval, and ice age bones were imaged to test the applicability of SHG to ancient bone from a variety of ages, settings, and taxa. Four independent techniques including Raman spectroscopy, FTIR spectroscopy, radiocarbon dating protocols, and mass spectrometry-based protein sequencing, confirm the presence of protein, consistent with the hypothesis that SHG imaging detects ancient bone collagen. These results suggest that future studies have the potential to use SHG imaging to provide new insights into the composition of ancient bone, to characterize ancient bone disorders, to investigate collagen preservation within and between various taxa, and to monitor collagen decay regimes in different depositional environments.

The aim of this study was to investigate the relationship between urbanization and upper respiratory health in two early modern populations from the Netherlands. For this purpose, we analyzed the prevalence of chronic maxillary sinusitis in the adult urban population of Arnhem (n = 83) and in the rural village of Middenbeemster (n = 74). A slightly higher prevalence of chronic maxillary sinusitis was observed in the Arnhem sample (55.4%) compared with the Middenbeemster sample (51.3%), and these variations were not statistically significantly different. Although historical sources attest to the fact that life in the postmedieval settlements of Arnhem and Middenbeemster greatly differed, our results suggest that both environments exposed people to certain respiratory hazards. Furthermore, sinusitis prevalence was also investigated in correlation to sex, as urbanization in the Netherlands often involved women in factory work in direct contrast to the traditional domestic role they kept covering in rural environments. No significant differences were observed between males and females, both in an intersite (Arnhem males vs. Middenbeemster males; Arnhem females vs. Middenbeemster females) and in an intrasite (males vs. females at Arnhem; males vs. females at Middenbemster) comparison. As men and women in Arnhem worked on similar tasks, our results confirm that they were both exposed to similar risk factors. In Middenbeemster, where women mainly stayed inside taking care of the house while men worked the fields, the adverse weather conditions and continuous exposure to pollens and allergens may have enhanced men's chances of contracting chronic maxillary sinusitis. This study suggests that urbanization in the early modern Netherlands was in fact not inherently more detrimental than rural living. Future research incorporating a larger sample from other Dutch sites is being considered to better frame the complex etiology of sinusitis within the present understanding of historic regional variation in urbanization patterns.