Recent work has disclosed the critical role played by enamel peptides in sex classification of old skeletal remains. In particular, protein AMELY (amelogenin isoform Y) is present in the enamel dental tissue of male individuals only, while AMELX (isoform X) can be found in both sexes. AMELY can be easily detected by LC-MS/MS in the ion extracted chromatograms of the SM(ox)IRPPY peptide (monoisotopic [M + 2 H]+2 mass = 440.2233 m/z). In this paper, we exploited the dimorphic features of the amelogenin protein to determine the sex of the so-called ‘Lovers of Modena’, two Late Antique individuals whose skeletons were intentionally buried hand-in-hand. Upon discovery, mass media had immediately assumed they were a male-female couple, even if bad preservation of the bones did not allow an effective sex classification. We were able to extract proteins from the dental enamel of both individuals (~1600 years old) and to confidently classify them as males. Results were compared to 14 modern and archaeological control samples, confirming the reliability of the ion chromatogram method for sex determination. Although we currently have no information on the actual relationship between the ‘Lovers of Modena’ (affective? Kin-based?), the discovery of two adult males intentionally buried hand-in-hand may have profound implications for our understanding of funerary practices in Late Antique Italy.

Supplementary Note 1. MRD-VP-1/1: Detailed description of discovery and preservation 1.1. Recovery. MRD was recovered from the surface in two major pieces. The facial component was found on February 10, 2016 by an Afar worker (Ali Bereino) at a place locally known as "Miro Dora" (11º32'59.5" N; 40º27'54.6" E). The rest of the cranium was found on the same day by YHS ca. 3m northeast of the face. After the discovery of the two major pieces (the face and the neurocranium), a sieving operation was carried out to recover more pieces that might have broken away from the specimen. The team first crawled the area in the immediate vicinity of the two major pieces and then surface swept an area of ca. 30 m 2. While the specimen eroded out from a sandstone horizon (based on the matrix adhered to it) and the amount of loose original sediment on top of it was minimal, the main area that had to be sieved was covered by up to 30 cm deep goat droppings piled up for many years, indicating that there was a lot of animal movement in the area and some of the fragments of MRD-VP-1/1 may have been destroyed by trampling. The sieving operation resulted in the recovery of many fragments including two pieces of the left zygomatic bone. The larger piece constituted most of the malar region including the frontal process and some part of the frontal bone at the superolateral corner of the orbit. The other piece was a small fragment that joined to the bigger piece at the anterior origin of the zygomatic arch. One small but critical piece, that joined the left zygomatic bone with the frontal, was also found from the sieve. Other pieces that were found as a result of the sieving operation include a small fragment of the left zygomatic arch, which joined the temporal bone at the posterior end of the arch. Two more fragments of the zygomatic arch from mid-section were also recovered even though their position on the arch is unknown. A larger piece from the base of the right zygomatic was also recovered. This fragment has the entire masseteric origin preserved but did not join the * Landmarks identifiable on the left and right sides. ** Semilandmarks identified on the curves. * Landmarks identifiable on the left and right sides. ** Semilandmarks identified on the curves.

The 4th century BC marks the main entrance of Celtic populations in northern Italy. Their arrival has been suggested based on the presence of Celtic customs in Etruscan mortuary contexts, yet up to now few bioarchaeological data have been examined to support or reject the arrival of these newcomers. Here we use strontium isotopes, non-metric dental traits and funerary patterns to unravel the biocultural structure of the necropolis of Monterenzio Vecchio (Bologna, Italy). Subsamples of our total sample of 38 individuals were analyzed based on different criteria characterizing the following analyses: 1) strontium isotope analysis to investigate migratory patterns and provenance; 2) non-metric dental traits to establish biological relationships between Monterenzio Vecchio, 13 Italian Iron age necropolises and three continental and non-continental Celtic necropolises; 3) grave goods which were statistically explored to detect possible patterns of cultural variability. The strontium isotopes results indicate the presence of local and non-local individuals, with some revealing patterns of mobility. The dental morphology reveals an affinity between Monterenzio Vecchio and Iron Age Italian samples. However, when the Monterenzio Vecchio sample is separated by isotopic results into locals and non-locals, the latter share affinity with the sample of non-continental Celts from Yorkshire (UK). Moreover, systematic analyses demonstrate that ethnic background does not retain measurable impact on the distribution of funerary elements. Our results confirm the migration of Celtic populations in Monterenzio as archaeologically hypothesized on the basis of the grave goods, followed by a high degree of cultural admixture between exogenous and endogenous traits. This contribution shows that combining different methods offers a more comprehensive perspective for the exploration of biocultural processes in past and present populations.

The results are consistent with tool-assisted manipulation to remove necrotic or infected pulp in vivo and the subsequent use of a composite, organic filling. Fredian 5 confirms the practice of dentistry-specifically, a pathology-induced intervention-among Late Pleistocene hunter-gatherers. As such, it appears that fundamental perceptions of biomedical knowledge and practice were in place long before the socioeconomic changes associated with the transition to food production in the Neolithic.

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