High-throughput targeted paleoproteomics sex estimation on medieval Great Moravia individuals using MALDI-CASI-FTICR mass spectrometry

Bray, F.; Pilmann Koterova, A.; Garbe, L.; Haegelin, M.; Bertrand, B.; Agossa, K.; Rolando, C.; Veleminsky, P.; Bruzek, J.; Morvan, M.

doi:10.64898/2026.02.17.706309

bioRxiv · 10.64898/2026.02.17.706309

High-throughput targeted paleoproteomics sex estimation on medieval Great Moravia individuals using MALDI-CASI-FTICR mass spectrometry

Bray, F.·Pilmann Koterova, A.·Garbe, L.·Haegelin, M.·Bertrand, B.·Agossa, K.·Rolando, C.·Veleminsky, P.·Bruzek, J.·Morvan, M.

View original source ↗Export metadata ↓

Abstract

The estimation of the biological sex of archeological remains is crucial information in bioarchaeology and forensic anthropology. In recent years, proteomics based on molecular sexual dimorphism have emerged as a preferred method, particularly because of its minimally-invasive approach to extracting amelogenin X and Y proteins from tooth enamel. However, there is an increasing demand to accelerate this process while facilitating the analysis of large archaeological assemblages. This study presents a novel high-throughput targeted paleoproteomics method for biological sex estimation using MALDI-CASI-FTICR mass spectrometry. This approach combines the strengths of existing methods, including ultra-high resolution, significantly reduced processing times, targeted analysis, and scalability to large archaeological sample sets. The method was initially validated on modern individuals with known sex and subsequently applied to 130 adult and juvenile individuals from medieval Great Moravia (present-day Czech Republic). Biological sex was successfully estimated for all but one of the individuals. The results not only provide a more efficient biological sex estimation but also help to resolve a few errors in sex assessment previously encountered with osteomorphological and tooth morphometric techniques. The implementation of this method significantly improves the accuracy and efficiency of biological sex estimation, offering a powerful tool for anthropological research. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=79 SRC="FIGDIR/small/706309v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@1ede7e6org.highwire.dtl.DTLVardef@13d2f5org.highwire.dtl.DTLVardef@17ee44dorg.highwire.dtl.DTLVardef@1be9dd9_HPS_FORMAT_FIGEXP M_FIG C_FIG

Explore related subjects

evolutionary biology Preprint

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Bray, F., Pilmann Koterova, A., Garbe, L., Haegelin, M., Bertrand, B., Agossa, K., Rolando, C., Veleminsky, P., Bruzek, J., Morvan, M.. 2026-02-18. High-throughput targeted paleoproteomics sex estimation on medieval Great Moravia individuals using MALDI-CASI-FTICR mass spectrometry. https://doi.org/10.64898/2026.02.17.706309

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

RELAX does not reproduce its own estimates at default settings, and its output does not show it

Selection-intensity estimates from RELAX are reported as a point value of K with a likelihood-ratio P. We report that, at default settings and on data of ordinary size, the program does not reproduce its own fits. Of 27 enzyme entries refitted under two optimiser configurations, none reproduced its log-likelihood to within 0.01 units; the median change was 103 units, the largest over 3,400, and four verdicts reversed. Eighty null orthologues reproduced none. A byte-identical command returned a distinct likelihood on every repetition, single-threaded, across three releases, and on alignments simulated under the fitted model, where 3.3 per cent of replicates reproduced. The documented random-number seed never reaches the generator when assigned on the command line, yet reads back as the value supplied. PAML localises the cause: its two-ratio model, without site classes, reproduced its log-likelihood for all 288 genes; its site-class models agreed for 27 to 67 per cent. The instability follows the mixture over sites, not the program. The output does not show it: 46 of 410 fits ended with a negative likelihood-ratio statistic, impossible under convergence, and 123 of 410 report a K re-estimated under a domain restriction rather than the unconstrained maximum. Of 234 published studies using RELAX, none reported a seed. Seeding while holding the thread count at one reproduced sixty of sixty runs on twenty genes under two releases; the seed alone reproduced none of five, and no documentation states the second condition. We recommend that fits be repeated and their dispersion published.

evolutionary biology↗

bioRxiv2026

Sequential accumulation of adaptive alleles forms an inversion supergene in deer mice

Supergenes are clusters of co-inherited loci that affect multiple or complex phenotypes. Despite the growing number of chromosomal inversions identified as supergenes in natural populations, their molecular basis and evolutionary history often remain obscure. Here, we identified two candidate genes, Slc45a2 and Npr3, within a 41-Mb inversion supergene in the deer mouse (Peromyscus maniculatus) that respectively drive darker coats and longer tails - two traits associated with forest adaptation. Mice homozygous for the inversion (inv/inv) exhibit elevated Slc45a2 expression in melanocytes relative to the congenic standard genotype (std/std), disrupting pheomelanin production. In parallel, downregulation of Npr3 in inv/inv mouse growth plates prolongs postnatal growth of caudal vertebrae, resulting in tail elongation. Population-level analyses further implicate that this supergene arose through the subsequent accumulation of the Npr3 allele within the inversion, rather than by capturing all beneficial mutations at its origin.

evolutionary biology↗

bioRxiv2026

Toxin structure shapes palatability in a chemically defended butterfly

The toxicity of chemical defences is well studied, but the potential contribution of compound structure to predator deterrence remains largely unexplored. Whether predation acts more strongly on toxicity or unpalatability remains largely untested, partly because few systems allow toxin structure to vary independently of quantity. Heliconius sara larvae provide such a system: those reared on Passiflora auriculata sequester cyclopentenyl cyanogenic glucosides (CGs), while those reared on P. biflora biosynthesise comparable quantities of aliphatic CGs. Using two invertebrate predators, Camponotus floridanus ants and Hierodula membranacea mantids, we tested whether this structural difference affects palatability independent of toxicity. Mantids rejected larvae with cyclopentenyl CGs more often than larvae with aliphatic CGs, despite no detectable difference in total CG content. This pattern was mirrored in extract-based assays with ants, independently of cyanide release: extracts with cyclopentenyl CGs remained deterrent, while extracts with aliphatic CGs did not differ in deterrence from water. Live larvae, by contrast, elicited similar responses from ants regardless of CG structure. These results show that variation in toxin structure can strongly affect palatability, with some compounds conferring greater protection than others. This demonstrates the importance of chemical structural diversity in the evolution of chemical defences.

evolutionary biology↗