Project description:The identification of sex-specific peptides in human tooth enamel by nanoflow liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS) represents a quantum leap for the study of childhood and social relations more generally. Determining sex-related differences in prehistoric child rearing and mortality has been hampered by the inability to determine the biological sex of juveniles. We conducted nanoLC-MS/MS to identify sex-specific peptides in the dental enamel of a child from a settlement pit of the Early Bronze Age settlement of Schleinbach, Austria (c. 1950–1850 BC). Four perimortal impression fractures on the skull of the 5–6-year-old child indicate an intentional killing, with a co-buried loom weight as possible murder weapon. Proteomic analysis, conducted for the first time on prehistoric teeth in Austria, determined the child’s sex as male. While we cannot conclusively decide whether the child became the victim of conflicts between village groups or was slain by members of his own community, we suggest that contextual evidence points to the latter. A possible trigger of violence were the follow-on effects of an uncontrolled middle ear infection revealed by the osteological analysis. The boy from Schleinbach highlights the potential for further investigation of gender-biased violence, infanticide and child murder based on the recently developed method of proteomic sex identification.
Project description:Paleofeces are an important source of information to study the evolution of dietary habits and human health. The UNESCO World Heritage region of Hallstatt-Dachstein/Salzkammergut is one of Europe’s oldest cultural and industrial landscapes; its underground salt mines dating back at least to the 14th century BC are one of the few archaeological sites where paleofeces are well preserved. The high salt concentrations and the constant annual temperature at around 8°C inside the isolated Hallstatt mines have perfectly preserved organic archaeological artefacts (e.g. paleofeces, clothing, mining tools) that provide unique insights into the daily life of a progressive community in Hallstatt. Here we subjected human paleofeces dated from the Bronze Age to early Modern Times to an in-depth microscopic, metagenomic and proteomic analysis. This allowed us to reconstruct the diet of the former population and gain insights into their ancient gut microbiome composition. Our dietary survey identified bran and glumes of different cereals as one of the most prevalent plant fragments. This highly fibrous, carbohydrate-rich diet was supplemented with proteins from broad beans and occasionally with fruits, nuts, or animal food. Linked to these traditional dietary habits all ancient miners up to the early Modern times have gut microbiome structures akin to modern non-Westernized individuals which may indicate a shift in the gut community composition of modern Westernized populations due to quite recent dietary and lifestyle changes. When we extended our microbial survey to fungi present in the paleofeces, we observed in one of the Iron Age samples a high abundance of Penicillium roqueforti and Saccharomyces cerevisiae DNA. Genome-wide analysis indicates that both fungi were involved in food fermentation and provide the first molecular evidence for blue cheese and beer consumption during Iron Age Europe.
Project description:We describe the first dental proteomic profiles of Iron Age individuals (c2000-1000 years B.P), collected from the site of Long Long Rak rock shelter (LLR) in northwest Thailand. A bias toward the preservation of small, acidic and hydrophobic amino acids is observed. It is evident that the 212 proteins identified (2 peptide, FDR <1%) comprise a palimpsest of alterations that occurred both ante-mortem and post-mortem. Conservation of particular amino acids has contributed to the identification of amelogenin peptides. A novel MRM method for sexing individuals using the amelogenin protein is described, with four teeth indicating male origin. Stable isotope analysis using carbon and oxygen isotopes highlights the strongly C3 based (~80%) diet of the Long Long Rak cemetery people, which probably comprised rice combined with protein from freshwater fish among other food items. The combination of pathway and isotopic analysis adds weight to the relatively simple C3 based diets having contributed to the enrichment of pathways associated with metabolic conditions and shows capacity for harboring these conditions prior to death. The combination of proteomics and stable isotope analysis provides a complementary strategy for assessing the demography, diet, lifestyle and possible diseases experienced by ancient populations.
Project description:Human skeletal tissue contains an abundance of proteins some of which may be preserved over geological timescales. The profiling of proteins from ancient individuals — or palaeoproteomics —has begun to provide new information about the diseases suffered in past societies. We describe here the first dental palaeoproteomic profiles of Iron Age individuals, collected from the site of Long Long Rak rockshelter in northwest Thailand. We recovered amino acid sequences for thousands of proteins preserved in their dental tissue, however, it is evident that these palaeoproteomic profiles comprise a palimpsest of modifications that occurred both ante-mortem and post-mortem. Palaeoproteomic profiles are able to categorise disease and show the capacity of these individuals for harboring a variety of illnesses prior to death. Here we apply for the first time palaeoproteomic analysis to five prehistoric human teeth from Southeast Asia. We combine this method with stable isotope analysis using δ18O and δ13C values to broadly identify the diet of these individuals. The specimens were collected from log coffins contained within the Iron Age site of Long Long Rak (LLR) rockshelter in Pang Mapha district, Mae Hong Son Province, northwest Thailand.. Radiocarbon dating shows these log coffins to date within the range of 1,960±30 cal. yr BP to 1,636±44 cal. yr BP.
Project description:Disruption of local iron homeostasis is a common feature of neurodegenerative diseases. We focused on dopaminergic neurons, asking how iron transport proteins modulate iron homeostasis in vivo. Inactivation of the transmembrane iron exporter ferroportin had no apparent consequences. However, loss of the transferrin receptor 1, involved in iron uptake, caused profound, age-progressive neurodegeneration with features similar to Parkinson’s disease. There was gradual loss of dopaminergic projections in the striatum with subsequent death of dopaminergic neurons in the substantia nigra. After depletion of 30% of the neurons the mice developed neurobehavioral parkinsonism, with evidence of mitochondrial dysfunction and impaired mitochondrial autophagy. Molecular analysis revealed strong signatures indicative of attempted axonal regeneration, a metabolic switch to glycolysis and the unfolded protein response. We speculate that cellular iron deficiency may contribute to neurodegeneration in human patients Using Ribotag technology, from mouse ventral midbrain lysates, we isolated actively translated mRNA species from control and Transferrin receptor 1-null dopaminergic neurons. Two mouse ages were used 3 wks (early neurodegeration) 10 wks (late neurodegeneration)