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      Proteomic evidence of dietary sources in ancient dental calculus

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          Abstract

          Archaeological dental calculus has emerged as a rich source of ancient biomolecules, including proteins. Previous analyses of proteins extracted from ancient dental calculus revealed the presence of the dietary milk protein β-lactoglobulin, providing direct evidence of dairy consumption in the archaeological record. However, the potential for calculus to preserve other food-related proteins has not yet been systematically explored. Here we analyse shotgun metaproteomic data from 100 archaeological dental calculus samples ranging from the Iron Age to the post-medieval period (eighth century BC to nineteenth century AD) in England, as well as 14 dental calculus samples from contemporary dental patients and recently deceased individuals, to characterize the range and extent of dietary proteins preserved in dental calculus. In addition to milk proteins, we detect proteomic evidence of foodstuffs such as cereals and plant products, as well as the digestive enzyme salivary amylase. We discuss the importance of optimized protein extraction methods, data analysis approaches and authentication strategies in the identification of dietary proteins from archaeological dental calculus. This study demonstrates that proteomic approaches can robustly identify foodstuffs in the archaeological record that are typically under-represented due to their poor macroscopic preservation.

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          The Proteomics Identifications (PRIDE) database and associated tools: status in 2013

          The PRoteomics IDEntifications (PRIDE, http://www.ebi.ac.uk/pride) database at the European Bioinformatics Institute is one of the most prominent data repositories of mass spectrometry (MS)-based proteomics data. Here, we summarize recent developments in the PRIDE database and related tools. First, we provide up-to-date statistics in data content, splitting the figures by groups of organisms and species, including peptide and protein identifications, and post-translational modifications. We then describe the tools that are part of the PRIDE submission pipeline, especially the recently developed PRIDE Converter 2 (new submission tool) and PRIDE Inspector (visualization and analysis tool). We also give an update about the integration of PRIDE with other MS proteomics resources in the context of the ProteomeXchange consortium. Finally, we briefly review the quality control efforts that are ongoing at present and outline our future plans.
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            Diet and the evolution of human amylase gene copy number variation.

            Starch consumption is a prominent characteristic of agricultural societies and hunter-gatherers in arid environments. In contrast, rainforest and circum-arctic hunter-gatherers and some pastoralists consume much less starch. This behavioral variation raises the possibility that different selective pressures have acted on amylase, the enzyme responsible for starch hydrolysis. We found that copy number of the salivary amylase gene (AMY1) is correlated positively with salivary amylase protein level and that individuals from populations with high-starch diets have, on average, more AMY1 copies than those with traditionally low-starch diets. Comparisons with other loci in a subset of these populations suggest that the extent of AMY1 copy number differentiation is highly unusual. This example of positive selection on a copy number-variable gene is, to our knowledge, one of the first discovered in the human genome. Higher AMY1 copy numbers and protein levels probably improve the digestion of starchy foods and may buffer against the fitness-reducing effects of intestinal disease.
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              Microfossils in calculus demonstrate consumption of plants and cooked foods in Neanderthal diets (Shanidar III, Iraq; Spy I and II, Belgium).

              The nature and causes of the disappearance of Neanderthals and their apparent replacement by modern humans are subjects of considerable debate. Many researchers have proposed biologically or technologically mediated dietary differences between the two groups as one of the fundamental causes of Neanderthal disappearance. Some scenarios have focused on the apparent lack of plant foods in Neanderthal diets. Here we report direct evidence for Neanderthal consumption of a variety of plant foods, in the form of phytoliths and starch grains recovered from dental calculus of Neanderthal skeletons from Shanidar Cave, Iraq, and Spy Cave, Belgium. Some of the plants are typical of recent modern human diets, including date palms (Phoenix spp.), legumes, and grass seeds (Triticeae), whereas others are known to be edible but are not heavily used today. Many of the grass seed starches showed damage that is a distinctive marker of cooking. Our results indicate that in both warm eastern Mediterranean and cold northwestern European climates, and across their latitudinal range, Neanderthals made use of the diverse plant foods available in their local environment and transformed them into more easily digestible foodstuffs in part through cooking them, suggesting an overall sophistication in Neanderthal dietary regimes.
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                Author and article information

                Journal
                Proc Biol Sci
                Proc. Biol. Sci
                RSPB
                royprsb
                Proceedings of the Royal Society B: Biological Sciences
                The Royal Society
                0962-8452
                1471-2954
                25 July 2018
                18 July 2018
                18 July 2018
                : 285
                : 1883
                : 20180977
                Affiliations
                [1 ]Department of Archaeology, Max Planck Institute for the Science of Human History , Jena, Germany
                [2 ]Department of Archaeogenetics, Max Planck Institute for the Science of Human History , Jena, Germany
                [3 ]BioArCh, Department of Archaeology, University of York , York, UK
                [4 ]Laboratories of Molecular Anthropology and Microbiome Research, Department of Anthropology, University of Oklahoma , Norman, USA
                [5 ]Institute for Evolutionary Medicine, ETH-Zürich, University of Zürich , Zürich, Switzerland
                [6 ]Functional Genomics Center, ETH-Zürich, University of Zürich , Zürich, Switzerland
                [7 ]Department of Periodontology, College of Dentistry, University of Oklahoma Health Sciences Center , Oklahoma, OK, USA
                [8 ]EvoGenomics, Natural History Museum of Denmark, University of Copenhagen , Copenhagen, Denmark
                [9 ]Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen , Copenhagen, Denmark
                [10 ]Discovery Proteomics Facility, Target Discovery Institute, University of Oxford , Oxford, UK
                [11 ]The Palaeogenomics and Bio-Archaeology Research Network, Research Laboratory for Archaeology and the History of Art, University of Oxford , Oxford, UK
                [12 ]York Osteoarchaeology Ltd , Bishop Wilton, York, UK
                [13 ]Pinellas Dental Specialties , Largo, FL 33776, USA
                [14 ]Department of Anthropology, College of Arts and Sciences, University of Tennessee , Knoxville, TN, USA
                [15 ]Key Laboratory of Vertebrate Evolution and Human Origins of Chinese Academy of Sciences, Institute of Vertebrate Paleontology and Paleoanthropology, Chinese Academy of Sciences , Beijing, People's Republic of China
                [16 ]Department of Anthropology, University of British Columbia , Vancouver, BC, Canada
                Author notes

                Electronic supplementary material is available online at https://dx.doi.org/10.6084/m9.figshare.c.4150424.

                Author information
                http://orcid.org/0000-0002-3718-1058
                http://orcid.org/0000-0002-4528-5877
                http://orcid.org/0000-0003-4226-5501
                http://orcid.org/0000-0002-8991-2318
                http://orcid.org/0000-0002-9715-5951
                http://orcid.org/0000-0002-6808-3370
                http://orcid.org/0000-0002-4183-7574
                http://orcid.org/0000-0002-4092-0392
                http://orcid.org/0000-0003-0763-7592
                http://orcid.org/0000-0001-7128-9903
                Article
                rspb20180977
                10.1098/rspb.2018.0977
                6083251
                30051838
                bbb18d34-51de-4934-b09e-b22997069181
                © 2018 The Authors.

                Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

                History
                : 10 May 2018
                : 25 June 2018
                Funding
                Funded by: Wellcome Trust, http://dx.doi.org/10.13039/100004440;
                Award ID: 108375/Z/15/Z
                Funded by: Kennedy Trust Fund;
                Funded by: White Rose University Consortium Collaboration Fund;
                Funded by: The Max Planck Society;
                Funded by: National Science Foundation, http://dx.doi.org/10.13039/100000001;
                Award ID: BCS-1516633
                Award ID: BCS-1523264
                Award ID: BCS-1643318
                Funded by: DNRF Niels Bohr Professorship;
                Award ID: DNRF128
                Funded by: University of York C2D2 Research Priming Fund (Wellcome Trust);
                Award ID: 097829/Z/11/A
                Funded by: Oxford University Fell Fund;
                Award ID: EBD-10130
                Categories
                1001
                144
                129
                Palaeobiology
                Research Article
                Custom metadata
                July 25, 2018

                Life sciences
                proteomics,dental calculus,dietary reconstruction,mass spectrometry
                Life sciences
                proteomics, dental calculus, dietary reconstruction, mass spectrometry

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