One of the major difficulties in paleontology is the acquisition of fossil data from the 10% of Earth's terrestrial surface that is covered by thick glaciers and ice sheets. Here we reveal that DNA and amino acids from buried organisms can be recovered from the basal sections of deep ice cores and allow reconstructions of past flora and fauna. We show that high altitude southern Greenland, currently lying below more than two kilometers of ice, was once inhabited by a diverse array of conifer trees and insects that may date back more than 450 thousand years. The results provide the first direct evidence in support of a forested southern Greenland and suggest that many deep ice cores may contain genetic records of paleoenvironments in their basal sections.The environmental histories of high latitude regions such as Greenland and Antarctica are poorly understood because much of the fossil evidence is hidden below kilometer thick ice sheets (1-3). Here, we test the idea that the basal sections of deep ice cores can act as archives for ancient biomolecules and show that these molecules can be used to reconstruct significant parts of the past plant and animal life in currently ice covered areas.The samples studied come from the basal impurity rich (silty) ice sections of the 2km long Dye 3 core from south-central Greenland (4), the 3km long GRIP core from the summit of the UKPMC Funders Group Author Manuscript UKPMC Funders Group Author ManuscriptGreenland ice sheet (5), and the Late Holocene John Evans Glacier on Ellesmere Island, Nunavut, northern Canada (Fig. 1A,B). The latter sample was included as a control to test for potential exotic DNA because the glacier has recently overridden a land surface with a known vegetation cover (6). As an additional test for long-distance atmospheric dispersal of DNA, we included five control samples of debris-free Holocene and Pleistocene ice taken just above the basal silty samples from the Dye 3 and GRIP ice cores (Fig. 1B). Finally, our analyses included sediment samples from the Kap København Formation from the northernmost part of Greenland, dated to 2.4 million years before present (Ma BP) (1,2).The silty ice yielded only few pollen grains and no macrofossils (7). However, the Dye 3 and John Evans Glacier silty ice samples showed low levels of amino acid racemization (Fig. 1A, insert), indicating good organic matter preservation (8). Therefore, following previous success with permafrost and cave sediments (9-11), we attempted to amplify ancient DNA from the ice. This was done following strict criteria to secure authenticity (12-14), including covering the surface of the frozen cores with plasmid DNA to control for potential contamination that may have entered the interior of the samples through cracks or during the sampling procedure (7). PCR products of the plasmid DNA were obtained only from extracts of the outer ice scrapings but not from the interior, confirming that sample contamination had not penetrated the cores.We could reproducibly PCR amplify short ampli...

(SPB) or y.y.kim@leeds.ac.uk (YYK). 2Structural biominerals are inorganic/organic composites that exhibit remarkable mechanical properties. However, the structure-property relationships of even the simplest building unitmineral single crystals containing embedded macromolecules -remain poorly understood. Here, by means of a model biomineral made from calcite single crystals containing glycine (0-7 mol%) or aspartic acid (0-4 mol%), we elucidate the origin of the superior hardness of biogenic calcite.We analyzed lattice distortions in these model crystals by using x-ray diffraction and molecular dynamics simulations, and by means of solid-state nuclear magnetic resonance show that the amino acids are incorporated as individual molecules. We also demonstrate that nanoindentation hardness increased with amino acid content, reaching values equivalent to their biogenic counterparts. A dislocation pinning model reveals that the enhanced hardness is determined by the force required to cut covalent bonds in the molecules.3 Biominerals such as bones, teeth and seashells are characterized by properties optimized for their functions. Despite being formed from brittle minerals and flexible polymers, nature demonstrates that it is possible to generate materials with strengths and toughnesses appropriate for structural applications 1 . At one level, the mechanical properties of these hierarchically structured materials are modelled as classical composites consisting of a mineral phase embedded in an organic matrix 2 . However, the single crystal mineral building blocks of biominerals are also composites 3 , containing both aggregates of biomacromolecules as large as 20 nm 4,5 and inorganic impurities 6,7 . While it should be entirely possible to employ this simple biogenic strategy in materials synthesis 8,9 , the strengthening and toughening mechanisms that result from these inclusions are still poorly understood 10,11 . This work addresses this challenge by analyzing hardening mechanisms in a simple model biomineral system: calcite single crystals containing known amounts of amino acids. We report synthetic calcite crystals with hardnesses equivalent to those of their biogenic counterparts, and offer a detailed explanation for the observed hardening.Since plastic deformation in single crystals occurs by the motion of dislocations, hardness is enhanced by features that inhibit dislocation motion. The mechanisms by which guest species may harden ionic single crystals generally fall into two categories. Second phase particles directly block dislocation motion, requiring a dislocation to either cut through (shear) a particle or bypass it by a diffusive process to keep going 12 . Solutes (point defects) do not directly block dislocation motion, but the stress fields of the dislocations interact with those associated with misfitting solutes, retarding dislocation motion 12 . Biominerals, notably calcite, often deform plastically by twinning 11 , but since twins grow by motion of "twinning dislocations" 13 , these concep...

When mollusc shells are analysed conventionally for amino acid geochronology, the entire population of amino acids is included, both inter- and intra-crystalline. This study investigates the utility of removing the amino acids that are most susceptible to environmental effects by isolating the fraction of amino acids encapsulated within mineral crystals of mollusc shells (intra-crystalline fraction). Bleaching, heating and leaching (diffusive loss) experiments were undertaken on modern and fossil Corbicula fluminalis, Margaritifera falcata, Bithynia tentaculata and Valvata piscinalis shells. Exposure of powdered mollusc shells to concentrated NaOCl for 48 h effectively reduced the amino acid content of the four taxa to a residual level, assumed to represent the intra-crystalline fraction. When heated in water at 140 °C for 24 h, only 1% of amino acids were leached from the intra-crystalline fraction of modern shells compared with 40% from whole shell. Free amino acids were more effectively retained in the intra-crystalline fraction, comprising 55% (compared with 18%) of the whole shell after 24 h at 140 °C. For fossil gastropods, the inter-shell variability in D/L values for the intra-crystalline fraction of a single-age population was reduced by 50% compared with conventionally analysed shells. In contrast, analysis of the intra-crystalline fraction of C. fluminalis does not appear to improve the results for this taxon, possibly due to variability in shell ultrastructure. Nonetheless, the intra-crystalline fraction in gastropods approximates a closed system of amino acids and appears to provide a superior subset of amino acids for geochronological applications.

The colonization of Eurasia by early humans is a key event after their spread out of Africa, but the nature, timing and ecological context of the earliest human occupation of northwest Europe is uncertain and has been the subject of intense debate. The southern Caucasus was occupied about 1.8 million years (Myr) ago, whereas human remains from Atapuerca-TD6, Spain (more than 780 kyr ago) and Ceprano, Italy (about 800 kyr ago) show that early Homo had dispersed to the Mediterranean hinterland before the Brunhes-Matuyama magnetic polarity reversal (780 kyr ago). Until now, the earliest uncontested artefacts from northern Europe were much younger, suggesting that humans were unable to colonize northern latitudes until about 500 kyr ago. Here we report flint artefacts from the Cromer Forest-bed Formation at Pakefield (52 degrees N), Suffolk, UK, from an interglacial sequence yielding a diverse range of plant and animal fossils. Event and lithostratigraphy, palaeomagnetism, amino acid geochronology and biostratigraphy indicate that the artefacts date to the early part of the Brunhes Chron (about 700 kyr ago) and thus represent the earliest unequivocal evidence for human presence north of the Alps.

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Marine and ice-core records show that the Earth has experienced a succession of glacials and interglacials during the Quaternary (last ~2.6 million years), although it is often difficult to correlate fragmentary terrestrial records with specific cycles. Aminostratigraphy is a method potentially able to link terrestrial sequences to the marine isotope stages (MIS) of the deep-sea record 1,2 . We have used new methods of extraction and analysis of amino acids, preserved within the calcitic opercula of the freshwater gastropod Bithynia, to provide the most comprehensive dataset for the British Pleistocene based on a single dating technique. A total of 470 opercula from 74 sites spanning the entire Quaternary are ranked in order of relative age based on the extent of protein degradation, using aspartic acid (Asx), glutamic acid (Glx), serine (Ser), alanine (Ala) and valine (Val). This new aminostratigraphy is consistent with the stratigraphical relations of stratotypes, sites with independent geochronology, biostratigraphy and terrace stratigraphy [3][4][5][6] . The method corroborates the existence of four interglacial stages between the Anglian (MIS 12) and the Holocene in the terrestrial succession. It establishes human occupation of Britain in most interglacial stages after MIS 15, but supports the notion of human absence during the Last Interglacial (MIS 5e) 7 . Suspicions that the treeless 'optimum of the Upton Warren interstadial' at Isleworth pre-dates MIS 3 are confirmed. This new aminostratigraphy provides a robust framework against which climatic, biostratigraphical and archaeological models can be tested.Despite the importance of the terrestrial record for climate models, the difficulties of assigning specific sedimentary sequences to individual climate cycles restricts the use of these data in climate modelling. The British Quaternary is exceptional for the number of who used the extent of racemization in the amino acid L-isoleucine (to its diastereomer D-alloisoleucine, yielding an A/I value) in non-marine mollusc shells to build an aminostratigraphy of terrestrial sequences that could be linked to the marine oxygen isotope stratigraphy. Following debate concerning certain correlations, we developed a revised method of extraction and analysis 9 . Shells of freshwater gastropods (Bithynia and Valvata) from many of the original sites 10 have been re-analysed, confirming much of the A/I stratigraphy. However, it emerged that within-site and within-stage variability increases in shells from older sites. This variability probably results from diagenetic alteration of the biomineral carbonate from aragonite to the more thermodynamically stable calcite 10,11 .Our new method has five significant revisions, three of which reduced within-site variability. First, inter-species variation was minimised by analysing only a single genus of freshwater gastropod (Bithynia). Second, variability in amino acid concentration and D/L values was significantly lowered when samples were crushed to ≤ 500 μm and exposed ...

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Considerable debate surrounds the age of the Middle Pleistocene glacial succession in East Anglia following some recent stratigraphical reinterpretations. Resolution of the stratigraphy here is important since it not only concerns the glacial history of the region but also has a bearing on our understanding of the earliest human occupation of north-western Europe. The orthodox consensus that all the tills were emplaced during the Anglian (Marine Isotope Stage (MIS) 12) has recently been challenged by a view assigning each major till to a different glacial stage, before, during and after MIS 12. Between Trimingham and Sidestrand on the north Norfolk coast, datable organic sediments occur immediately below and above the glacial succession. The oldest glacial deposit (Happisburgh Till) directly overlies the 'Sidestrand Unio-bed', here defined as the Sidestrand Hall Member of the Cromer Forest-bed Formation. Dating of these sediments therefore has a bearing on the maximum age of the glacial sequence. This paper reviews the palaeobotany and describes the faunal assemblages recovered from the Sidestrand Unio-bed, which accumulated in a fluvial environment in a fully temperate climate with regional deciduous woodland. There are indications from the ostracods for weakly brackish conditions. Significant differences are apparent between the Sidestrand assemblages and those from West Runton, the type site of the Cromerian Stage. These differences do not result from contrasting facies or taphonomy but reflect warmer palaeotemperatures at Sidestrand and a much younger age. This conclusion is suggested by the higher proportion of thermophiles at Sidestrand and the occurrence of a water vole with unrooted molars (Arvicola) rather than its ancestor Mimomys savini with rooted molars. Amino acid racemisation data also indicate that Sidestrand is significantly younger than West Runton. These data further highlight the stratigraphical complexity of the 'Cromerian Complex' and support the conventional view that the Happisburgh Till was emplaced during the Anglian rather than the recently advanced view that it dates from MIS 16. Moreover, new evidence from the Trimingham lake bed (Sidestrand Cliff Formation) above the youngest glacial outwash sediments (Briton's Lane Formation) indicates that they also accumulated during a Middle Pleistocene interglacial -probably MIS 11. All of this evidence is consistent with a short chronology placing the glacial deposits within MIS 12, rather than invoking multiple episodes of glaciation envisaged in the 'new glacial stratigraphy' during MIS 16, 12, 10 and 6.