Identification of Immunoreactive Material in Mammoth Fossils

The fossil record represents a history of life on this planet. Attempts to obtain molecular information from this record by analysis of nucleic acids found within fossils of extreme age have been unsuccessful or called into question. However, previous studies have demonstrated the long-term persistence of peptides within fossils and have used antibodies to extant proteins to demonstrate antigenic material. In this study we address two questions: Do immunogenic/antigenic materials persist in fossils? and; Can fossil material be used to raise antibodies that will cross-react with extant proteins? We have used material extracted from a well-preserved 100,000-300,000-year-old mammoth skull to produce antisera. The specificity of the antisera was tested by ELISA, western blotting, and immunohistochemistry. It was demonstrated that antisera reacted specifically with the fossils and no the surrounding sediments. Reactivity of antisera with modern proteins and tissues was also demonstrated, as was the ability to detect evolutionary relationships via antibody-antigen interactions. Mass spectrometry demonstrated the response of amino acids and specific peptides within the fossil. Peptides were purified by anion-exchange chromatography and sequenced by tandem mass spectrometry. The collagen-derived peptides may have been the source of at least some of the immunologic reactivity, but the antisera identified molecules that were not observed by mass spectrometry, indicating that immunologic methods may have greater sensitivity. Although the presence of peptides and amino acids was demonstrated, the exact nature of the antigenic material was not fully clarified. This report demonstrated that antibodies may be used to obtain information from the fossil record

The Ursinus Weekly, April 16, 1951

May Day cast chosen; Practices begin this week • Arsenic and Old Lace is underway • Methods discussed by archaeologist • John Billman named head of IRC for coming year • McClure urges students to take army tests • French Club to present pianist, vocalists in recital • 35 Weekly workers honored at banquet • Varsity Club plans big weekend; Variety show, dance listed April 27, 28 • Ursinus to be honored by Newcomen Society May 24 • Self-help jobs available; Letters must be written • Many jobs available to Spring graduates • \u2751ers forced to break up as Fisher must leave for U.S. Navy service • Editorials: Opinions sought; Honor comes to Ursinus • Letters to the editor • Advice to book buyers • Truman decision upheld • Junior Prom traditions seen through years; Theme varies • Poll reveals students\u27 views on MacArthur ouster • Local character, Little Mink, writes parents about college • Preparing catalogue is year-round job, Dr. Phillips says • Moravian beats Ursinus in first home game, 7-3 • Mrs. Whiting stars as player, coach • Bruin team loses fourth straight to Temple nine • Bears defeated in trial meet • Movie to be shown • Two fireside chats planned for separate meetings April 18https://digitalcommons.ursinus.edu/weekly/1565/thumbnail.jp

The Ursinus Weekly, May 7, 1951

Bill LeKernec to edit next year\u27s Lantern • Sororities elect new officers, plan events • Varsity banquet tonight • W. A. A. holds election • Y members plan 1951-52 activities at camp retreat • Campus ready for May Day; Pageant, play to be big events • Professors tackle broad questions in first panel • YM-YW name cabinet, heads of commission • Graduation announcements available to seniors • Students cast primary votes for officers • Election of Curtain Club officers listed • Chi Alpha to elect • Editorials: System a success • Making of foreign policy • Letters to the editor • Three officials embroiled • Ursinus mentioned in Gramercy Ghost • May Day histories reveal variety of festivities • Weekly scribe sheds light on life of famous Ursinus College athlete • Local lassies win 4-1 over Rosemont • Interfraternity track meet to begin this Wednesday • Temple women defeat local tennis squad on May 2 • Cindermen win as Eshbach, Scheirer and Loomis are double winners • Bears capitalize on four hits to beat Garnet • Moravian, Elizabethtown suffer as netmen extend streak to four • Ursinus enters three in intercollege tennis • Bearettes shut-out Albright squad • Lincoln nine halts rally to defeat Grizzlies, 6 to 3 • Curtis clinches first place slots in both leagues • Women\u27s softball team defeats Drexel, Temple • Forum speaker tells of Turkey\u27s position • Dr. Rice invited to Washington for Atlantic Union Conference • Ann Knaur elected President of French Club • Rec center party planned • Women day students hold senior dinner • Supper tickets on sale • Chess Club ends successful season • Shaw named prexyhttps://digitalcommons.ursinus.edu/weekly/1568/thumbnail.jp

Dinosaur peptides suggest mechanisms of protein survival

Eleven collagen peptide sequences recovered from chemical extracts of dinosaur bones were mapped onto molecular models of the vertebrate collagen fibril derived from extant taxa. The dinosaur peptides localized to fibril regions protected by the close packing of collagen molecules, and contained few acidic amino acids. Four peptides mapped to collagen regions crucial for cell-collagen interactions and tissue development. Dinosaur peptides were not represented in more exposed parts of the collagen fibril or regions mediating intermolecular cross-linking. Thus functionally significant regions of collagen fibrils that are physically shielded within the fibril may be preferentially preserved in fossils. These results show empirically that structure-function relationships at the molecular level could contribute to selective preservation in fossilized vertebrate remains across geological time, suggest a ‘preservation motif’, and bolster current concepts linking collagen structure to biological function. This non-random distribution supports the hypothesis that the peptides are produced by the extinct organisms and suggests a chemical mechanism for survival

Empirical Evaluation of Bone Extraction Protocols

The application of high-resolution analytical techniques to characterize ancient bone proteins requires clean, efficient extraction to obtain high quality data. Here, we evaluated many different protocols from the literature on ostrich cortical bone and moa cortical bone to evaluate their yield and relative purity using the identification of antibody-antigen complexes on enzyme-linked immunosorbent assay and gel electrophoresis. Moa bone provided an ancient comparison for the effectiveness of bone extraction protocols tested on ostrich bone. For the immunological part of this study, we focused on collagen I, osteocalcin, and hemoglobin because collagen and osteocalcin are the most abundant proteins in the mineralized extracellular matrix and hemoglobin is common in the vasculature. Most of these procedures demineralize the bone first, and then the remaining organics are chemically extracted. We found that the use of hydrochloric acid, rather than ethylenediaminetetraacetic acid, for demineralization resulted in the cleanest extractions because the acid was easily removed. In contrast, the use of ethylenediaminetetraacetic acid resulted in smearing upon electrophoretic separation, possibly indicating these samples were not as pure. The denaturing agents sodium dodecyl sulfate, urea, and guanidine HCl have been used extensively for the solubilization of proteins in non-biomineralized tissue, but only the latter has been used on bone. We show that all three denaturing agents are effective for extracting bone proteins. One additional method tested uses ammonium bicarbonate as a solubilizing buffer that is more appropriate for post-extraction analyses (e.g., proteomics) by removing the need for desalting. We found that both guanidine HCl and ammonium bicarbonate were effective for extracting many bone proteins, resulting in similar electrophoretic patterns. With the increasing use of proteomics, a new generation of scientists are now interested in the study of proteins from not only extant bone but also from ancient bone

Molecular tests support the viability of rare earth elements as proxies for fossil biomolecule preservation

The rare earth element (REE) composition of a fossil bone reflects its chemical alteration during diagenesis. Consequently, fossils presenting low REE concentrations and/or REE profiles indicative of simple diffusion, signifying minimal alteration, have been proposed as ideal candidates for paleomolecular investigation. We directly tested this prediction by conducting multiple biomolecular assays on a well-preserved fibula of the dinosaur Edmontosaurus from the Cretaceous Hell Creek Formation previously found to exhibit low REE concentrations and steeply-declining REE profiles. Gel electrophoresis identified the presence of organic material in this specimen, and subsequent immunofluorescence and enzyme-linked immunosorbant assays identified preservation of epitopes of the structural protein collagen I. Our results thereby support the utility of REE profiles as proxies for soft tissue and biomolecular preservation in fossil bones. Based on considerations of trace element taphonomy, we also draw predictions as to the biomolecular recovery potential of additional REE profile types exhibited by fossil bones

Constructing living buildings: a review of relevant technologies for a novel application of biohybrid robotics

Biohybrid robotics takes an engineering approach to the expansion and exploitation of biological behaviours for application to automated tasks. Here, we identify the construction of living buildings and infrastructure as a high-potential application domain for biohybrid robotics, and review technological advances relevant to its future development. Construction, civil infrastructure maintenance and building occupancy in the last decades have comprised a major portion of economic production, energy consumption and carbon emissions. Integrating biological organisms into automated construction tasks and permanent building components therefore has high potential for impact. Live materials can provide several advantages over standard synthetic construction materials, including self-repair of damage, increase rather than degradation of structural performance over time, resilience to corrosive environments, support of biodiversity, and mitigation of urban heat islands. Here, we review relevant technologies, which are currently disparate. They span robotics, self-organizing systems, artificial life, construction automation, structural engineering, architecture, bioengineering, biomaterials, and molecular and cellular biology. In these disciplines, developments relevant to biohybrid construction and living buildings are in the early stages, and typically are not exchanged between disciplines. We, therefore, consider this review useful to the future development of biohybrid engineering for this highly interdisciplinary application.publishe

Soft-tissue evidence for homeothermy and crypsis in a Jurassic ichthyosaur

Ichthyosaurs are extinct marine reptiles that display a notable external similarity to modern toothed whales. Here we show that this resemblance is more than skin deep. We apply a multidisciplinary experimental approach to characterize the cellular and molecular composition of integumental tissues in an exceptionally preserved specimen of the Early Jurassic ichthyosaur Stenopterygius. Our analyses recovered still-flexible remnants of the original scaleless skin, which comprises morphologically distinct epidermal and dermal layers. These are underlain by insulating blubber that would have augmented streamlining, buoyancy and homeothermy. Additionally, we identify endogenous proteinaceous and lipid constituents, together with keratinocytes and branched melanophores that contain eumelanin pigment. Distributional variation of melanophores across the body suggests countershading, possibly enhanced by physiological adjustments of colour to enable photoprotection, concealment and/or thermoregulation. Convergence of ichthyosaurs with extant marine amniotes thus extends to the ultrastructural and molecular levels, reflecting the omnipresent constraints of their shared adaptation to pelagic life

The Association of Systemic Microvascular Changes with Lung Function and Lung Density: A Cross-Sectional Study

10.1371/journal.pone.0050224PLoS ONE712