Amino Acids In Bone Collagen Research Articles

Tissue-specific carbon isotope patterns of amino acids in southern sea otters.

The measurement of stable isotope values of individual compounds, such as amino acids (AAs), has become a powerful tool in animal ecology and ecophysiology. As with any emerging technique, questions remain regarding thecapabilities and limitations of this approach, including how metabolism and tissue synthesis impact the isotopic values of individual AAs and subsequent multivariate patterns. We measured carbon isotope (δ13C) values of essential (AAESS) and nonessential (AANESS) AAs in bone collagen, whisker, muscle, and liver from ten southern sea otters (Enhydra lutris nereis) thatstranded in Monterey Bay, California. Sea otters in this population exhibit high degrees of individual dietary specialization, making this an excellent dataset to explore differences in AA δ13C values among tissues in a wild population. We found the δ13C values of the AANESS glutamic acid, proline, serine, andglycine and the AAESS threonine differed significantly among tissues, indicating possible isotopic discrimination during tissue synthesis. Threonine δ13C values were higher in liver relative to bone collagen and muscle, which may indicate catabolism of threonine for gluconeogenesis, an interpretation further supported by correlations between the δ13C values of threonine and its gluconeogenic products glycine and serine in liver. This intraindividual isotopic variation yielded different ecological interpretations among tissues; for 6/10 of the sea otter individuals analyzed, at least one tissue indicated reliance on a different primary producer source than the other tissues. Our results highlight the importance of gluconeogenesis in a carnivorous marine mammal and indicate that metabolic processes influence AAESS and AANESS δ13C values and multivariate AA δ13C patterns.

Hydroxylysine deficiency, conspicuous skeletal lesions and a strange burial practice in a historical German island population

AbstractEighty‐two complete and partial human skeletons were excavated from a burial site on the island of Borkum in the North Sea. The site covers a period of 400 years from the middle of the 14th century AD until 1720 AD. The position of the skeletons in situ suggests rapid disposal of the bodies. About 75% of the skeletons were male and of a quite robust constitution, and about 60% of all individuals died rather young between 20 and 40 years of age. All male skeletons exhibited particular pathological alterations as a result of the typical daily activities of seafaring men, including severe degenerative joint disease and pronounced osteochondropathies. In juvenile skeletons, these pathologies were accompanied by inflammatory processes without exception. Palaeodietary reconstruction by stable isotope analysis resulted in a protein rich diet, confirming fishing as an important part of the subsistence economy. The juveniles derived substantially more protein from plant food than the adults. Bone collagen amino acid profiles were short of hydroxylysine, suggesting that the conspicuous pathological lesions could also have been caused by a systemic connective tissue disease. Possible etiology of the disease is discussed and related to both the uneven sex ratio of the inhumations and the burial context.

Broad-scale trophic shift in the pelagic North Pacific revealed by an oceanic seabird.

Human-induced ecological change in the open oceans appears to be accelerating. Fisheries, climate change and elevated nutrient inputs are variously blamed, at least in part, for altering oceanic ecosystems. Yet it is challenging to assess the extent of anthropogenic change in the open oceans, where historical records of ecological conditions are sparse, and the geographical scale is immense. We developed millennial-scale amino acid nitrogen isotope records preserved in ancient animal remains to understand changes in food web structure and nutrient regimes in the oceanic realm of the North Pacific Ocean (NPO). Our millennial-scale isotope records of amino acids in bone collagen in a wide-ranging oceanic seabird, the Hawaiian petrel (Pterodroma sandwichensis), showed that trophic level declined over time. The amino acid records do not support a broad-scale increase in nitrogen fixation in the North Pacific subtropical gyre, rejecting an earlier interpretation based on bulk and amino acid specific δ15N chronologies for Hawaiian deep-sea corals and bulk δ15N chronologies for the Hawaiian petrel. Rather, our work suggests that the food web structure in the NPO has shifted at a broad geographical scale, a phenomenon potentially related to industrial fishing.

An overview of methods used for the detection of aquatic resource consumption by humans: Compound-specific delta N-15 analysis of amino acids in archaeological materials

The detection of aquatic resource consumption is important in paleodietary studies as it provides insights into the adaptive strategies of modern humans. Debate as to whether or not it is a recent innovation in hominid evolution also emphasizes its importance to the field. Stable carbon and nitrogen isotope analysis of bulk collagen has been powerful tool for the investigation of ancient human diet, especially for the detection of marine food consumption. However, the nitrogen isotopic composition could significantly vary even within the same animal species depending on environmental conditions, making a dietary reconstruction based on human remains difficult without enough specimens of referential animal prey. Nitrogen isotope analysis of individual amino acids in bone collagen has recently been proposed as a potentially useful technique for the detection of not only marine resources in diets but also freshwater ones. In this paper, we further test its applicability to human populations in a variety of regions and chronological time periods in the world. We found that: (i) irrespective of climatic conditions, nitrogen isotopic compositions of glutamic acid and phenylalanine of terrestrial animals group similarly; (ii) these two amino acids are particularly useful to detect marine or freshwater resource consumption; (iii) a multinomial logistic regression based on some combinations of isotope indicators successfully distinguishes aquatic and terrestrial animals; (iv) detection of aquatic resource consumption by prehistoric humans could be possible based solely on the isotopic compositions of human remains, even in the absence of coeval potential prey. This approach is therefore promising for the paleodietary reconstruction of isolated human remains with few associated faunal remains from archaeological sites.

Refining human palaeodietary reconstruction using amino acid δ15N values of plants, animals and humans

An established method of estimating the trophic level of an organism is through stable isotope analysis of its tissues and those of its diet. This method has been used in archaeology to reconstruct past human diet from the stable nitrogen isotope (δ15N) values of human and herbivore bone collagen. However, this approach, using the 15N-enrichment of human bone collagen δ15N values over associated herbivore bone collagen δ15N values to predict the relative importance of animal protein, relies on the assumptions that: (i) the δ15N values of plants consumed by humans and herbivores are identical, and (ii) the 15N-enrichment between diet and consumer is consistent. Bone collagen amino acid δ15N values have the potential to tackle these uncertainties, as they constrain the factors influencing bone collagen δ15N values. In this study, the δ15N values of glutamic acid and phenylalanine in human and herbivore bone collagen isolates from Neolithic sites in Germany, Greece and Turkey were determined by gas chromatography-combustion-isotope ratio mass spectrometry. The fraction of animal protein in total dietary protein consumed by the humans was estimated by: (i) comparing bulk human and herbivore collagen δ15N values, (ii) comparing bulk human and herbivore collagen and ancient charred cereal grain δ15N values, (iii) comparing human bone collagen δ15NGlutamic acid and δ15NPhenylalanine values, and (iv) comparing δ15NGlutamic acid values of human and herbivore bone collagen and estimated δ15NGlutamic acid values of ancient charred cereal grains. Where determined cereal grain δ15N values are higher than estimated herbivore forage values, estimates of animal protein consumption are significantly lower, emphasising the importance of the plant nitrogen contribution to human bone collagen. This study also highlights the need for further investigation into: (i) the Δ15NConsumer-Diet values of glutamic acid and phenylalanine in terrestrial ecosystems, and (ii) Δ15NGlutamic acid-Phenylalanine values of common plant foods in order to improve the accuracy and more widespread applicability of amino acid-based methods for palaeodietary reconstruction.

Refinement of reconstructed ancient food webs based on the nitrogen isotopic compositions of amino acids from bone collagen: A case study of archaeological herbivores from Tell Ain el-Kerkh, Syria

We determined the stable nitrogen isotopic composition (δ 15 N) of amino acids in bone collagen from samples of three archaeological herbivores (cattle, sheep, and goats), collected from the Tell Ain el-Kerkh Neolithic site in Syria. Bulk collagen δ 15 N values exhibited significant differences between the three species (by up to 3.2‰), and were strongly correlated with those of glycine (R 2 = 0.87), the most abundant amino acid in bone collagen. On the other hand, the δ 15 N values of two other minor amino acids (glutamic acid and phenylalanine) in the different samples were within narrow ranges (0.9‰ and 0.5‰, respectively), and exhibited either weak or no correlation with those of bulk collagen. The trophic position estimated by the δ 15 N values of glutamic acid and phenylalanine (2.0 ± 0.1) is consistent with that of herbivores. These results suggest that the δ 15 N values of bulk bone collagen may vary among herbivores, partly on account of their differing amino acid compositions, whereas the trophic position of different herbivores is faithfully preserved in the δ 15 N values of glutamic acid and phenylalanine.

Evaluation of carnivory in inland Jomon hunter–gatherers based on nitrogen isotopic compositions of individual amino acids in bone collagen

Archaeological studies including stable carbon and nitrogen isotope analyses of bone collagen from human remains have suggested their heavy dependence on terrestrial foods during the Jomon period in the inland central region in Japan. However, it is not easy to quantitatively evaluate the extent of carnivory for archaeological human remains based on the bulk collagen chemistry, because of variable 15N-enrichment factor along the trophic step and background isotopic variations in ecosystems. In order to overcome these problems and more precisely evaluate diets of prehistoric humans who strongly adapted to terrestrial environment, in this study we applied nitrogen isotope analysis of individual amino acids in bone collagen to two inland human populations in the Jomon period. Our results suggest that the two populations were predominantly dependent on the C3-plant-based terrestrial ecosystem and consumed little aquatic resources. Furthermore, their mean trophic positions (2.7 for both cases) are closer to that of the fox (2.8–3.0) rather than those of pure herbivores (2.0–2.2), and show little change over time. These results are the first evidence that inland Jomon populations may have had more carnivorous diets than is traditionally considered.

Variation of bone collagen amino acid δ13c values in archaeological humans and fauna with different dietary regimes: Developing frameworks of dietary discrimination

We present bone collagen amino acid (AA) δ(13)C values for a range of archaeological samples representing four "benchmark" human diet groups (high marine protein consumers, high freshwater protein consumers, terrestrial C(3) consumers, and terrestrial C(4) consumers), a human population with an "unknown" diet, and ruminants. The aim is to establish an interpretive palaeodietary framework for bone collagen AA δ(13)C values, and to assess the extent to which AA δ(13)C values can provide additional dietary information to bulk collagen stable isotope analysis. Results are analyzed to determine the ability of those AAs for which we have a complete set, to discriminate between the diet groups. We show that very strong statistical discrimination is obtained for all interdiet group comparisons. This is often obvious from suitably chosen bivariate plots using δ(113)C values that have been normalized to compensate for interdiet group differences in bulk δ(13)C values. Bi-plots of non-normalized phenylalanine and valine δ(13)C values are useful for distinguishing aquatic diets (marine and freshwater) from terrestrial diets. Our interpretive framework uses multivariate statistics (e.g., discriminant analysis) to optimize the separation of the AA δ(13)C values of the "benchmark"' diet groups, and is capable of accurately assigning external samples to their expected diet groups. With a growing body of AA δ(13)C values, this method is likely to enhance palaeodietary research by allowing the "unknown" diets of populations under investigation to be statistically defined relative to the well-characterized or "known" diets of previously investigated populations.

Dietary reconstruction of prehistoric Japanese populations based on nitrogen isotopic composition of individual amino acids in bone collagen

Dietary reconstruction of prehistoric Japanese populations based on nitrogen isotopic composition of individual amino acids in bone collagen

PP-197 Pyrazinamide-mediated alterations in male rats DNA fragmentation processes, bone collagen amino acid composition, reproductive capability and posterity antenatal and postnatal development

PP-197 Pyrazinamide-mediated alterations in male rats DNA fragmentation processes, bone collagen amino acid composition, reproductive capability and posterity antenatal and postnatal development

Quantitative evaluation of marine protein contribution in ancient diets based on nitrogen isotope ratios of individual amino acids in bone collagen: An investigation at the Kitakogane Jomon site

Nitrogen stable isotopes analysis of individual bone collagen amino acids was applied to archeological samples as a new tool for assessing the composition of ancient human diets and calibrating radiocarbon dates. We used this technique to investigate human and faunal samples from the Kitakogane shell midden in Hokkaido, Japan (5,300-6,000 cal BP). Using compound-specific nitrogen isotope analysis of individual amino acids, we aimed to estimate i) the quantitative contribution of marine and terrestrial protein to the human diet, and ii) the mean trophic level (TL) from which dietary protein was derived from marine ecosystems. Data were interpreted with reference to the amino acid trophic level (TL(AA)) model, which uses empirical amino acid delta(15)N from modern marine fauna to construct mathematical equations that predict the trophic position of organisms. The TL(AA) model produced realistic TL estimates for the Kitakogane marine animals. However, this model was not appropriate for the interpretation of human amino acid delta(15)N, as dietary protein is derived from both marine and terrestrial environments. Hence, we developed a series of relevant equations that considered the consumption of dietary resources from both ecosystems. Using these equations, the mean percentage of marine protein in the Kitakogane human diet was estimated to be 74%. Although this study is one of the first systematic investigations of amino acid delta(15)N in archeological bone collagen, we believe that this technique is extremely useful for TL reconstruction, palaeodietary interpretation, and the correction of marine reservoir effects for radiocarbon dating.

Amino acid δ13C analysis of hair proteins and bone collagen using liquid chromatography/isotope ratio mass spectrometry: paleodietary implications from intra‐individual comparisons

We report a novel method for the chromatographic separation and measurement of stable carbon isotope ratios (delta(13)C) of individual amino acids in hair proteins and bone collagen using the LC-IsoLink system, which interfaces liquid chromatography (LC) with isotope ratio mass spectrometry (IRMS). This paper provides baseline separation of 15 and 13 of the 18 amino acids in bone collagen and hair proteins, respectively. We also describe an approach to analysing small hair samples for compound-specific analysis of segmental hair sections. The LC/IRMS method is applied in a historical context by the delta(13)C analysis of hair proteins and bone collagen recovered from six individuals from Uummannaq in Greenland. The analysis of hair and bone amino acids from the same individual, compared for the first time in this study, is of importance in palaeodietary reconstruction. If hair proteins can be used as a proxy for bone collagen at the amino acid level, this validates compound-specific isotope studies using hair as a model for palaeodietary reconstruction. Our results suggest that a small offset observed in the bulk delta(13)C values of the hair and bone samples may be attributed to two factors: (i) amino acid compositional differences between hair and bone proteins, and (ii) differential turnover rates of the tissues and the amino acid pools contributing to their synthesis. This application proposes that hair may be a useful complementary or alternative source of compound-specific paleodietary information.

Collagen-related abnormalities, reduction in bone quality, and effects of menatetrenone in rats with a congenital ascorbic acid deficiency

In this study, we focused on collagen metabolism as a factor involved in menatetrenone (MK-4)-related improvement in bone quality. Using rats with a congenital ascorbic acid (AA) deficiency, osteogenic disorder Shionogi (ODS) rats, we established a model in which abnormal collagen metabolism reduced bone mechanical properties, and investigated the effects of MK-4. We divided 13-week-old ODS rats into four groups: Pre, AA sufficiency (AA(+)), AA deficiency-control (AA(-)control), and AA deficiency+ MK-4-treated (AA(-)MK-4). MK-4 was given as a dietary supplement (30 mg/kg). At the beginning (pre) and after two, three, and four weeks, seven rats in each group were killed to measure plasma bone metabolism and femoral bone mass data and bone mechanical properties. In the rats killed after four weeks, histomorphometric data of the tibiae, the total amino acid level in bone collagen, and rates of proline and lysine hydroxylation were determined. In the AA(+)group, both the cortical bone mass data and bone mechanical properties were serially increased. However, in the AA(-)control group, the cortical bone mass data were similar for four weeks and the bone mechanical properties decreased after three to four weeks. After four weeks, the total level of amino acids in bone collagen and rates of proline and lysine hydroxylation were significantly lower in the AA(-)control group than in the AA(+)group. MK-4 increased bone mechanical properties after four weeks without influencing cortical bone mass. Simultaneously, it inhibited decreases in the total level of amino acids in collagen (P = 0.017). The rates of proline and lysine hydroxylation were higher in the AA(-)MK-4 group than in the AA(-)control group, but not significantly. These results suggest the level of collagen and abnormalities of hydroxylation are involved in the AA deficiency-related reduction in bone mechanical properties, and that MK-4 improves bone mechanical properties by restoring collagen metabolism.

A novel marine dietary indicator utilising compound-specific bone collagen amino acid δ 13C values of ancient humans

The application of bone collagen stable carbon and nitrogen isotope analysis to human palaeodietary reconstruction in tropical or arid regions is limited by two factors: (i) the overlap in C 4 and high marine protein (HMP) consumer bulk collagen δ 13C values, and (ii) the unpredictability of bulk collagen δ 15N values in regions of extreme aridity (<400 mm of rain per annum). Hence, the identification of HMP consumption among archaeological human populations can be problematic. In an endeavour to identify a more precise marine palaeodietary indicator, a range of collagen samples from archaeological faunal and human bone ( n = 14 and 26, respectively), representing a spectrum of C 3, C 4 and HMP diets, were selected from coastal and near-coastal sites in the Western Cape, South Africa. Samples were subjected to compound-specific stable carbon isotope analysis of their constituent amino acids as trifluoroacetyl-isopropyl (TFA-IP) esters via gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). While human C 4 and HMP consumers were indistinguishable with respect to bulk collagen δ 13C values (−10.9±3.7‰ and −11.7±1.5‰, respectively) they were shown to be readily distinguished based on Δ 13C Glycine-Phenylalanine values (+4.0±1.6‰ and +12.0±1.9‰, respectively). The relationship between HMP consumption and elevated Δ 13C Glycine-Phenylalanine values was verified by: (i) the similarly elevated values exhibited by marine species when compared to terrestrial faunal species (+12.5±0.9‰ and +3.2±4.2‰, respectively), and (ii) the strong correlation observed between human Δ 13C Glycine-Phenylalanine and bulk collagen δ 15N values ( R 2 = 0.83 , p < 0.001 ; n = 26 ), the latter being a well-documented marine dietary indicator. It was concluded that Δ 13C Glycine-Phenylalanine values offer considerable potential as indicators of HMP consumption and a valuable substitute for bone collagen δ 15N values in arid regions where bulk δ 15N values are unpredictable.

The isotopic composition of carbon and nitrogen in individual amino acids isolated from modern and fossil proteins

Pigs were reared in laboratory pens on controlled diets that consisted of either 100% C 3 plants or 100% C 4 plants. Carbon and nitrogen isotopic compositions of the diets, and the resulting pig products, purified collagen and muscle tissue, were measured to determine isotopic fractionation during growth and metabolism. Total collagen from pigs grown on C 3 diets was enriched in 13C by 3·2‰ and in 15N by 2·2‰, whereas that from pigs reared on C 4 diets was enriched in 13C by 1·4‰ and in 15N by 2·3‰. In addition, fractionation between pigs and their diets was determined at the molecular level on individual amino acids separated by ion exchange chromatography. The carbon isotopic compositions of separated amino acids from the C 3 and C 4 diets were transferred to amino acids in bone collagen. For nitrogen, the isotopic compositions of all nonessential amino acids were enriched in 15N relative to those amino acids in the diet. Threonine, an essential amino acid, behaved oppositely, in that its isotope ratio (δ 15N) was depleted by an average of 6‰ from the δ 15N of the whole collagen. Similar isotopic patterns were analysed in collagenous amino acids extracted from field specimens that included both herbivores and carnivores; marine animals and terrestrial animals; and C 3 and C 4 feeders. Amino acids from two fossil bones, a bison (4500 years old) and a whale (70,000 years old), recorded the same isotopic signals as modern collagen. The ubiquity of these isotopic patterns at the molecular level suggests that distinct biochemical mechanisms control the metabolism of amino acids in animals rather than random synthesis.

Role of Copper in Collagen Cross-linking and Its Influence on Selected Mechanical Properties of Chick Bone and Tendon

Day-old White Leghorn cockerels were fed skim-milk-based diets containing 0, 2, 4, 8, or 16 ppm added copper as CuSO4 for 21 days. Lysyl oxidase was extracted from bone and tendon. The recovery of activity from the extracts was linearly correlated with dietary copper (r > 0.90). Tests of the mechanical properties of tendon and bone indicated that tendon viscoelasticity (as measured by stress-relaxation) may not be significantly influenced by copper, whereas the ultimate torsional strength of bone is markedly decreased when the dietary copper level is below 1 ppm. Furthermore the torsional fracture characteristics of bone from copper-deficient birds demonstrated a lack of plastic deformation prior to failure that was normally seen in bone from control birds. The change in the mechanical properties of bone from copper-deficient birds appeared to be related to a decrease in the amounts of dihydroxylysinonorleucine and other lysine-derived cross-linking amino acids in bone collagen. The data indicate that the requirement for optimal growth of chicks fed skim-milk-based diets is 6–8 ppm copper, however, the requirement for normal cross-link formation in bone is less than 2 ppm copper.copper cross-linking amino acids biomechanics bone collagen tendon collagen

Vinca Skeletons Studied in Situ at the Gomolava Site, Yugoslavia

Previous articleNext article No AccessResearch ConclusionsVinca Skeletons Studied in Situ at the Gomolava Site, YugoslaviaSrboljub ZivanovicSrboljub Zivanovic Search for more articles by this author PDFPDF PLUS Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinkedInRedditEmail SectionsMoreDetailsFiguresReferencesCited by Current Anthropology Volume 18, Number 3Sep., 1977 Sponsored by the Wenner-Gren Foundation for Anthropological Research Article DOIhttps://doi.org/10.1086/201938 Views: 3Total views on this site Citations: 2Citations are reported from Crossref Copyright 1977 The Wenner-Gren Foundation for Anthropological ResearchPDF download Crossref reports the following articles citing this article:Đurica Grga, Ilija Mikić, Bogdan Lisul, Tamara Zlopaša, Bojan Dželetović Original article. Dental and Jaws Status in Pre-historic Human Population of the Gomolava Site, Stomatoloski glasnik Srbije 64, no.11 (Mar 2017): 24–31.https://doi.org/10.1515/sdj-2017-0003Noah V. Honch, James S.O. McCullagh, Robert E.M. Hedges Variation of bone collagen amino acid δ 13 c values in archaeological humans and fauna with different dietary regimes: Developing frameworks of dietary discrimination, American Journal of Physical Anthropology 148, no.44 (May 2012): 495–511.https://doi.org/10.1002/ajpa.22065