Changes in the intramolecular stable carbon isotope ratios with age of the European cave bear (Ursus spelaeus)

Changes with age in the diet and metabolism of the extinct European cave bear (Ursus spelaeus) from the Divje Babe archaeological site in northwestern Slovenia were investigated by comparing the stable carbon isotope ratio of whole bone collagen with that of the peptide-bonded carboxyl carbons in collagen. These carboxyl carbons were selectively released by decarboxylation of the collagen hydrolysate with ninhydrin (2,2-dihydroxy-1,3-indanedione). The stable carbon isotope ratio increased with age for both the peptide-bonded and other carbons. Although only one-third of the carbon atoms in collagen are peptide-bonded, they account for most of the observed change with age of the stable carbon isotope ratio of the whole collagen. Stable nitrogen isotope ratios decreased with age and were negatively correlated with changes in the stable carbon isotope ratios. These observations support previous investigations on the unique metabolism of hibernating bears.     

Stable isotope evidence for 1500 years of human diet at the city of York, UK

We present here the results of a large-scale diachronic palaeodietary (carbon and nitrogen isotopic measurements of bone collagen) study of humans and animals from a single site, the city of York (U.K.), dating from the Roman period to the early 19th century. The human sample comprises 313 burials from the cemeteries of Trentholme Drive and Blossom Street (Roman), Belle Vue House (Anglo-Saxon), Fishergate (High and Later Medieval), and All Saints, Pavement (Later and Post-Medieval). In addition, 145 samples of mammal, fish and bird bone from the sites of Tanner Row and Fishergate were analyzed. The isotope data suggest dietary variation between all archaeological periods, although the most significant change was the introduction of significant quantities of marine foods in the Medieval periods. These are first evident in the diet of a small group of individuals from the High Medieval cemetery at Fishergate, although they were consumed almost universally in the following periods. The human isotope values are also remarkable due to unusually elevated delta(15)N ratios that are not sufficiently explained by the comparably small enrichment in (13)C that accompanies them. We discuss the possible reasons behind this and the archaeological significance of the data set.     

Technical note: interpreting stable carbon isotopes in human tooth enamel: an examination of tissue spacings from South Africa

Stable isotope analysis of skeletal tissues is widely used in archeology and paleoanthropology to reconstruct diet. In material that is poorly preserved or very old, the tissue of choice is frequently tooth enamel, since this is less susceptible to diagenesis. The relationships between carbon isotope ratios in tooth enamel (δ(13) C(enamel) ), bone collagen (δ(13) C(collagen) ), and bone apatite (δ(13) C(bone apatite) ) are, however, not well understood. To elucidate these, we have measured all three indicators in archeological humans from the western and southern Cape coastal regions of South Africa. The correlation between δ(13) C(enamel) and δ(13) C(collagen) is good (R(2) = 0.71 if two outliers are excluded, n = 79). The correlation between δ(13) C(enamel) and δ(13) C(bone apatite) is weaker (R(2) = 0.37, n = 33) possibly due to bone diagenesis. No systematic offset between δ(13) C(bone apatite) and δ(13) C(enamel) was observed in this sample of archeological humans. Intertooth comparisons of δ(13) C(enamel) in three individuals showed little variation, despite the different ages of crown formation. Carbon isotope ratios in both enamel and bone collagen are good proxies for δ(13) C(diet) .     

Degradation of biomolecules in bones: effects of the biological forensics as an example of the stability of isotope ratios in collagen

Modern bone samples were experimentally degraded by incubation into water at increased temperature and examined in terms of their collagen content, the stable C and N isotopic ratios, and the molar C/N ratio. The same analyses were carried out with archaeological human bone of varying age (300 up to 8000 years). The experimentally degraded samples exhibited changes of the collagen's integrity, which influence the stable isotope ratios. In the case of the archaeological material, a correlation between stable delta13C- and delta15N-values and collagen content could be demonstrated. The molar C:N ratio was no suitable criterion for the assessment of the state of preservation of extractable collagen.     

Bone protects proteins over thousands of years: extraction, analysis, and interpretation of extracellular matrix proteins in archeological skeletal remains

In a good state of preservation, bone conserves the entire protein pattern of extracellular bone matrix proteins over thousands of years. The quality of the profiles of matrix proteins isolated from ancient bones (ranging from the pre-Pottery Neolithic Phase to Early Modern Times from different archaeological sites in different geographical areas), separated by electrophoresis, is as good as those from recent bones. Molecules arising from collagenous proteins (e.g., collagen type I), from the noncollagenous group (e.g., osteonectin), and from the immune system (e.g., immunoglobulin G) were identified in Western blots by specific antibodies. A comparative study of the immunoglobulin G content of the bones of five prehistoric children showed the lowest immunoglobulin G content in a child who suffered from chronic scurvy. Ancient bone proteins were also separated by two-dimensional electrophoresis. This technique makes fractionation of the complex protein mixtures of extracellular bone matrix more reproducible. Bone retains a chemical memory of earlier metabolic stimuli in its configuration of collagenous and noncollagenous proteins. In combination with the results of the microscopic examination of ancient bone, it should be possible to obtain more reliable information on the history and the evolution of diseases, based on analysis of intact proteins.     

Eating between the Lines: Mississippian Migration and Stable Carbon Isotope Variation in Fort Ancient Populations

ABSTRACT   Appreciating variation along the edges of traditional archaeological Culture Historical boundaries requires close consideration of social contexts associated with culture contact. We focus on dietary variation as a function of these concerns through a case study of Fort Ancient populations who, on average, consumed lower quantities of maize than their Mississippian neighbors as determined by stable carbon isotope ratios of bone collagen. However, this dichotomy is not as rigid as initially thought, with some Fort Ancient burials producing stable carbon isotope ratios similar to Mississippian cases. Detailed investigation of internal variation of carbon isotopes for human burials at the SunWatch site provides evidence that contact included small-scale Mississippian migrations to Fort Ancient sites. The main conclusion is that variation in diet and archaeological context can be a useful approach for examining prehistoric migration.     

Victims and survivors: Stable isotopes used to identify migrants from the Great Irish Famine to 19th century London

Historical evidence documents mass migration from Ireland to London during the period of the Great Irish Famine of 1845–52. The rural Irish were reliant on a restricted diet based on potatoes but maize, a C₄ plant, was imported from the United States of America in 1846–47 to mitigate against Famine. In London, Irish migrants joined a population with a more varied diet. To investigate and characterize their diet, carbon and nitrogen isotope ratios were obtained from bone collagen of 119 and hair keratin of six individuals from Lukin Street cemetery, Tower Hamlets (1843–54), and bone collagen of 20 individuals from the cemetery at Kilkenny Union Workhouse in Ireland (1847–51). A comparison of the results with other contemporaneous English populations suggests that Londoners may have elevated δ¹⁵N compared with their contemporaries in other cities. In comparison, the Irish group have lower δ¹⁵N. Hair analysis combined with bone collagen allows the reconstruction of perimortem dietary changes. Three children aged 5–15 years from Kilkenny have bone collagen δ¹³C values that indicate consumption of maize (C₄). As maize was only imported into Ireland in quantity from late 1846 and 1847, these results demonstrate relatively rapid bone collagen turnover in children and highlight the importance of age‐related bone turnover rates, and the impact the age of the individual can have on studies of short‐term dietary change or recent migration. Stable light isotope data in this study are consistent with the epigraphic and documentary evidence for the presence of migrants within the London cemetery. Am J Phys Anthropol, 2013. © 2012 Wiley Periodicals, Inc.     

Density fractionation of forest soils: methodological questions and interpretation of incubation results and turnover time in an ecosystem context

Soil organic matter (SOM) is often separated by physical means to simplify a complex matrix into discrete fractions. A frequent approach to isolating two or more fractions is based on differing particle densities and uses a high density liquid such as sodium polytungstate (SPT). Soil density fractions are often interpreted as organic matter pools with different carbon (C) turnover times, ranging from years to decades or centuries, and with different functional roles for C and nutrient dynamics. In this paper, we discuss the development and mechanistic basis of common density-based methods for dividing soil into distinct organic matter fractions. Further, we directly address the potential effects of dispersing soil in a high density salt solution on the recovered fractions and implications for data interpretation. Soil collected from forested sites at H. J. Andrews Experimental Forest, Oregon and Bousson Experimental Forest, Pennsylvania was separated into light and heavy fractions by floatation in a 1.6 g cm⁻³ solution of SPT. Mass balance calculations revealed that between 17% and 26% of the original bulk soil C and N content was mobilized and subsequently discarded during density fractionation for both soils. In some cases, the light isotope was preferentially mobilized during density fractionation. During a year-long incubation, mathematically recombined density fractions respired ∼40% less than the bulk soil at both sites and light fraction (LF) did not always decompose more than the heavy fraction (HF). Residual amounts of tungsten (W) present even in well-rinsed fractions were enough to reduce microbial respiration by 27% compared to the control in a 90-day incubation of O_a material. However, residual W was nearly eliminated by repeated leaching over the year-long incubation, and is not likely the primary cause of the difference in respiration between summed fractions and bulk soil. Light fraction at Bousson, a deciduous site developed on Alfisols, had a radiocarbon-based mean residence time (MRT) of 2.7 or 89 years, depending on the interpretation of the radiocarbon model, while HF was 317 years. In contrast, both density fractions from H. J. Andrews, a coniferous site developed on andic soils, had approximately the same MRT (117 years and 93 years for LF and HF). At H. J. Andrews the organic matter lost during density separation had a short MRT (19 years) and can account for the difference in respired CO₂ between the summed fractions and the bulk soil. Recognition and consideration of the effects of the density separation procedure on the recovered fractions will help prevent misinterpretation and deepen our understanding of the specific role of the recovered organic matter fractions in the ecological context of the soil studied.     

Collagen and non-collagenous proteins in different mineralization stages of human femur

Mineralized tissues exhibit varying degrees of mineralization in different areas within the same bone. Using the technique of density gradient fractionation, bone powder from the diaphysis of human femur has been separated in different fractions corresponding to the degree of mineralization. Isolated bone fractions were analysed for their content in collagen and non-collagenous proteins. The results showed marked differences between compact and spongy bones, this latter containing higher proportions of little mineralized bone particles than the former (p less than 0.01). The ethylenediaminetetra-acetic acid (EDTA) extractability and the bone matrix size decreased relative to the decrease in specific gravity of bone particles. Among the matrix components of different fractions, sialoprotein consistently increased with the increase in specific gravity while proteoglycan decreased in reverse manner to the increase in collagen. However, in the most mineralized fraction (specific gravity: 2.33 g/cm3), the proteoglycan amount increased while collagen decreased. In conclusion, this study of bone maturation in human femur confirms the suitability of the technique of density gradient fractionation in the studies of bone matrix-mineral interactions. Apart from the fraction with the highest specific gravity, the analytical results obtained in fractions are similar to those observed in age-related bone changes, suggesting that the increase in mineralization degree of bone particles may be related to their age.     

The Ice Man's diet as reflected by the stable nitrogen and carbon isotopic composition of his hair.

Establishing the diets of ancient human populations is an integral component of most archaeological studies. Stable isotope analysis of well-preserved bone collagen is the most direct approach for a general assessment of paleodiet. However, this method has been limited by the scarcity of well-preserved skeletal materials for this type of destructive analysis. Hair is preserved in many burials, but is often overlooked as an alternative material for isotopic analysis. Here we report that the stable carbon and nitrogen isotope values for the hair of the 5200 year-old Ice Man indicates a primarily vegetarian diet, in agreement with his dental wear pattern. Whereas previous investigations have focused on bone collagen, the stable isotope composition of hair may prove to be a more reliable proxy for paleodiet reconstruction, particularly when skeletal remains are not well preserved and additional archaeological artifacts are unavailable.     

Palaeodiet reconstruction in a woman with probable celiac disease: A stable isotope analysis of bone remains from the archaeological site of Cosa (Italy)

Stable isotope analysis in the reconstruction of human palaeodiets can yield clues to early human subsistence strategies, origins and history of farming and pastoralist societies, and intra‐ and intergroup social differentiation. In the last 10 years, the method has been extended to the pathological investigation. Stable isotope analysis to better understand a diet‐related disease: celiac disease in ancient human bones was carried out. To do this, we analyzed the nitrogen and carbon isotopic composition of human (n = 37) and faunal (n = 8) bone remains from the archaeological site of Cosa at Ansedonia, on the Tyrrhenian coast near Orbetello (Tuscany), including the skeletal remains of a young woman (late 1st century–early 2nd century Common Era [CE]) with morphological and genetic features suggestive of celiac disease. We compared the young woman's isotopic data with those of other individuals recovered at the same site but from two later time periods (6th century CE; 11–12th century CE) and with literature data from other Italian archaeological sites dating to the same period. Her collagen δ¹³C and δ¹⁵N values differed from those of the samples at the same site, and from most but not all of the contemporary sites. Although the woman's diet appears distinct, chronic malnutrition resulting from severe malabsorption of essential nutrients due to celiac disease may have affected the isotopic composition of her bone collagen. Am J Phys Anthropol 154:349–356, 2014. © 2014 Wiley Periodicals, Inc.     

Modern macaque dietary heterogeneity assessed using stable isotope analysis of hair and bone

Dietary variability might have been a major factor in the dispersal and subsequent persistence of the genus Macaca in both tropical and temperate areas. Macaques are found from northern Africa to Japan, yet there have been few systematic attempts to compare diets between different modern populations. Here we have taken a direct approach and sampled museum-curated tissues (hair and bone) of Macaca mulatta (rhesus macaques) for carbon and nitrogen stable isotope dietary analyses. Samples from India, Vietnam, and Burma (Myanmar) were taken, representing both tropical and temperate populations. The δ¹³C values obtained from hair show that the temperate macaques, particularly those from Uttar Pradesh, have a δ¹³C signature that indicates at least some use of C₄ resources, while the tropical individuals have a C₃-based diet. However, δ¹³C values from bone bioapatite indicate a C₃-based diet for all specimens and they do not show the C₄ usage seen in the hair of some animals, possibly because bone represents a much longer turnover period than that of hair. The results of δ¹⁵N analyses grouped animals by geographic region of origin, which may be related to local soil nitrogen values. The greatest variation in δ¹⁵N values was seen in the specimens from Burma, which may be partly due to seasonality, as specimens were collected at different times of year. We also investigated the relationship between the hair, bone collagen, and bone bioapatite δ¹³C results, and found that they are highly correlated, and that one tissue can be used to extrapolate results for another. However, our results also suggest that hair may pick up discrete feeding traces (such as seasonal usage), which are lost when only bone collagen and bioapatite are examined. This has important implications for dietary reconstructions of archaeological and paleontological populations.     

猪骨胶原蛋白肽缓解高脂饮食诱导小鼠肝脏氧化应激的基因芯片分析

本研究探讨猪骨胶原蛋白肽降血脂作用和抑制高脂饮食诱导肝脏抗氧化应激的作用机理。40只小鼠分为5组,分别饲喂6 w正常日粮、高脂日粮、添加1.6%钙的高脂日粮、添加1%胶原蛋白肽的高脂日粮和添加1.6%钙+1%胶原蛋白肽的高脂日粮。研究发现:与正常组相比,高脂日粮小鼠血脂和体重明显升高。1.6%钙处理可抑制体重过度升高,1%胶原蛋白肽和高钙可协同抑制调节血脂,防止体重升高。高脂主要通过影响肝脏生理节律和过氧化物酶体增生物激活受体信号通路引起脂代谢异常,高钙和骨胶原蛋白肽可显著抑制肝脏氧化应激,抑制生理节律紊乱。     

Collagen turnover in the adult femoral mid-shaft: modeled from anthropogenic radiocarbon tracer measurements

We have measured the (14)C content of human femoral mid-shaft collagen to determine the dynamics of adult collagen turnover, using the sudden doubling and subsequent slow relaxation of global atmospheric (14)C content due to nuclear bomb testing in the 1960s and 1970s as a tracer. (14)C measurements were made on bone collagen from 67 individuals of both sexes who died in Australia in 1990-1993, spanning a range of ages at death from 40 to 97, and these measurements were compared with values predicted by an age-dependent turnover model. We found that the dataset could constrain models of collagen turnover, with the following outcomes: 1) Collagen turnover rate of females decreases, on average, from 4%/yr to 3%/yr from 20 to 80 years. Male collagen turnover rates average 1.5-3%/yr over the same period. 2) For both sexes the collagen turnover rate during adolescent growth is much higher (5-15%/yr at age 10-15 years), with males having a significantly higher turnover rate than have females, by up to a factor of 2. 3) Much of the variation in residual bomb (14)C in a person's bone can be attributed to individual variation in turnover rate, but of no more than about 30% of the average values for adults. 4) Human femoral bone collagen isotopically reflects an individual's diet over a much longer period of time than 10 years, including a substantial portion of collagen synthesised during adolescence.     

Quantitative Reconstruction of Weaning Ages in Archaeological Human Populations Using Bone Collagen Nitrogen Isotope Ratios and Approximate Bayesian Computation

BackgroundNitrogen isotope analysis of bone collagen has been used to reconstruct the breastfeeding practices of archaeological human populations. However, weaning ages have been estimated subjectively because of a lack of both information on subadult bone collagen turnover rates and appropriate analytical models.MethodologyTemporal changes in human subadult bone collagen turnover rates were estimated from data on tissue-level bone metabolism reported in previous studies. A model for reconstructing precise weaning ages was then developed using a framework of approximate Bayesian computation and incorporating the estimated turnover rates. The model is presented as a new open source R package, WARN (Weaning Age Reconstruction with Nitrogen isotope analysis), which computes the age at the start and end of weaning, ¹⁵N-enrichment through maternal to infant tissue, and value of collagen synthesized entirely from weaning foods with their posterior probabilities. The model was applied to 39 previously reported Holocene skeletal populations from around the world, and the results were compared with weaning ages observed in ethnographic studies.ConclusionsThere were no significant differences in the age at the end of weaning between the archaeological (2.80±1.32 years) and ethnographic populations. By comparing archaeological populations, it appears that weaning ages did not differ with the type of subsistence practiced (i.e., hunting–gathering or not). Most of -enrichment (2.44±0.90‰) was consistent with biologically valid values. The nitrogen isotope ratios of subadults after the weaning process were lower than those of adults in most of the archaeological populations (−0.48±0.61‰), and this depletion was greater in non-hunter–gatherer populations. Our results suggest that the breastfeeding period in humans had already been shortened by the early Holocene compared with those in extant great apes.     

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.     

Processing of prehistoric bones for isotopic analysis and the meaning of collagen C/N ratios in the assessment of diagenetic effects

Diagenetic shifts in the isotopic composition of collagen in prehistoric bones still remain a big problem in the reconstruction of ancient diets by stable carbon and nitrogen isotope analysis. Recently,DeNiro (1985) suggested the measurement of bone collagen C/N ratios as a means of estimating substantial alterations of stable isotope ratios. Bones with collagen C/N ratios lying within a range between 2.9 and 3.6 should have isotopic properties quite close to thein vivo conditions. It can be demonstrated that the C/N ratios are varying considerably with the duration of acid hydrolyzation of the bone samples. Even small changes of the hydrolyzation time cause shifts in the C/N ratios large enought to produce values far outside the range worked out byDeNiro. Besides, our experiments led us to recommend a hydrolyzation at reducing conditions.     

百岁以上老人跟骨骨密度和骨强度的相关性分析

目的研究广西巴马百岁以上女性老人的年龄、身高、体重、BMI与跟骨骨密度、骨强度之间的关系。方法采用定量超声骨密度测定仪,对广西巴马16名百岁以上女性老人的跟骨骨密度、骨强度进行测定,并记录相应年龄,同时行人工测量身高、体重、BMI指数,应用SPSS16.0软件分别计算年龄、身高、体重、BMI与跟骨骨密度、骨强度的pearson的相关系数。结果年龄、身高、体重、BMI均与百岁以上女性老人的跟骨骨密度、骨强度有相关关系,但相关性均无显著性(P0.05)。其中年龄、身高、体重与跟骨骨密度、骨强度呈负相关关系;BMI与跟骨骨密度、骨强度呈正相关关系,但相关性均无显著性(P0.05)。结论年龄、身高、体重、BMI均是广西巴马百岁以上女性老人骨密度、骨强度的影响因素,但相关性均无显著性,或需探讨新的评价指标衡量百岁以上女性老人的骨密度及骨强度,为百岁以上女性老人早期预防和诊断骨质疏松提供科学依据。     

Social structures and social relations--an archaeological and anthropological examination of three early Medieval separate burial sites in Bavaria

As part of an interdisciplinary cooperation, three early medieval separated burial sites from central Bavaria were subjected to archaeological and anthropological examinations. Separated burial sites are probably an expression of social stratification. This paper focuses on two characteristic aspects of these separated burial sites: (1) structure of the site, and (2) biological manifestation of a social upper class. The separated burial grounds Etting/Sandfeld (31 individuals), Grossmehring (44 individuals) and Kelheim (43 individuals), all located in southern Bavaria, were analyzed. Sex and age at death of all individuals were determined, and dietary behaviour was reconstructed by means of carbon and nitrogen isotope ratios in bone collagen. Local conditions such as climate, soil conditions or the intensity of agriculture can provoke significant variations in plant delta15N-values, which could lead to a shift of the baseline values of the corresponding trophic web. To facilitate the comparison of isotopic data from different sites, delta15N values of cattle bones were taken as a reference for the human data (presuming the diet to consumer chain). The results of dietary reconstruction indicate that the populations enjoyed very good living conditions with a primarily animal protein-based diet. Furthermore, the isotope analysis revealed more detailed indications for certain individuals regarding their social status. Some individuals can even be appointed as possible chiefs of the population, since anthropological and archaeological interpretations were in total agreement: It was possible to identify persons of a higher social status based on the structure of the burial site, grave goods and the isotopic analysis.     

Diet and dental caries among later stone age inhabitants of the Cape Province, South Africa.

Stable carbon isotope measurements and incidences of dental caries are presented for three groups of prehistoric human skeletons from different regions of the Cape Province, South Africa. The isotopic analyses of bone collagen demonstrate the importance of marine foods in the diet and vary through time, across space, and according to sex. The incidence of dental caries ranges from 0% among heavily marine-dependent individuals from the south-western Cape coast, to 17.7% among skeletons from an archaeological site on the south coast. The extremely high incidence of caries in a hunter-gatherer population may be related to lack of fluoride in the water.