Combining elemental and immunochemical analyses to characterize diagenetic alteration patterns in ancient skeletal remains.

Bones and teeth are biological archives, but their structure and composition are subjected to alteration overtime due to biological and chemical degradation postmortem, influenced by burial environment and conditions. Nevertheless, organic fraction preservation is mandatory for several archeometric analyses and applications. The mutual protection between biomineral and organic fractions in bones and teeth may lead to a limited diagenetic alteration, promoting a better conservation of the organic fraction. However, the correlation between elemental variations and the presence of organic materials (e.g., collagen) in the same specimen is still unclear. To fill this gap, chemiluminescent (CL) immunochemical imaging analysis has been applied for the first time for collagen localization. Then, Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS) and CL imaging were combined to investigate the correlation between elemental (i.e., REE, U, Sr, Ba) and collagen distribution. Teeth and bones from various archeological contexts, chronological periods, and characterized by different collagen content were analyzed. Immunochemical analysis revealed a heterogeneous distribution of collagen, especially in highly degraded samples. Subsequently, LA-ICP-MS showed a correlation between the presence of uranium and rare earth elements and areas with low amount of collagen. The innovative integration between the two methods permitted to clarify the mutual relation between elemental variation and collagen preservation overtime, thus contributing to unravel the effects of diagenetic alteration in bones and teeth.

Near-infrared hyperspectral imaging (NIR-HSI) and normalized difference image (NDI) data processing: An advanced method to map collagen in archaeological bones.

In the present study, an innovative and highly efficient near-infrared hyperspectral imaging (NIR-HSI) method is proposed to provide spectral maps able to reveal collagen distribution in large-size bones, also offering semi-quantitative estimations. A recently introduced method for the construction of chemical maps, based on Normalized Difference Images (NDI), is declined in an innovative approach, through the exploitation of the NDI values computed for each pixel of the hyperspectral image to localize collagen and to extract information on its content by a direct comparison with known reference samples. The developed approach addresses an urgent issue of the analytical chemistry applied to bioarcheology researches, which rely on well-preserved collagen in bones to obtain key information on chronology, paleoecology and taxonomy. Indeed, the high demand for large-sample datasets and the consequent application of a wide variety of destructive analytical methods led to the considerable destruction of precious bone samples. NIR-HSI pre-screening allows researchers to properly select the sampling points for subsequent specific analyses, to minimize costs and time and to preserve integrity of archaeological bones (which are available in a very limited amount), providing further opportunities to understand our past.

[RECONSTRUCTING THE LIFE OF AN UNKNOWN MAN--INTERDISCIPLINARY APPROACH].

The remains of a tomb were discovered below the foundations of the Roman church of "Majka Bozja Gorska" in Lobor in 2002. It was a vaulted tomb carved from a single stone. The narthex of ancient Christian and pre-roman churches often served as the resting place of religious, secular and noble dignitaries, who were credited for founding the church at the time. By anthropological methods, the age, gender, height, pathological changes, tendon and ligament hinges and degree of development were able to be analysed. The stable isotopes of Carbon-13, Nitrogen-15 and Oxygen-18 were analysed and expressed as ��-values according to the PDB standard. Dental analysis showed a build-up of tartar on the buccal side, with 5 teeth missing from the dentition post-mortem and a partial lowering and accretion of the alveolar walls of the 2nd molar space secondary to tooth loss. The remains of the person from grave 107 show that the person was approximately 30-35 years of age, confirmed by microscopical analysis of the bone samples. Almost all of the cranial sutures were exposed with the exception of the saggital suture which was closed at the S3 position, while the transverse palatal suture was in the initial stages of closure on the lateral sides. Analysis of the iliac bone showed visible changes on the bone. The surface had retained its youthful appearance however with a microporous and pronounced granular structure and only a slight apical and marginal activity. The transverse striations are still apparent, but no longer along the entirety of the bone surface. Anthropometric analysis according to cranial cavity indexes, showed a cranium or relatively large volume (aristencephalic in type), long, narrow and medium in height. The index values were used in facial reconstruction. Analysis of the development of tendon and ligament hinges of the preserved anatomical elements, gave further information about the morphology and physical activity of the person. The hinge sites of the brachial and antebrachial muscles showed a significant development and these enthesopathies would be consistent with a strenuous physical activity, especially at the biceps tendon and shoulder girdle, portraying an anterior elevation, cubital flexion, pronation, circumduction and fixation. It can be said that the person predominantly used the left arm. The developed muscle hinges, show common pronatory and supinatory movements and presume a routine swinging motion of an object above the head or in front of the person. The left tibia was suggestive of the height and male gender of the person. According to Pearson's regression formula, for the length of the brachium, radius and tibia, it can be reasoned that the person was approximately 169 cm tall. The upper third of the L3 vertebra shows a Schmorl's hernia which can be secondary to strong anteroposterior and lateral flexion of the spine. On the supero-lateral surface of the left patella, there is a Messeri sign, suggestive of a long and repeated flexion of the knee (kneeling). The molars of the lower and especially upper jaw show signs of paradontosis and a slight abrasion of the first degree according to Gustafson. The secondary dentin can be seen on the incisors, canines and molars. The bone impression of the left temporal bone, was oval in shape and measured 12x9 mm. It was probably and old, healed wound, possibly consequential to a fall or strike from a sharp object. The left tibia, showed a treated but still active periostitis. Analysis of the stable isotopes (C, N, O) shows an increased consumption of vegetables, common to a moderately warm and dry climate (millet). The person had access to water of a good quality, possibly from a well and did not eat fish or drink wine. We do not know whether the person from grave 107 was a member of the clergy or nobility, but was most likely a member of the social elite.

C4 -consumers in southern Europe: the case of Friuli V.G. (NE-Italy) during early and central Middle Ages.

Isotope variations were studied in necropolises of the early (6th to 7th century CE) and central (10th to 11th century CE) medieval period located in Fruili-Venezia Giulia (Northeastern Italy). The two periods each shortly followed two great barbarian invasions that changed the politics and economy of Italy: the arrivals of Langobards in 578 CE and the Hungarian incursions from the end of the 9th to the first half of the 10th century. These events had a tragic effect on the economy of Friuli-Venezia Giulia: severe depopulation and the partial abandonment of the countryside with fall of agricultural production.

Last Glacial mammals in South America: a new scenario from the Tarija Basin (Bolivia).

The chronology, sedimentary history, and paleoecology of the Tarija Basin (Bolivia), one of the richest Pleistocene mammalian sites in South America, are revised here based on a multidisciplinary study, including stratigraphy, sedimentology, geomorphology, paleontology, isotope geochemistry, and (14)C geochronology. Previous studies have indicated a Middle Pleistocene age for this classic locality. We have been able to obtain a series of (14)C dates encompassing all the fossil-bearing sequences previously studied in the Tarija Basin. The dated layers range in age from about 44,000 to 21,000 radiocarbon years before present (BP), indicating that the Tarija fauna is much younger than previously thought. Glacial advances correlated to marine isotopic stages (MIS) 4 and 2 (ca. 62 and 20 ka BP, respectively) are also documented at the base and at the very top of the Tarija-Padcaya succession, respectively, indicating that the Bolivian Altiplano was not dry but sustained an ice cap during the Last Glacial Maximum. The results of this multidisciplinary study enable us to redefine the chronological limits of the Tarija sequence and of its faunal assemblage and to shift this paleontological, paleoclimatological, and paleoecological framework to the time interval from MIS 4 to MIS 2.

Daily energy expenditure in free-living conditions in obese and non-obese children: comparison of doubly labelled water (2H2(18)O) method and heart-rate monitoring.

OBJECTIVE: To compare the heart-rate monitoring with the doubly labelled water (2H2(18)O) method to estimate total daily energy expenditure in obese and non-obese children. DESIGN: Cross sectional study of obese and normal weight children. SUBJECTS: 13 prepubertal children: six obese (4M, 2F, 9.1 +/- 1.5 years, 47.3 +/- 9.7 kg) and seven non-obese (3M, 4F, 9.3 +/- 0.6 years, 31.8 +/- 3.2 kg). MEASUREMENTS: Total daily energy expenditure was assessed by means of the doubly labelled water method (TEEDLW) and of heart-rate monitoring (TEEHR). RESULTS: TEEHR was significantly (P < 0.05) higher than TEEDLW in obese children (9.47 +/- 0.84 MJ/d vs 8.99 +/- 0.63 MJ/d) whereas it was not different in non-obese children (8.43 +/- 2.02 MJ/d vs 8.42 +/- 2.30 MJ/d, P = NS). The difference of TEE assessed by HR monitoring in the obese group averaged 6.2 +/- 4.7%. At the individual level, the degree of agreement (difference between TEEHR and TEEDLW +/- 2s.d.) was low both in obese (-0.36, 1.32 MJ/d) and in non-obese children (-1.30, 1.34 MJ/d). At the group level, the agreement between the two methods was good in nonobese children (95% c.i. for the bias:-0.59, 0.63 MJ/d) but not in obese children (0.04, 0.92 MJ/d). Duration of sleep and energy expenditure during resting and physical activity were not significantly different in the two groups. Patterns of heart-rate (or derived energy expenditure) during the day-time were similar in obese and non-obese children. CONCLUSION: The HR monitoring technique provides an estimation of TEE close to that assessed by the DLW method in non-obese prepubertal children. In comparison with DLW, the HR monitoring method yields a greater TEE value in obese children.