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The analysis of traces of injuries can be difficult in cases of charred human remains since the alteration and fragmentation are high. The aim of this study is to explore the use of X-Ray Fluorescence (XRF) technique as a screening tool for detecting and analyzing gunshot residues (GSR) on cremated and highly fragmented materials, as it is a technique that allows for fast qualitative investigations without altering the sample or requiring sample preparation. The study was carried out on two steps: firstly, on completed skeletonized bones to verify if GSR survive to burning; secondly, we considered a more realistic situation, in which soft tissues were present before the shooting. To this aim, nine adult bovine ribs, four retaining soft tissue, five completely skeletonized, were subjected to a shooting test using two types of 9 mm projectiles (jacketed and unjacketed bullets). The ribs were then burnt until complete calcination in an electric furnace. The entry wound of each rib was analyzed using XRF, revealing traces of GSR. The XRF analysis showed that all samples, except for one, contain Pb and/or Sb near the lesion. Furthermore, the samples hit by unjacketed bullets had a more significant presence of Pb in macroscopic yellow areas, which persisted when moving away from the gunshot. These findings could pave the way for the use of XRF technology, mostly when a fast and immediate scan must be done on osteologic materials by a conservative method., (© 2024. The Author(s).)

Genetic and fragmented palaeoanthropological data suggest that Denisovans were once widely distributed across eastern Eurasia 1-3 . Despite limited archaeological evidence, this indicates that Denisovans were capable of adapting to a highly diverse range of environments. Here we integrate zooarchaeological and proteomic analyses of the late Middle to Late Pleistocene faunal assemblage from Baishiya Karst Cave on the Tibetan Plateau, where a Denisovan mandible and Denisovan sedimentary mitochondrial DNA were found 3,4 . Using zooarchaeology by mass spectrometry, we identify a new hominin rib specimen that dates to approximately 48-32 thousand years ago (layer 3). Shotgun proteomic analysis taxonomically assigns this specimen to the Denisovan lineage, extending their presence at Baishiya Karst Cave well into the Late Pleistocene. Throughout the stratigraphic sequence, the faunal assemblage is dominated by Caprinae, together with megaherbivores, carnivores, small mammals and birds. The high proportion of anthropogenic modifications on the bone surfaces suggests that Denisovans were the primary agent of faunal accumulation. The chaîne opératoire of carcass processing indicates that animal taxa were exploited for their meat, marrow and hides, while bone was also used as raw material for the production of tools. Our results shed light on the behaviour of Denisovans and their adaptations to the diverse and fluctuating environments of the late Middle and Late Pleistocene of eastern Eurasia., (© 2024. The Author(s).)

Rib spalling is one of the main factors restricting the safe and efficient production of the fully mechanized mining face in gangue-bearing coal seams, and the gangue has significant influence on the occurrence of rib spalling. In this study, the instability process and mechanical mechanism of rib spalling in gangue-bearing coal seams were studied, and the sensitivity of gangue parameters to rib spalling was analyzed. The simulation test of rib spalling under different gangue parameters was carried out by orthogonal tests. The width and depth of rib spalling were taken as evaluation indexes, and the influence of gangue parameters on the rib spalling was analyzed by variance analysis and significance tests. The results show that the failure process of rib spalling is characterized by the fracturing failure of the lower coal body, shear failure of the gangue layer, and the falling off of the upper coal body caused by the gravity; the gangue parameters (thickness, density, joint inclination, and internal friction angle) have an important influence on the sliding instability of the coal wall. In the sensitivity analysis, the influence of gangue parameters on the width of rib spalling is ordered as gangue density > joint inclination > gangue thickness > internal friction angle; the influence of gangue parameters on the depth of rib spalling is ordered as gangue density > joint inclination > internal friction angle > gangue thickness. Besides, the greater the gangue density, the less damage caused by stress concentration, and the lower the risk of rib spalling., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Cadaveric rib is used as a cartilage source for reconstructive rhinoplasty in patients who lack sufficient native septal cartilage; however, these grafts are known to warp. Objective: To measure and compare the biochemical properties of cadaveric rib as related to age, gender, and cortical versus core location. Methods: Seven cadaveric rib cartilage specimens were obtained and sectioned into cortical and core segments. Biochemical assays were used to determine total collagen and sulfated glycosaminoglycan (sGAG) content. Results: Collagen was present in higher amounts in cortical segments than core samples (72.8 ± 35.14 vs. 37.3 ± 16.99 μg/mgww, p  = 0.0005). sGAG was also shown to be more prevalent in cortical segments (25.47 ± 11.59 vs. 12.17 ± 7.15 μg/mgww, p  < 0.0001). The concentrations of collagen and sGAG demonstrated a positive correlation ( R 2  = 0.44, p  = 0.0004). Collagen and sGAG content decreased with the age of the donor ( p  = 0.001 and p  < 0.0001, respectively), but donor gender did not appear to affect collagen or sGAG content ( p  = 0.62 and p  = 0.43, respectively). Conclusion: Collagen and sGAG content was higher in cortical segments of cadaveric rib cartilage than in core segments, and higher in samples from younger cadavers as well.

Manufacturing of probiotics and functional foods using lactic acid bacteria (LAB) that overproduce vitamin B 2 has gained growing interest due to ariboflavinosis problems affecting populations of both developing and affluent countries. Two isogenic Lactiplantibacillus plantarum strains, namely a riboflavin-producing parental strain (UFG9) and a roseoflavin-resistant strain (B2) that carries a mutation in the FMN-aptamer of the potential rib operon riboswitch, were analysed for production and intra- and extracellular accumulation of flavins, as well as for regulation of the rib operon expression. Strain B2 accumulated in the medium one of the highest levels of riboflavin+FMN ever reported for LAB, exceeding by ~ 25 times those accumulated by UFG9. Inside the cells, concentration of FAD was similar in both strains, while that of riboflavin+FMN was ~ 8-fold higher in B2. Mutation B2 could decrease the stability of the aptamer's regulatory P1 helix even in the presence of the effector, thus promoting the antiterminator structure of the riboswitch ON state. Although the B2-mutant riboswitch showed an impaired regulatory activity, it retained partial functionality being still sensitive to the effector. The extraordinary capacity of strain B2 to produce riboflavin, together with its metabolic versatility and probiotic properties, can be exploited for manufacturing multifunctional foods., (© 2021 The Authors. Microbial Biotechnology published by Society for Applied Microbiology and John Wiley & Sons Ltd.)

The ANDE 6C Rapid DNA system could offer a potential alternative for the processing of calcified and soft tissue samples, often encountered in mass disaster scenarios. While originally designed for single source buccal swabs, interest in the performance of these instruments when using other types of single source samples continues to grow. To enhance the recovery of otherwise lesser quality samples, the manufacturer developed the investigative biochip, an alternative to the NDIS approved Arrestee biochip for reference sample buccal swabs. This study explores the viability of using the ANDE 6C system and the investigative biochip to process soft and calcified tissue, and uses conventional sample processing to contrast the results. Though the success rate obtained using the instrument's expert system was lower than expected - 0% muscle, 11% ribs, and 50% teeth -, the ANDE 6C offers an advantage over conventional calcified tissue processing in terms of turn-around time and processing complexity. If robust analysis parameters can be established to allow the evaluation of the generated data by a qualified analyst on a third party software platform, the use of the ANDE 6C and investigative biochip could be a suitable alternative for currently employed procedures. However, as is the case with conventional DNA typing, the quantity, age, type of biological material and quality of the exemplars could all play a role in the success of the ANDE 6C typing process. In addition, it appears as if the calcified tissue pre-processing protocol that provides the better opportunity for the ANDE 6C success is not appropriate to be carried out in the field or by non-trained personnel as special equipment as well as a certain level of exe expertise and technique is necessary. Nevertheless, disaster victim and unidentified human remain samples could be processed in a laboratory setting using the Rapid DNA ANDE 6C platform provided sufficient material is available to conduct a second, 'rescue' sample processing if necessary., (Copyright © 2021 Elsevier B.V. All rights reserved.)

While several studies have examined temporal DNA degradation in bones collected from terrestrial environments, studies on temporal DNA degradation in bones collected from aquatic environments are limited and mostly based on case studies. The objective of this study was to assess the impact of long-term submersion, aquatic environment, bone type and DNA extraction method on DNA quality and quantity. Bone samples (scapulae and ribs), collected every ~1000 ADD from a freshwater lake and river, underwent DNA extraction via ChargeSwitch ® gDNA Plant Kit and organic phenol-chloroform methods, and DNA quantitation using both TaqMan and SYBR Green-based quantitative PCR (qPCR) methods. Results suggest that in both bone types, quality of recovered DNA (i.e., degradation index) declined significantly with increase in submersion time. Among two bone types, quality of recovered DNA from scapulae declined faster than rib samples. There was no significant difference in recovered DNA quantity between bone types, DNA extraction methods, or locations but various interactions between these variables showed significant difference. Overall, it can be concluded that DNA can be extracted from waterlogged bone in sufficient quantity to generate an STR profile up to 4000 ADD., (© 2020 American Academy of Forensic Sciences.)

In forensic contexts of advanced decomposition, when conventional matrices are no longer available for toxicological analyses, finding alternative matrices is necessary. The skeleton, which is fundamental for anthropologists and geneticists, could be useful also for toxicological purposes. The present study aims to examine what kind of information toxicological analysis performed on bones (the cranium and the ribs) in different states of preservation could provide to the forensic practitioner. Thirty cadavers with known pharmacological history, subjected to forensic autopsy at the Institute of Legal Medicine of Milan, were selected. Rib and cranium samples were collected from each body and separated into two parts in order to create two different states of preservation: One was cleaned from soft tissues and analyzed as a well-preserved bone sample; the other was submitted to a long maceration process, simulating complete skeletonization. All specimens were then processed with accelerated solvent extraction and the eluates analyzed using Q-Exactive™ Orbitrap™ Mass Spectrometer. The analysis of blood and skeletal matrices showed positive results for the tested substances in 63% of cases, mainly benzodiazepines, antidepressants, and drug abuse. Significant Pearson correlations were observed between non-macerated vs. macerated bone samples: r = 0.79 for rib samples, r = 0.61 for cranium samples, and r = 0.69 for all bone samples. As a consequence, the positive results confirm the potential of the bone tissue as an alternative matrix in forensic toxicology, even in cases of extremely decomposed bodies. This study also highlighted important elements for reconstructing the biological profile in cases of forensic anthropological concern., (© 2020 American Academy of Forensic Sciences.)

Microenvironments play a significant part in understanding the post-mortem interval in forensic taphonomy. Recently, the value of weathering factors in relation to obtaining a PMI has been investigated further. In this study, observations were made to calculate the length of time it takes for three different bone elements (femur, rib, and scapula) to bleach in a UK summer and winter. This research also investigated whether there were any physicochemical modifications to the bone caused by bleaching. Porcine femora, scapulae, and ribs were placed into open and shaded areas of an outdoor research facility located in Oxfordshire, UK, during summer (July-Sep) and winter months (Dec-Mar). The specimens were monitored at 3-week intervals using photography, and an observational scoring method was developed to quantify the extent of bleaching. As temperatures are typically much lower in the UK compared with warmer climates, a controlled indoor-simulated desert experiment was also undertaken to be used as a control. This allowed sun bleaching and changes to the bone chemistry to be monitored in a controlled, high-UV environment for comparison with the UK outdoor experiments. Fourier transform infrared spectroscopy (FTIR) was employed to analyze physicochemical modifications to both the mineral and organic components of the bone. The FTIR was used to calculate crystallinity index (CI), mineral to organic ratio, and the relative amount of carbonate concentrations. Weather data was collected and a positive correlation was found between both ultraviolet (UV) levels and accumulated degree days (ADD) when compared with observational bleaching scores. Bleaching (whitening) of the bone samples occurred in both seasons but at different rates, with the bleaching process occurring at a slower rate in winter. During summer, the initial bleaching process was evident at 6 weeks, and by 9 weeks, the bones were an off-white colour. During the winter period, whitening of the bone started at 9 weeks; however, only the scapula and rib samples displayed a similar off-white colour. This colouration was observed at 13 weeks rather than at 9 weeks. The desert simulation samples started bleaching in a similar pattern to the outdoor samples after 1 week but the bones did not fully bleach. The bone chemistry, based on physicochemical properties obtained from the FTIR, showed a significant statistical difference between the simulated desert and winter season when compared against a control sample. For the winter samples, the mineral to organic ratio was significantly higher than that in the control, suggesting a reduction in the proportion of organic. For the samples in the simulated desert environment, the crystallinity index was significantly higher than that in the control samples, suggesting an increase in crystallinity. The results of this experiment support the fact that it is possible to achieve bleaching in a UK environment and that the minimal time frame for this to occur differs in seasons.

Most lizards walk and run with a sprawling gait in which the limbs are partly advanced by lateral undulation of the axial skeleton. Ribs and vertebrae are integral to this locomotor mode, but 3D motion of the axial skeleton has not been reported for lizard locomotion. Here, we use XROMM to quantify the relative motions of the vertebrae and ribs during slow treadmill locomotion in three savannah monitor lizards (Varanus exanthematicus) and three Argentine black and white tegus (Salvator merianae). To isolate locomotion, we selected strides with no concurrent lung ventilation. Rib rotations can be decomposed into bucket-handle rotation around a dorsoventral axis, pump-handle rotation around a mediolateral axis, and caliper rotations around a craniocaudal axis. During locomotion, every rib measured in both species rotated substantially around its costovertebral joint (8-17 degrees, summed across bucket, pump and caliper rotations). In all individuals from both species, the middle ribs rotated cranially through bucket and pump-handle motion during the propulsive phase of the ipsilateral forelimb. Axial kinematics during swing phase of the ipsilateral forelimb were mirror images of the propulsive phase. Although further work is needed to establish what causes these rib motions, active contraction of the hypaxial musculature may be at least partly responsible. Unilateral locomotor rib movements are remarkably similar to the bilateral pattern used for lung ventilation, suggesting a new hypothesis that rib motion during locomotion may have been an exaptation for the evolution of costal aspiration breathing in stem amniotes.

Objectives: The research explores whether the combined study of cortical bone histology, bone morphology, and dietary stable isotopes can expand insights into past human health and adaptations, particularly dietary sufficiency and life span., Materials and Methods: Midthoracic rib cortices from 54 South African Late Holocene adult skeletons (28 M, 24 F, two sex undetermined) are assessed by transmitted-light microscopy for cross-sectional area measurements, osteon area (On.Ar), osteon population density, and presence/absence of secondary osteon variants. Values for δ 13 C bone collagen , δ 15 N bone collagen , 14 C dates, Southwestern and Southern Cape geographic regions, body size measures, estimated ages-at-death from both morphological and histological methods are integrated into analyses, which include Spearman correlations, χ 2 tests and Kruskal-Wallis ANOVAs., Results: There is reduced On.Ar variability with higher δ 15 N (r = -.41, p = .005); rib %cortical area and δ 15 N are negatively correlated in the Southern Cape group (r = -.60, p = .03). Osteon variants are more common in older adults; histological ages at death are significantly older than those determined from gross morphology., Discussion: We found bone tissue relationships with measures of diet composition, but indicators of dietary adequacy remain elusive. Relationships of tissue quality and isotopes suggest that some Southern Cape adults lived long lives. Osteon variants are associated with age-at-death; some association with diet remains possible. Gross morphological methods appear to underestimate adult ages-at-death, at least among small-bodied adults., (© 2019 Wiley Periodicals, Inc.)

A method based on gas chromatography-mass spectrometry (GC-MS) is described for the determination of venlafaxine, amitriptyline and duloxetine in human bone. Pulverized samples were incubated in methanol for 1 h under ultrasonication, after the addition of sertraline as internal standard. The samples were centrifuged, and the supernatants were evaporated. Samples were then resuspended in 0.1 M phosphate buffer pH 6 and subjected to solid phase extraction. Chromatography was performed on a fused silica capillary column and analytes were determined in the selected-ion-monitoring (SIM) mode. The assay was validated in the range 0.3-1 ng/mg (depending on the drug) to 500 ng/mg. The mean absolute recoveries ranged from 92.6% to 96.2%, the matrix effect from 76.9% to 103.3% and process efficiency from 74% to 95.9% depending on the analyte. The intra- and inter-assay accuracy values were always better than 20%. The validated method was then successfully applied to real bone samples from forensic cases in which toxicological analysis for these drugs in blood had been positive. Drugs were detected in bone in all blood positive results, the approximate concentrations being 36.4 ng/mg for amitriptyline, 19.3-3 ng/mg for duloxetine and 4.6-2 ng/mg for venlafaxine., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study concentrates on samples of bare pork skin, with and without bristles, and dried bovine ribs shot with a semi-automatic pistol to find the best methodology and sampling surface in the search for inorganic gunshot residues (IGSR). Four quadrants of known surface areas were sampled at different distances from the bullet's hole with different swabs: tapes in graphite, Leukosilk® white tape, 3M® transparent tape, and a cotton swab to assess the technique able to collect the highest amounts of IGSR with the lowest contribution of the blank. The cotton swab wet in 10% HNO 3 gave the best results. The highest amounts of IGSR, measured by ICP-OES and MS, were detected on a surface of 3-cm radius from the bullet's edge. The amount of metals collected decreased with the firing distance between 20 and 60 cm. The procedure was efficient for sampling different tissues like skin and bones., (© 2018 American Academy of Forensic Sciences.)

Rib bone biopsy samples are often used to estimate changes in skeletal mineral reserves in cattle but differences in sampling procedures and the bone measurements reported often make interpretation and comparisons among experiments difficult. 'Full-core' rib bone biopsy samples, which included the external cortical bone, internal cortical bone and trabecular bone (CBext, CBint and Trab, respectively), were obtained from cattle known to be in phosphorus (P) adequate (Padeq) or severely P-deficient (Pdefic) status. Experiments 1 and 2 examined growing steers and Experiment 3 mature breeder cows. The thickness of cortical bone, specific gravity (SG), and the amount and concentration of ash and P per unit fresh bone volume, differed among CBext, CBint and Trab bone. P concentration (mg/cc) was closely correlated with both SG and ash concentrations (pooled data, r=0.99). Thickness of external cortical bone (CBText) was correlated with full-core P concentration (FC-Pconc) (pooled data, r=0.87). However, an index, the amount of P in CBext per unit surface area of CBext (PSACB; mg P/mm2), was more closely correlated with the FC-Pconc (pooled data, FC-Pconc=37.0+146×PSACB; n=42, r=0.94, RSD=7.7). Results for measured or estimated FC-Pconc in 10 published studies with cattle in various physiological states and expected to be Padeq or in various degrees of Pdefic status were collated and the ranges of FC-Pconc indicative of P adequacy and P deficiency for various classes of cattle were evaluated. FC-Pconc was generally in the range 130 to 170 and 100 to 120 mg/cc fresh bone in Padeq mature cows and young growing cattle, respectively. In conclusion, the FC-Pconc could be estimated accurately from biopsy samples of CBext. This allows comparisons between studies where full-core or only CBext biopsy samples of rib bone have been obtained to estimate changes in the skeletal P status of cattle and facilitates evaluation of the P status of cattle.

When deaths occur in water, soft tissue decomposes after a temperature-dependent period, making DNA identification dependent on bone. This study examined the effects of water on bone DNA purity and quantity, and determined the best of three extraction methods for isolating DNA. The organic phenol-chloroform method consistently extracted DNA approximating the accepted 260/280 purity value (~1.8); ChargeSwitch® gDNA Plant Kit and DNeasy Blood and Tissue Kit produced fair and unacceptable values, respectively. The purity value for humerus and rib samples was consistent across accumulated degree days (ADD). Significant differences in quantification among extraction methods and between bone types were identified. Ribs and ChargeSwitch® gDNA Plant Kit samples produced the lowest mean Ct values of the bone types and the extraction methods, respectively. Therefore, this study proposes that magnetic bead technology extraction methods and ribs be considered when processing bones that have been submerged in water for any length of time., (© 2018 American Academy of Forensic Sciences.)

The question of cognitive complexity in early Homo sapiens in North Africa is intimately tied to the emergence of the Aterian culture (~145 ka). One of the diagnostic indicators of cognitive complexity is the presence of specialised bone tools, however significant uncertainty remains over the manufacture and use of these artefacts within the Aterian techno-complex. In this paper we report on a bone artefact from Aterian Middle Stone Age (MSA) deposits in Dar es-Soltan 1 cave on the Atlantic coast of Morocco. It comes from a layer that can be securely dated to ~90 ka. The typological characteristics of this tool, which suggest its manufacture and use as a bone knife, are comparatively similar to other bone artefacts from dated Aterian levels at the nearby site of El Mnasra and significantly different from any other African MSA bone technology. The new find from Dar es-Soltan 1 cave combined with those from El Mnasra suggest the development of a bone technology unique to the Aterian., Competing Interests: The authors have declared that no competing interests exist.

The postmortem interval (PMI) of skeletal remains is a crucial piece of information that can help establish the time dimension in criminal cases. Unfortunately, the accurate and reliable determination of PMI from bone continues to evade forensic investigators despite concerted efforts over the past decades to develop suitable qualitative and quantitative methods. A relatively new PMI method based on the analysis of citrate content of bone was developed by Schwarcz et al. The main objective of our research was to determine whether this work could be externally validated. Thirty-one bone samples were obtained from the Forensic Anthropology Center, University of Tennessee, Knoxville, and the Onondaga County Medical Examiner's Office. Results from analyzing samples with PMI greater than 2 years suggest that the hypothetical relationship between the citrate content of bone and PMI is much weaker than reported. It was also observed that the average absolute error between the PMI value estimated using the equation proposed by Schwarcz et al. and the actual ("true") PMI of the sample was negative indicating an underestimation in PMI. These findings are identical to those reported by Kanz et al. Despite these results this method may still serve as a technique to sort ancient from more recent skeletal cases, after further, similar validation studies have been conducted., (© 2017 American Academy of Forensic Sciences.)

Bone tissue contains organic material that is useful for forensic investigations and may contain preserved endogenous protein that can persist in the environment for extended periods of time over a range of conditions. Single amino acid polymorphisms in these proteins reflect genetic information since they result from non-synonymous single nucleotide polymorphisms (SNPs) in DNA. Detection of genetically variant peptides (GVPs) - those peptides that contain amino acid polymorphisms - in digests of bone proteins allows for the corresponding SNP alleles to be inferred. Resulting genetic profiles can be used to calculate statistical measures of association between a bone sample and an individual. In this study proteomic analysis on rib cortical bone samples from 10 recently deceased individuals demonstrates this concept. A straight-forward acidic demineralization protocol yielded proteins that were digested with trypsin. Tryptic digests were analyzed by liquid chromatography mass spectrometry. A total of 1736 different proteins were identified across all resulting datasets. On average, individual samples contained 454±121 (x¯±σ) proteins. Thirty-five genetically variant peptides were identified from 15 observed proteins. Overall, 134 SNP inferences were made based on proteomically detected GVPs, which were confirmed by sequencing of subject DNA. Inferred individual SNP genetic profiles ranged in random match probability (RMP) from 1/6 to 1/42,472 when calculated with European population frequencies in the 1000 Genomes Project, Phase 3. Similarly, RMPs based on African population frequencies were calculated for each SNP genetic profile and likelihood ratios (LR) were obtained by dividing each European RMP by the corresponding African RMP. Resulting LR values ranged from 1.4 to 825 with a median value of 16. GVP markers offer a basis for the identification of compromised skeletal remains independent of the presence of DNA template., (Published by Elsevier B.V.)

This study investigates the use of Scanning electron microscopy-energy-dispersive X-ray (SEM-EDX) as a diagnostic tool for the determination of the osseous origin of samples subjected to different temperatures. Sheep (Ovis aries) ribs of two experimental groups (fleshed and defleshed) were burned at temperatures of between 100°C and 1100°C in 100°C increments and subsequently analyzed with the SEM-EDX to determine the atomic percentage of present elements. Three-factor ANOVA analysis showed that neither the exposure temperature, nor whether the burning occurred with or without soft tissue present had any significant influence on the bone's overall elemental makeup (p > 0.05). The Ca/P ratio remained in the osseous typical range of between 1.6 and 2.58 in all analyzed samples. This demonstrates that even faced with high temperatures, the overall gross elemental content and atomic percentage of elements in bone remain stable, creating a unique "fingerprint" for osseous material, even after exposure to extreme conditions., (© 2017 American Academy of Forensic Sciences.)

Fossilized organic remains are important sources of information because they provide a unique form of biological and evolutionary information, and have the long-term potential for genomic explorations. Here we report evidence of protein preservation in a terrestrial vertebrate found inside the vascular canals of a rib of a 195-million-year-old sauropodomorph dinosaur, where blood vessels and nerves would normally have been present in the living organism. The in situ synchrotron radiation-based Fourier transform infrared (SR-FTIR) spectra exhibit the characteristic infrared absorption bands for amide A and B, amide I, II and III of collagen. Aggregated haematite particles (α-Fe 2 O 3 ) about 6∼8 μm in diameter are also identified inside the vascular canals using confocal Raman microscopy, where the organic remains were preserved. We propose that these particles likely had a crucial role in the preservation of the proteins, and may be remnants partially contributed from haemoglobin and other iron-rich proteins from the original blood., Competing Interests: The authors declare no competing financial interests.

Citrate content in bone has been shown to be associated with the postmortem interval (PMI), with citrate decreasing after death as a function of time. Here we test this method using porcine ribs for the period of 1-165days after death, and also assess citrate content and variation from samples placed into two different postmortem environments (terrestrial and aquatic). Higher citrate variation, lower citrate recovery, and a weaker association with time were found in this study as compared to others. Citrate content, however, was found to decrease with increasing PMI, and the method was found to be easy and inexpensive to apply. No significant differences were found in citrate loss between terrestrial and aquatic environments. Although more research is needed, citrate content appears to be a promising new approach in estimating PMI from skeletal remains., (Published by Elsevier B.V.)

Objectives: A contribution to the knowledge of the economy and the environmental surroundings of the populations living along the Nile valley in three different periods., Materials and Methods: This study reports stable isotope analyses on apatite bone samples of 139 humans, 48 mammals, and 43 fish from the Al Khiday archaeological sites in Sudan. The bones belong to four archaeological periods: pre-Mesolithic, Mesolithic, Neolithic, and Meroitic. Data were processed statistically., Results: A significant difference exists between the pre-Mesolithic and Mesolithic mean δ(18) Ow value and the mean of the modern Nile. The mean δ(18) Ow values for the Neolithic humans and bovids are very similar (+1.5‰ ±4‰, and -2‰, respectively) and more positive than the mean values of Mesolithic mammals and Pre-Mesolithic humans. The water ingested by Meroitic humans (+7‰ ± 2.5‰) is enriched in (18) O in respect to the water ingested by the Neolithic population. There is a separation in the δ(13) Cdiet values between the pre-Mesolithic humans (-14‰ ± 1‰) and Mesolithic mammals (-12‰ ± 2‰) group and the Neolithic humans (-18‰ ± 1‰), Meroitic humans (-19‰ ±1‰), Neolithic mammals (-21‰), and the modern (mean δ(13) Cdiet  = -19‰ ±2‰) mammal group., Discussion: The climate became warmer and more arid from the pre-Mesolithic/Mesolithic to the Meroitic period. The environmental conditions influenced the strategies of subsistence and, in particular, the changes occurring from the pre-Mesolithic to the Neolithic can be considered contemporaneous to the transition from hunting-gathering-fishing to cultivation-herding. Am J Phys Anthropol 160:498-518, 2016. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

It is expensive and laborious to evaluate carcass composition in beef cattle. The objective of this study was to evaluate a method to predict the 9th to 11th rib section (rib) composition through empirical equations using dual energy X-ray absorptiometry (DXA). Dual energy X-ray absorptiometry is a validated method used to describe tissue composition in humans and other animals, but few studies have evaluated this technique in beef cattle, and especially in the Zebu genotype. A total of 116 rib were used to evaluate published prediction equations for rib composition and to develop new regression models using a cross-validation procedure. For the proposed models, 93 ribs were randomly selected to calculate the new regression equations, and 23 different ribs were randomly selected to validate the regressions. The rib from left carcasses were taken from Nellore and Nellore × Angus bulls from 3 different studies and scanned using DXA equipment (GE Healthcare, Madison, WI) in the Health Division at Universidade Federal de Viçosa (Viçosa, Brazil). The outputs of the DXA report were DXA lean (g), DXA fat free mass (g), DXA fat mass (g), and DXA bone mineral content (BMC; g). After being scanned, the rib were dissected, ground, and chemically analyzed for total ether extract (EE), CP, water, and ash content. The predictions of rib fat and protein from previous published equations were different ( < 0.01) from the observed composition. New equations were established through leave-one-out cross-validation using the REG procedure in SAS. The equations were as follows: lean (g) = 37.082 + 0.907× DXA lean ( = 0.95); fat free mass (g) = 103.224 + 0.869 × DXA fat free mass ( = 0.93); EE mass (g) = 122.404 + 1.119 × DXA fat mass ( = 0.86); and ash mass (g) = 18.722 + 1.016 × DXA BMC ( = 0.39). The equations were validated using Mayer's test, the concordance correlation coefficient, and the mean square error of prediction for decomposition. For both equations, Mayer's test indicated that if the intercept and the slope were equal to 0 and 1 ( > 0.05), respectively, then the equation correctly estimated the rib composition. Comparing observed and predicted values using the new equations, Mayer's test was not significant for lean mass ( = 0.26), fat free mass ( = 0.67), EE mass ( = 0.054), and ash mass ( = 0.14). We concluded that the rib composition of Nellore and Nellore × Angus bulls can be estimated from DXA using the proposed equations.

Objective: This study analyzes deciduous dental pathology and stable isotopes to investigate the relationship between diet, feeding practices, and oral health in a subadult skeletal sample from the Greek colonial site of Apollonia Pontica, Bulgaria (mid-5th to mid-3rd Centuries BC)., Methods: Stable carbon and nitrogen isotope analysis was conducted on 74 bone collagen samples, and the deciduous dentitions of 85 individuals aged 8.5 months to 11 years were examined for evidence of caries, calculus, antemortem tooth loss, abscesses, and occlusal tooth wear., Results: δ(13) C and δ(15) N values of the collagen samples indicate that weaning began between the ages of 6 months and 1 year, and was complete for most individuals by the age of 3. The isotopic data are consistent with a mixed diet of primarily terrestrial C3 resources. The dental pathology data indicate that complementary foods provided to young children had an impact on their oral health early on. Four outliers exhibited elevated δ(15) N values compared with the adult female range and lower levels of tooth wear than other members of their age groups. Possible explanations include prolonged breastfeeding, the consumption of diets elevated in (15) N, and physiological/nutritional stress., Conclusions: The deciduous dental data correlate well with the isotopic data and are consistent with later textual sources regarding the timing and duration of weaning, and the composition of complementary foods. The results of this research demonstrate the value of combining isotopic and dental evidence to investigate the dietary practices of infants and young children and the impact of these practices on oral health., (© 2015 Wiley Periodicals, Inc.)

Objective: The Okhotsk people were sedentary hunter-gatherer-fishers who lived and prospered in Sakhalin, Hokkaido, and the Kurile Islands during the 5th to 13th centuries AD. They expanded rapidly along the northeastern coast of Hokkaido. We reconstructed infant feeding practices of the Moyoro population of the Okhotsk culture in eastern Hokkaido, Japan., Methods: Stable isotope ratios in 58 subadult human skeletons were measured., Results: The results suggest that complementary foods with a relatively low carbon isotope ratio were consumed during and after weaning, as observed in ethnographic descriptions of northern human populations such as the Ainu and isotopically suggested in ancient northern hunter-gatherer-fisher populations. Nitrogen isotope ratios of subadults showed that the age at the end of weaning in the Moyoro population was 1.8 (1.4-2.2 in 95% credible interval) years, which is earlier than that in other northern hunter-gatherer-fisher populations., Conclusions: Because weaning age is one of the most important determinants of fertility, a shorter breastfeeding period suggests increased fertility. Furthermore, better nutrition would further promote the population increase, and thus populations of the Okhotsk culture could expand into new regions. These findings are consistent with recent emerging evidence of great contributions of the Okhotsk to the formation of later Ainu populations and culture., (© 2015 Wiley Periodicals, Inc.)

Little is known about the persistence of gunshot residue (GSR) in soft tissue and bones during decomposition in marine environments. For a better understanding, qualitative and quantitative data were obtained on GSR retention on soft tissue and bony gunshot wounds (GSWs). A quantity of 36 fleshed and 36 defleshed bovine ribs were shot at contact range with 0.22 calibre hollow point ammunition using a Stirling 0.22 calibre long rifle. Bone specimens in triplicate were placed in three environments: submerged, intertidal and in supralittoral zone. Sets of triplicates were recovered on day 3, 10, 24 and 38, and analysed with scanning electron microscopy with energy dispersive X-ray spectrometry (SEM-EDX), and inductive coupled plasma mass spectrometry (ICP-MS). The SEM-EDX recorded GSR-indicative particles surrounding the bullet entrance on all bone types (fleshed and defleshed) in all environments throughout the study. GSR-unique particles were only detected on the supralittoral bones. The ICP-MS analysis showed faster GSR loss on submerged than intertidal and supralittoral defleshed specimens. Fleshed specimens showed a faster GSR loss on intertidal than submerged and supralittoral specimens. In conclusion, the GSR disappeared faster from submerged and intertidal than non-submerged specimens. The difference of detection of GSR between analysed specimens (defleshed versus fleshed) disappeared upon defleshing. This study highlights the potential of finding evidence of GSR in a submerged body and the potential of microscopic and analytical methods for examining suspected GSW in highly decomposed bodies in marine habitats., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Here we report bone phosphate oxygen (δ(18)Op) values from perinates/neonates and infants (<3.5 years; n = 32); children (4-12 years; n = 12); unsexed juveniles (16-18 years; n = 2); and adult bones (n = 17) from Wharram Percy, North Yorkshire, England, in order to explore the potential of this method to investigate patterns of past breastfeeding and weaning. In prior studies, δ(15)N and δ(13)C analyses of bone collagen have been utilized to explore weaning age in this large and well-studied assemblage, rendering this material highly appropriate for the testing and development of this alternative method targeting the inorganic phase of bone. Data produced reveal (18)O-enrichment in the youngest perinatal/neonatal and infant samples, and an association between age and bone δ(18)Op (and previously-published δ(15)N values), with high values in both these isotope systems likely due to breastfeeding. After the age of 2-3 years, δ(18)Op values are lower, and all children between the ages of 4 and 12, along with the vast majority of sub-adults and adults sampled (aged 16 to >50 years), have δ(18)Op values consistent with the consumption of local modern drinking water. The implications of this study for the reconstruction of weaning practices in archaeological populations are discussed, including variations observed with bone δ(15)Ncoll and δ(18)Op co-analysis and the influence of culturally-modified drinking water and seasonality. The use of this method to explore human mobility and palaeoclimatic conditions are also discussed with reference to the data presented., (© 2015 Wiley Periodicals, Inc.)

A multi-isotope investigation (Sr and Pb isotopes and δ18O, δ13C and δ15N) was applied to bone and teeth from an unidentified male found drowned in the"IJ" Ruyterkade in Amsterdam, The Netherlands in March of 1999. The individual remained unidentified until mid 2013, after the isotope study was completed. Coupled δ13C and δ15N values in bone collagen recovered from rib and femur are consistent with an omnivore living in a region where C3-type diet dominates (i.e. Europe). Integrated Sr and Pb isotopes and δ18O values in canine and third molar teeth and femur and rib bone data exclude extended residence in north-west Europe and particularly The Netherlands. Characteristic Pb isotope ratios coupled with inferred δ18O values of drinking water argue for a most probable place of origin for the unidentified individual in west and south Poland, south-east Slovakia and the region of Ukraine-Romania-Bulgaria, specifically the region associated with the Carpathian Mountains. Independent of the isotope study, the Cold Case Team made a positive identification with an individual from south-west Poland, validating the results of the multiple-isotopic approach., (Copyright © 2014 Forensic Science Society. Published by Elsevier Ireland Ltd. All rights reserved.)

While recent forensic research has focused on determining which skeletal elements are superior in their preservation of DNA over the long term, little focus has been placed on measuring intra-element variation. Moreover, there is a general belief that dense (cortical) bone material will contain better-preserved DNA than does spongy (cancellous) bone. To address these ideas, quantitative PCR was used to estimate the degree of mitochondrial DNA (mtDNA) preservation variance across sections of 19 northern fur seal ribs (Callorhinus ursinus) that date to ∼3000 years before present. Further, we developed a measure called the "density index" that was used to gauge the relative densities of the rib sections studied here to determine if density was an appropriate predictor of preservation. The average preservation among the samples was significantly different (ANOVA, p=1.9×10(-9)) with only 15% of the total variance observed within samples. However, 12 of the 19 specimens (∼63.2%) exhibited at least an order of magnitude difference in mtDNA preservation across the whole. Regression of the amount of mtDNA extracted per gram of bone material against the density index of the bone from which it was extracted demonstrates no relationship between these variables (R(2)=0.03, p=0.28)., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

The challenge of depth-resolved, nonionizing (hybrid-optical) detection of mineral loss in bones is addressed using truncated-correlation photothermal coherence tomography (TC-PCT). This approach has importance not only in ground-based clinical procedures, but also in microgravity space applications. Analogous to x-ray morphometric parameters, two- and three-dimensional markers have been defined and estimated for chemically demineralized goat rib bones. Cortical and trabecular regions have been analyzed independently and together using the computational slicing advantage of TC-PCT, and the results have been verified using micro-CT imaging (the gold standard). For low-demineralization levels, both modalities follow the same trend. However, for very high mineral loss that is unlikely to occur naturally, anomalies exist in both methods. Demineralization tracking has been carried out to a depth of ∼3  mm below the irradiated surface. Compared with micro-CT imaging, TC-PCT offers an improved dynamic range, which is a beneficial feature while analyzing highly demineralized bones. Also, TC-PCT parameters are found to be more sensitive to trabecular and combined cortical-trabecular demineralization compared with x-ray parameters. Axial and lateral resolutions in bone imaging for the current instrumental configuration are ∼25 and 100 μm, respectively.

Objective: Microtia is treated with rib cartilage sculpting and staged procedures; though aesthetically pleasing, these constructs lack native ear flexibility. Tissue-engineered (TE) elastic cartilage may bridge this gap; however, TE cartilage implants lead to hypertrophic changes with calcification and loss of flexibility. Retaining flexibility in TE cartilage must focus on increased elastin, maintained collagen II, decreased collagen X, with prevention of calcification. This study compares biochemical properties of human cartilage to TE cartilage from umbilical cord mesenchymal stem cells (UCMSCs). Our goal is to establish a baseline for clinically useful TE cartilage., Methods: Discarded cartilage from conchal bowl, microtic ears, preauricular tags, rib, and TE cartilage were evaluated for collagen I, II, X, calcium, glycosaminoglycans, elastin, and fibrillin I and III. Human UCMSCs were chondroinduced on 2D surfaces and 3D D,L-lactide-co-glycolic acid (PLGA) fibers., Results: Cartilage samples demonstrated similar staining for collagens I, II, and X, elastin, and fibrillin I and III, but differed from rib. TE pellets and PLGA-supported cartilage were similar to auricular samples in elastin and fibrillin I staining. TE samples were exclusively stained for fibrillin III. Only microtic samples demonstrated calcium staining., Conclusions: TE cartilage expressed similar levels of elastin, fibrillin I, and collagens I and X when compared to native cartilage. Microtic cartilage demonstrated elevated calcium, suggesting this abnormal tissue may not be a viable cell source for TE cartilage. TE cartilage appears to recapitulate the embryonic development of fibrillin III, which is not expressed in adult tissue, possibly providing a strategy to control TE elastic cartilage phenotype.

Rib collagen of 51 juveniles and 11 adult females from the late medieval Fishergate House cemetery site (York, UK) were analyzed using nitrogen and carbon stable isotope ratio analysis to determine the weaning age for this population and to reconstruct diet. The juveniles' ages ranged from fetal to 5-6 years, while the females were of reproductive age. Previous researchers suggested that the children from Fishergate House might have been weaned later than the medieval British norm of 2 years, based on a mortality peak at 4-6 years of age. The results show weaning was complete by 2 years of age, agreeing with previous British weaning studies. The adult female δ(15) N values have a mean of 11.4‰ ± 1.1‰ and the δ(13) C values have a mean of -19.4‰ ± 0.4‰. These findings are consistent with previous isotopic studies of female diet in York during this period, though slightly lower. The weaned juvenile nitrogen values were found to be higher than the adult females (12.4‰ ± 1.0‰ for δ(15) N and -19.7‰ ± 0.5‰ for δ(13) C), which might indicate a dependence on higher trophic level proteins such as marine fish or pork. Marine fish is considered a high status food and children are considered low-status individuals at this time, making this a particularly interesting finding. Weaning does not appear to coincide with peak mortality, suggesting environment factors may be playing a larger role in child mortality at Fishergate House., (Copyright © 2013 Wiley Periodicals, Inc.)

Studies of low copy number (LCN) and degraded DNA are prone to contamination from exogenous DNA sources that in some cases out-compete endogenous DNA in PCR amplification, thus leading to false positives and/or aberrant results. Particularly problematic is contamination that is inadvertently deposited on the surfaces of bones through direct handling. Whereas some previous studies have shown that contamination removal is possible by subjecting samples to sodium hypochlorite prior to DNA extraction, others caution that such treatment can destroy a majority of the molecules endogenous to the sample. To further explore this topic, we experimentally contaminated ancient northern fur seal (Callorhinus ursinus) ribs with human DNA and treated them with sodium hypochlorite to remove that contamination. Our findings are consistent with previous studies that found sodium hypochlorite to be highly efficient (~81-99%) at contamination removal; however, there emerged no treatment capable of removing 100% of the contamination across all of the experiments. Moreover, the ability to estimate the degree of damage to endogenous northern fur seal molecules was compromised due to the inherent variability of preserved mtDNA across the bones, and the presence of co-extracted PCR inhibitors., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

This study tests the hypothesis that vertical habitat preferences of different monkey species inhabiting closed canopy rainforest are reflected in oxygen isotopes. We sampled bone from seven sympatric cercopithecid species in the Taï forest, Côte d'Ivoire, where long-term study has established taxon-specific patterns of habitat use and diet. Modern rib samples (n = 34) were examined for oxygen (δ(18)Oap) and carbon (δ(13)Cap) from bone apatite ('bioapatite'), and carbon (δ(13)Cco) and nitrogen (δ(15)Nco) from bone collagen. Results are consistent for C3 feeders in a closed canopy habitat. Low irradiance and evapotranspiration, coupled with high relative humidity and recycled CO2 in forest understory, contribute to observed isotopic variability. Both δ(13)Cco and δ(13)Cap results reflect diet; however, δ(13)C values are not correlated with species preference for canopy height. By contrast, δ(18)Oap results are correlated with mean observed height and show significant vertical partitioning between taxa feeding at ground, lower and upper canopy levels. This oxygen isotope canopy effect has important palaeobiological implications for reconstructing vertical partitioning among sympatric primates and other species in tropical forests.

Gunshot residue (GSR) has been sought and demonstrated on many types of material and with many techniques. Inductively coupled plasma optical emission spectrometry (ICP-OES) could be a useful method on difficult substrates, but a systematic study on burnt material has never been performed. Hence, this study aims at evaluating the usefulness and reliability of this method on burnt samples. Sixteen adult bovine ribs (eight with soft tissues, eight totally skeletonized) were shot using two kinds of projectile (both 9 mm full metal-jacketed or unjacketed). Then, every sample was led to complete calcination in an electric oven. The area of the gunshot entrance wound was swabbed and analyzed by ICP-OES; the results were also correlated with a previously published parallel study by scanning electron microscopy (SEM) equipped with an SEM-energy dispersive X-ray analyzer. ICP-OES proved to be very sensitive and reliable even on degraded material and can be an appropriate nondestructive method for detecting residues on difficult and delicate substrates such as burnt bone., (© 2013 American Academy of Forensic Sciences.)

This study was undertaken to provide reference values of chemical element mass fractions in intact bone of Reference (European Caucasian) Man/Woman. The rib bone samples investigated were obtained from autopsies of 84 apparently healthy 15-58-year-old citizens (38 females and 46 males) of a non-industrial region in the Central European part of Russia who had suffered sudden death. The mass fractions (mg/kg given on a wet mass basis) of 69 elements in these bone samples were measured by using neutron activation analysis with high-resolution spectrometry of short-lived and long-lived radionuclides, particle-induced gamma-ray emission, inductively coupled plasma atomic emission spectrometry, and inductively coupled plasma mass spectrometry including necessary quality control measures. Using published and measured data, mass fraction values of the 79 elements for the rib bone have been derived. Based on accepted rib to skeleton mass fractions and reference values of skeleton mass for Reference Man, the elemental burdens in the skeleton were estimated. These results may provide a representative bases for establishing related reference values for the Russian Reference Man/Woman and for revising and adding current reference values for the International Commission on Radiological Protection. The data presented will also be very valuable for many other applications in radiation protection, radiotherapy radiation dosimetry, and other scientific fields.

The medieval period in Europe was a time of unprecedented social complexity that affected human diet. The diets of certain subgroups-for example, children, women, and the poor-are chronically underrepresented in historical sources from the medieval period. To better understand diet and the distribution of foods during the medieval period, we investigated stable carbon and nitrogen isotope ratios of 30 individuals from Trino Vercellese, Northern Italy (8th-13th c.). Specifically, we examined diet differences between subgroups (males and females, and high- and low-status individuals), and diet change throughout the life course among these groups by comparing dentine and bone collagen. Our results show a diet based on terrestrial resources with input from C(4) plants, which could include proso and/or foxtail millet. Diets of low-status males differ from those of females (both status groups) and of high-status males. These differences develop in adulthood. Childhood diets are similar among the subgroups, but sex- and status-based differences appear in adulthood. We discuss the possibility of cultural buffering and dietary selectivity of females and high-status individuals., (Copyright © 2012 Wiley Periodicals, Inc.)

The research and analysis of gunshot residues has a relevant role in the examination of gunshot wounds. Nevertheless, very little literature exists concerning gunshot wounds on charred material. In this study, 16 adult bovine ribs (eight still with soft tissues and eight totally skeletonized) underwent a shooting test with two types of projectiles (9 mm full metal-jacketed bullet and 9 mm unjacketed bullet). Each rib then underwent a charring process in an electric oven, reaching the stage of complete calcination at 800°C. The area of each entrance wound was analyzed before and after the carbonization process via a scanning electron microscope (SEM) equipped with an energy dispersive X-ray analyzer (EDX). In each sample, metallic residues composed of lead, barium, and antimony were found. These metallic residues were thus preserved also after exposure to the extremely high temperatures reached within the oven, especially with unjacketed bullets, although the particles seem to be more irregular in shape as a result of the heating process. In conclusion, this study proved that gunshot residues survive extremely high temperatures and can be detected via SEM/EDX even in cases of charred tissues.

In literature, many studies have been performed in order to investigate the presence of GSR ("gunshot residue") and metallic residues in general with radiological techniques on several types of material, but the survival of metallic residues on charred samples has never been systematically performed. In this study, 31 adult bovine ribs underwent a shooting test. Every rib was shot with a single bullet, at a near-contact shooting distance, using two kinds of projectile: 17 samples were shot with a full metal-jacketed bullet and the remaining 14 with an unjacketed bullet. After the shooting test, every rib underwent a "charring cycle" in an electric oven up to 800°C. Every sample underwent radiological investigation with conventional radiography, before and after the burning process, to evaluate any changes in number and distribution of metallic residues. Radiographs showed survival of radiopaque residues in every sample, even after the charring process, especially when the bullet used was of the unjacketed type.

We present stable isotopic analyses of collagen from 80 servicemen excavated from the late 18th/early 19th century naval hospitals at Plymouth (50) and Haslar, Gosport (30) in southern England. Historical records suggest that, the diets of these two populations should be essentially identical. While δ(15) N of the rib collagen confirmed that naval servicemen were relatively well-catered for in terms of meat allowance (Plymouth average δ(15) N = 11.1‰, Gosport = 11.9‰), stable carbon isotope analysis produced average values for the two assemblages, which were significantly different (Plymouth average δ(13) C = -18.8‰, Gosport = -20.0‰). We postulate that these differences stem from divergent naval postings, with a greater proportion of Plymouth individuals serving in areas that entailed a greater input of C(4) foodstuffs. By comparison with published data from approximately contemporary burials at Snake Hill, Ontario, Canada and Chesapeake Bay, Virginia, we suggest that this area is the east coast of North America. For 15 of the 30 individuals from Gosport, we have data on ribs, femur, and dentine from the same skeleton, which appear to show that they came from a variety of locations in their preadolescence, but converged in dietary terms onto a "naval average," which is consistent with historical evidence for recruitment patterns into the Navy at the time. By comparison with published data from skeletons recovered from the wreck of the Mary Rose (sank 1545), we conclude that this naval diet was virtually unchanged from the 16th century to the end of the 18th century., (Copyright © 2012 Wiley Periodicals, Inc.)

The goal of this research was to develop empirical equations to predict chemical and physical compositions of the carcass and the body using the composition of the 9th- to 11th-rib section (rib(9-11)) and other measurements. A database (n = 246) from 6 studies was developed and comprised 37 bulls (BU), 115 steers (STR), and 94 heifers (HF), of which 132 were Nellore (NEL), 76 were NEL × Angus crossbreds (NA), and 38 were NEL × Simmental crossbreds (NS). The right half carcass and the rib(9-11) from the left half carcass were analyzed for ether extract (EE), CP, and water. The remaining components were chemically analyzed to determine the composition of the body. A stepwise procedure was used to determine the variable inclusion in the regression models. The variables included were EE in the rib(9-11) (EER; %), CP in the rib(9-11) (CPR; %), water in the rib(9-11) (WR; %), visceral fat (VF; %; KPH and mesenteric fats), organs plus viscera (OV; %), carcass dressing percentage (CD; %), cold carcass weight (kg), and empty BW (EBW; kg). No sex or breed effects were found on EE and CP compositions of the carcass (C(EE) and C(CP), respectively; %); the equations were as follows: C(EE) = 4.31 + 0.31 × EER + 1.37 × VF [n = 241; R(2) = 0.83; mean square error (MSE) = 4.53] and C(CP) = 17.92 + 0.60 × CPR - 0.17 × CD (n = 238; R(2) = 0.50; MSE = 1.58). Breed affected water content in the carcass (C(W), %); the equations were as follows: C(W) = 48.74 + 0.28 × WR - 0.017 × EBW for NEL; C(W) = 46.69 + 0.32 × WR - 0.017 × EBW for NA; and C(W) = 38.06 + 0.48 × WR - 0.017 × EBW for NS (n = 243; R(2) = 0.67; MSE = 5.17). A sex effect was found on body chemical EE composition (BW(EE)); the equations were as follows: BW(EE) = 2.75 + 0.33 × EER + 1.80 × VF for BU; BW(EE) = 1.84 + 0.33 × EER + 1.91 × VF for STR; and BW(EE) = 4.77 + 0.33 × EER + 1.28 × VF for HF (n = 243; R(2) = 0.89; MSE = 3.88). No sex or breed effects were found on CP composition in the body (BW(CP)); the equation was as follows: BW(CP) = 14.38 + 0.24 × CPR (n = 240; R(2) = 0.59; MSE = 1.06). A sex effect was found for body water content (BW(W)); the equations were as follows: BW(W) = 38.31 + 0.33 × WR - 1.09 × VF + 0.50 × OV for BU; BW(W) = 45.67 + 0.25 × WR - 1.89 × VF + 0.50 × OV for STR; and BW(W) = 31.61 + 0.47 × WR - 1.06 × VF + 0.50 × OV for HF (n = 241; R(2) = 0.81; MSE = 3.84). The physical carcass composition indicated a breed effect on all components and a sex effect for fat in the carcass. We conclude that body and carcass compositions can be estimated with rib(9-11) for purebred and crossbred NEL animals, but specific equations have to be developed for different groups of animals.

The presented study deals with the effect of the cremation temperature on the microstructure and morphology of the human compact bone. The biological material consisted of samples from ribs of recent Central European origin belonging to individuals of known age, sex and cause of death. Each bone sample was divided into several sections. One section remained unburned and the rest were burned at 700, 800 and 1000 degrees C. A few samples were burned also at the temperature of 600 degrees C. The undecalcified unstained ground cross-sections were made from burned and unburned bones; photographed and analysed using the SigmaScan Pro 5 programme. During burning, both the macroscopic and microscopic dimensions of the bone shrink, including the measures of the individual microstructures. The percentual representation of the area of individual microstructures on the area of the cross-section decreases. The number of individual microstructures per mm2 of the compact bone cross-section increases. Most microstructural variables demonstrated statistically significant differences at the individual temperatures of cremation. The burned bones showed a large scale of the colours, especially at 700 degrees C.

GJA1 gene (Connexin43, also known as Cx43) is the most abundant gap junction protein isoform in animal cells and is associated with bone development in embryos. The objective of the present work was to evaluate in vivo osteal development in GJA1-deficient fetal mice through determination of the histological and molecular alterations induced by partial or total deletion of the GJA1 gene. Heterozygous C57BL/6 mice (HT) harboring a null mutation of the GJA1 gene were mated, and pregnant females were submitted to euthanasia and Caesarean section from 12.5 to 19.5 days post coitum (dpc). HT (GJA1(+/-)) and homozygous (GJA1(-/- )) knockout (KO) mutants and wild-type (WT) fetuses were identified by polymerase chain reaction (PCR), and development curves were constructed on the basis of fetus weight and crown-rump length. Histopathological, histochemical, and real-time PCR analyses were performed in order to assess the expression of markers associated with bone development, namely, osteocalcin, osteopontin, alkaline phosphatase, RUNX2, GJA1, GJC1 (Cx45), and GJA3 (Cx46). HT and KO fetuses exhibited delays in the differentiation of osteoblasts and, consequently, in bone development in comparison with the WT group. Additionally, less deposition of mineralized and osteoid matrix was observed in GJA1-deficient fetuses. Bone development in KO fetuses was delayed through the moment of birth, but in HT animals the delay only extended until 17.5 dpc, following which development was normalized. The expression of genes coding for osteocalcin, osteopontin, alkaline phosphatise, and RUNX2 were also delayed in GJA1-deficient fetuses. Animals that exhibited a lower expression of GJA1 presented delayed expression of the GJC1 and GJA3 genes and their corresponding protein products in the bone tissue. The results of the present study contribute to our understanding of the function of GJA1 during bone development and suggest that GJC1 could play a role in restoring intercellular communication in GJA1-deficient mice.

Purpose: Brachytherapy planning software relies on the Task Group report 43 dosimetry formalism. This formalism, based on a water approximation, neglects various heterogeneous materials present during treatment. Various studies have suggested that these heterogeneities should be taken into account to improve the treatment quality. The present study sought to demonstrate the feasibility of incorporating Monte Carlo (MC) dosimetry within an inverse planning algorithm to improve the dose conformity and increase the treatment quality., Methods and Materials: The method was based on precalculated dose kernels in full patient geometries, representing the dose distribution of a brachytherapy source at a single dwell position using MC simulations and the Geant4 toolkit. These dose kernels are used by the inverse planning by simulated annealing tool to produce a fast MC-based plan. A test was performed for an interstitial brachytherapy breast treatment using two different high-dose-rate brachytherapy sources: the microSelectron iridium-192 source and the electronic brachytherapy source Axxent operating at 50 kVp., Results: A research version of the inverse planning by simulated annealing algorithm was combined with MC to provide a method to fully account for the heterogeneities in dose optimization, using the MC method. The effect of the water approximation was found to depend on photon energy, with greater dose attenuation for the lower energies of the Axxent source compared with iridium-192. For the latter, an underdosage of 5.1% for the dose received by 90% of the clinical target volume was found., Conclusion: A new method to optimize afterloading brachytherapy plans that uses MC dosimetric information was developed. Including computed tomography-based information in MC dosimetry in the inverse planning process was shown to take into account the full range of scatter and heterogeneity conditions. This led to significant dose differences compared with the Task Group report 43 approach for the Axxent source., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Breastfeeding patterns were subject to a number of fads in 18th and 19th century Britain. Feeding infants by hand, rather than maternal breastfeeding or wet-nursing, became more prevalent among both the wealthy and poor. Substitute foods may have been a convenient alternative for mothers employed away from the household. This study used stable isotope ratio analysis to examine the weaning schedule in the 18th and 19th century skeletal assemblage from Spitalfields, London, UK. Analysis of 72 juvenile ribs revealed δ(15) N elevations of 2-3‰ above the adult mean for individuals up to the age of two, while elevations of 1-2‰ were observed in δ(13) C for the first year of life. This suggests that the introduction of solid foods took place before the end of the first year, and that breastfeeding had entirely ceased by 2 years of age. The age at death of many of these infants is known from historical records, and can be used to pinpoint the amount of time required for the breast milk signal to be observed in the stable isotope ratios of rib collagen. Results show that a δ(15) N elevation can be detected in the ribs of individuals who died as young as 5-6 weeks. Not all individuals at Spitalfields were breastfed, and there may not have been a single uniformly practiced weaning scheme. There is, however, more evidence for prolonged breastfeeding during the 19th century than the 18th century., (2011 Wiley Periodicals, Inc.)

For the first time, the contents of rare earth elements (REEs) in a rib bone of a healthy human were determined. The mean value of the contents of Ce, Dy, Er, Gd, La, Nd, Pr, Sm, Tb, and Yb (10 elements out of 17 total REEs), as well as the upper limit of means for Ho, Lu, Tm, and Y (4 elements) were measured in the rib bone tissue of 38 females and 42 males (15 to 55 years old) using inductively coupled plasma mass spectrometry (ICP-MS). We found age-related accumulation of REEs in the bone tissue of healthy individuals who lived in a non-industrial region. It was calculated that during a lifespan the content of REEs in a skeleton of non-industrial region residents may increase by one to two orders of magnitude. Using our results as indicative normal values and published data we estimated relative Gd accumulation in the bone tissue of patients according to magnetic resonance imaging with contrast agent and La accumulation in the bone tissue of patients receiving hemodialysis after treatment with lanthanum carbonate as a phosphate binder. It was shown that after such procedures contents of Gd and La in the bone tissue of patients are two to three orders of magnitude higher than normal levels. In our opinion, REEs incorporation may affect bone quality and health similar to other potentially toxic trace metals. The impact of elevated REEs content on bone physiology, biochemistry and morphology requires further investigation.

We measured stable nitrogen isotope ratios in bone collagen of 60 individuals from the early Neolithic (9th-8th millennium Cal. BC) sites of Çayönü Tepesi and Aşıklı Höyük. Our aim was to identify the duration of exclusive breastfeeding (EBF), compare this with juvenile mortality at each site, and assess whether there was a relationship between them. The isotope analysis suggests that weaning commenced at about 1 year at Aşıklı Höyük and around 2 years at Çayönü Tepesi. The mortality data show equal numbers of infant deaths up to 24 months; however, after 24 months, the mortality rate increases at Çayönü Tepesi, and a Student's t-test confirms a significant difference in infant mortality between the sites. Weaning foods prepared in the early Neolithic from agricultural crops would have had low-iron content, poor nutritional value, and would have been prepared in nonsterilized containers. Therefore, later weaned infants in early Neolithic farming settlements, although capable of some immunological response, were probably undernourished putting them at a disadvantage when encountering bacteria in their weaning food. Our results suggest that infant feeding regimes that introduced infants to weaning foods in the first year of life may have had a positive effect on their survival., (© 2010 Wiley-Liss, Inc.)

Skeletal tissues from a domestic pig exposed to amitriptyline, diazepam, and pentobarbital were analyzed to determine the relative distribution of these drugs in bone. Following drug exposure and euthanasia, remains were allowed to decompose outdoors to complete skeletonization between summer 2007 and fall 2009. Remains were recovered and separated according to bone type. Twelve different bone types were pulverized and sampled in triplicate. Each bone sample underwent methanolic extraction (96 h, 50 °C), followed by solid-phase extraction and gas chromatography-mass spectrometry in the selected ion monitoring mode. Mass-normalized assay responses underwent ANOVA with post-hoc testing, revealing bone type as a main effect for all three drugs, but not for the diazepam metabolite (nordiazepam). The assay response varied with respect to bone type by factors of 27, 39, and 20 for pentobarbital, diazepam, and amitriptyline, respectively. The relative distribution between bone type was qualitatively similar for the three administered drugs analyzed for, with the largest response obtained from rib for all three drugs. This is the first study, to the authors' knowledge, of the distribution of different drugs in various decomposed skeletal tissues in a controlled experiment using an animal model of comparable physiology to humans. These data have implications for the interpretive value of forensic drug measurements in skeletal tissues.

Pork ribs with intact muscle tissue were used in an experimental attempt to identify bullet wipe on bone at distances from 1 to 6 feet with 0.45 caliber, full metal jacket ammunition. This resulted in the unexpected finding of primer-derived gunshot residue (GSR) deep within the wound tract. Of significance is the fact that the GSR was deposited on the bone, under the periosteum, after the bullet passed through a Ziploc bag and c. 1 inch of muscle tissue. It is also important to note that the GSR persisted on the bone after the periosteum was forcibly removed. The presence of primer-derived GSR on bone provides the potential to differentiate gunshot trauma from blunt trauma when the bone presents an atypical gunshot wound. In this study, the presence of gunshot primer residue at a distance of 6 feet demonstrates the potential for establishing maximum gun-to-target distance for remote shootings.