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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).)

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.

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.)

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.

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.)

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.

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.)

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.

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.)

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 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.

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.

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.

Tuberculosis has been a scourge of humans over many millennia, but questions remain regarding its evolution and epidemiology. Fossil biomarkers, such as DNA and long-chain mycolic acids, can be detected in ancient skeletal and other materials. The phthiocerol dimycocerosate waxes are also robust biomarkers for tuberculosis and sensitive methods are available for the detection of their mycocerosic acid components. The presence of mycocerosic acids was investigated in 49 individuals from the 1837-1936 Coimbra Identified Skeletal Collection (Portugal), half with documentary data indicating tuberculosis as a cause of death. Samples were hydrolysed, acidic components converted to pentafluorobenzyl esters, the non-hydroxylated long-chain esters isolated, and this fraction separated into multimethyl-branched and other esters by normal phase high performance liquid chromatography. Negative ion chemical ionisation gas chromatography mass spectrometry was used to detect diagnostic C29, C30 and C32 mycocerosic acids. Mycocerosic acids were detected in archaeological material for the first time, illustrating that they are valuable biomarkers for the diagnosis of ancient tuberculosis. A 72% correlation with the Coimbra burial record supported TB as the major cause of death. In addition, 30% of the skeletons, positive for mycocerosates, showed the presence of related long-chain mycolipenic acids.

The effect of age and gender on major, minor, and trace element contents in the intact rib bone of 80 relatively healthy 15-55-year-old women and men was investigated. Contents or upper limit of contents of 16 chemical elements in the rib bone were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES). Mean values (M +/- SEpsilonMu) for the mass fraction of Ba, Ca, Cu, Fe, K, Li, Mg, Na, P, S, Sr, and Zn (milligram per kilogram of dry bone) were as follows: 2.54 +/- 0.16, 171,400 +/- 4,050, 1.35 +/- 0.22, 140 +/- 11, 1,874 +/- 71, 0.049 +/- 0.011, 2,139 +/- 38, 5,378 +/- 88, 75,140 +/- 1,660, 1,881 +/- 51, 291 +/- 20, and 92.8 +/- 1.5, respectively. The upper limits of contents of Al, B, Mn, and V were <7.20, <0.65, <0.36, and <0.03, respectively. Statistically significant tendency for the Ca, Mg, and P content to decrease with age was found in the human rib bone, regardless of gender. The mass fraction of Fe in the male rib bone increases with age. It was shown that higher Ca, Mg, Na, P, and Sr mass fractions as well as lower Fe content were typical of female ribs as compared to those in male ribs.

Several lineages of teleost fishes have independently derived skeletons composed solely of acellular bone, a tissue without obvious advantages over bone that has osteocytes in the matrix. There is no consensus for the functional role of acellular bone, as factors such as salinity, activity level and gross morphology have been shown to be poor predictors of acellularity. We used a three-point bending method to test the hypothesis that the material stiffness (elastic modulus) of acellular bone is higher than that of cellular bone, which could be evidence that material properties were a selective pressure in the evolution of this unusual skeletal material. The acellular ribs of Myoxocephalus polyacanthocephalus are curved, hollow beams that decrease in size both distally and posteriorly along the rib series. First and second moments of area decreased distally and caudally in all individuals. Young's modulus (E) ranged from 3.67 to 8.40 GPa, with a mean of 6.48 GPa. The flexural stiffness (EI) differed significantly between ribs, and the hollow cylinder morphology increased the flexural stiffness by 12.0% over a solid, circular cross-section rod with the same area. Contrary to our expectations, acellular bone is not stiffer by virtue of fewer lacunae but instead falls at the very low end of the range of stiffness seen in cellular bone. There remains the possibility that other properties (e.g. fatigue resistance, toughness) are higher in acellular bone.

The accumulation of natural and artificial radionuclides in humans and domestic animals is of interest in estimating effective doses of exposed humans and to decide whether animal products can be used for nutrition of the population. In this paper we present an investigation of the (137)Cs- and (40)K-activity levels of the ribs and sternum of a "mountain pasture" cow, born in a highly contaminated region of Styria, Austria, at the time of the radioactive fallout following the Chernobyl accident. This is the first systematic investigation of the variation in activity levels of a contaminated animal. The International Commission on Radiological Protection (ICRP) assumes that cesium and potassium are homogenously distributed throughout the whole body of an organism. However, the presented results show that there is a non-uniform distribution of (137)Cs and (40)K in different skeletal bones and their adherent tissues of a dairy cattle. We found that activity concentrations of (137)Cs and (40)K varied up to a factor 2.5 in different components of the ribs. The minimum values of (137)Cs and (40)K in the ribs were 29.9 and 21Bqkg(-1) fresh mass for trabecular bone in the vertebral half of asternal ribs, and the maximum values 332 and 132Bqkg(-1) fresh mass for a mixed sample composed of a cartilaginous tissue layer and parts of the perichondrium, both originating from asternal costal cartilages, respectively.

Although much literature is available on DNA extraction from tissue samples to obtain the best possible genotyping results, to the best of our knowledge no written recommendations exist on how to excise or extract bone and tooth samples from a victim to facilitate this. Because the possibility of cross-contamination is high, especially when excising numerous samples under disaster conditions, it is important to minimize this risk and to keep samples in optimum condition. In this paper a standard operating procedure is proposed for collection of femur, rib, and tooth samples to aid victim identification both after mass disasters and in (single) forensic investigations.

We analyzed lead concentrations in bones from both genders of Japanese merchants (including rohnin; masterless samurai) and farmer classes, and compared the findings with those of the samurai class in the Edo period (1603-1867) to clarify gender and hierarchical (or occupational) differences in lead exposure during the Japanese feudal age. Merchant class females had significantly higher lead exposure (90.8 microg Pb/g dry bone; n=20) than males of the same class (39.9 microg Pb/g dry bone; n=31) (p<0.01), indicating a remarkable gender difference in the urban population. In contrast to these high concentrations, males and females of the farmer class living in agricultural (or semi-rural) areas had significantly lower exposure (total mean value; 9.2 mug Pb/g dry bone; n=4) than both genders of the merchant class (p<0.001), and the gender difference was not significant in this class.

The extraction of DNA from archaeological or forensic skeletal remains can provide quite powerful data for analysis, but is plagued by a unique set of methodological problems. One of the most important methodological problems to overcome in such analyses is the presence of modern contamination on the surfaces of bones and teeth, which can lead to false positives and erroneous results unless it is removed before DNA extraction is initiated. Ancient DNA (aDNA) researchers and forensic scientists have employed a number of techniques to minimize such contamination. One such technique is the use of bleach (sodium hypochlorite--NaOCl) to "destroy" contaminating DNA. However, a consensus on the optimum concentration of sodium hypochlorite to be used and the amount of time the bone or tooth should be exposed to it has not emerged. The present study systematically approaches the issue by introducing contamination to ancient bones (from approximately 500 BP) and determining which of several sodium hypochlorite treatments best eliminates surface contamination. The elimination of surface contamination from bone requires immersion in at least 3.0% (w/v) sodium hypochlorite (approximately equal parts of commercial bleach and water) for at least 15 min. Endogenous DNA proved to be quite stable to even extreme sodium hypochlorite treatments (6% for 21 h), suggesting that DNA adsorbs to hydroxyapatite in the bone and that this process facilitates the preservation of DNA in ancient skeletal remains.

Desmoplastic fibroma is a very rare primary tumor of bone, closely related to aggressive fibromatosis of soft tissue. Although considered a benign lesion, it can be very aggressive locally and has a high rate of local recurrence after incomplete surgical excision. Radiologically, the lesion shows no distinctive features, often simulating osteosarcoma, fibrous dysplasia, or fibrosarcoma. Rib involvement by desmoplastic fibroma is extremely rare. We present two cases of desmoplastic fibroma involving this unusual location with lytic costal lesion and chest wall extension. Histological examination after surgical resection revealed that the tumor consisted of spindle cells with small, elongated nuclei in a background of numerous collagen fibers and infiltrating lamellar bone. There was no evidence of nuclear atypia, mitotic activity, or necrosis. We also discuss histological differential diagnosis as well as clinical features and the radiological and pathologic findings of this rare disease. Recognition of this entity is important to ensure proper surgical treatment.

The neuropeptide galanin (GAL) has recognized physiological actions in the nervous system and other tissues, but there is no documented evidence of GAL influencing normal or pathological bone metabolism. GAL expression, however, is upregulated in central and peripheral nerves following axotomy and is known to influence neural regeneration. Thus, severance of skeletal-associated nerves during fracture could similarly increase local GAL concentrations and thereby influence fracture healing. The initial aim of this study was therefore to identify the presence of GAL in normal bone and/or fracture callus by assessing the concentration and cellular localization of GAL in intact and/or fractured rat rib, using radioimmunoassay and immunohistochemistry, respectively. Groups of Sprague-Dawley rats (13 weeks old) had their left sixth ribs surgically fractured or underwent sham surgery and then calluses and nonfractured rib samples were analyzed at 1 and 2 weeks postsurgery (n = 5-6 per group). Low (basal) concentrations of GAL were detected in control ribs, whereas at 1 and 2 weeks postfracture, callus samples contained markedly increased levels of peptide ( approximately 32- and 18-fold increase, respectively, relative to controls; P < 0.01), revealing a strong upregulation during bone healing. Plasma GAL concentrations were also increased at 2 weeks postfracture (P < 0.005). In normal (nonfractured) rib, minimal levels of GAL-like immunoreactivity (LI) were present in cortical bone, periosteum, endosteum, and surrounding skeletal muscle. In costal cartilage plates, intense GAL-LI was present in all chondrocytes of the hypertrophic zone and in a population of chondrocytes in the reserve zone. GAL-LI was not present, however, in chondrocytes in the proliferative zone of costal cartilage or skeletal muscle fibers. In fracture callus, levels of GAL-LI were moderate to intense in osteoprogenitor cells and osteoblasts, in some chondrocytes, and in cartilaginous, osseous, and periosteal matrices. Subsequent studies revealed the presence of galanin receptor-1-like immunoreactivity (GALR1-LI) in most cell types shown to contain GAL-LI, although the distribution of GALR1-LI was more extensive in reserve zone chondrocytes than that of GAL-LI; and GALR1-LI also appeared in late proliferative zone chondrocytes of costal cartilage. In summary, GAL concentrations were significantly increased in fracture callus and plasma of rats that underwent rib fracture. In addition, GAL- and GALR1-LI was also detected in specific cells and structures within costal cartilage, bone, and fracture callus. These results strongly implicate GAL in aspects of cartilage growth plate physiology and fracture repair, possibly acting in an autocrine/paracrine fashion via GALR1.

Calcium and phosphorus concentrations as well as the Ca/P ratio were estimated in intact rib bone samples from healthy humans, 37 women and 45 men, aged 15-55 yr. For Ca and P concentration measurements, instrumental neutron activation analysis was used. The mean values (mean +/- SD) for the investigated parameters were 19.3 +/- 4.5% of dry bone weight, 8.42 +/- 2.14% of dry bone weight, and a ratio of 2.33 +/- 0.34, respectively. Statistically significant differences for the above parameters were not observed to be related either to age or sex. The mean values for Ca, P, and the Ca/P ratio were within a very wide range of published data and close to their medians. The individual variation for the Ca/P ratio in rib bone from healthy humans was lower than those for Ca and P taken separately. An indication is that the specificity of the Ca/P ratio improves upon that for Ca and P concentrations and may be more reliable in the diagnosis of bone disorders.

This study has directed attention at the search for bone-related proteins in an extract of demineralized rib bone of the 120 mya Iguanodon. The inner compact bone was demineralized and the GuCl extract resolved into 11 fractions using anion exchange chromatography, which all contained silver-reactive proteins with various amino acid profiles. Two specific fractions, iv and xi, revealed characteristics typical of contemporary phosphoproteins and proteoglycans, respectively. Fraction iv, 43-57 kDa, contained a high ratio of aspartate and serine, although no phosphate was discernable. Fraction xi contained a band of 41-47 kDa and was rich in chondroitin sulphate and hyaluronan. In addition an early eluting fraction was immunoreactive with an antibody against osteocalcin. A cancellous bone fraction from the same bone sample was also analyzed using N-terminal sequencing and revealed potential similarities with cystatin. While we do not claim to have identified the presence of intact proteins, this study has value in demonstrating that extruded extracellular matrix is protected by its capacity to induce mineralization, which subsequently is important in conserving detectable protein products in ancient skeletal tissues.

The Ca/P ratio was estimated in intact rib bone samples from healthy humans, 37 women and 45 men, aged from 15 to 55 years using instrumental neutron activation analysis. No statistically significant differences (p>0.05) age- or sex-related differences in the Ca/P ratio were observed. The mean value (M+/-SD) for the investigated parameter for the whole group studied, 2.33+/-0.34, was within a very wide range of published data and close to the median value.

We report on the measurements of carbon and nitrogen stable isotopes of both bone and teeth from a single site and population (Medieval Wharram Percy), undertaken to explore variations due to weaning in a past population. There have been a number of recent studies of weaning using delta(15)N values of ribs, and we indicate a number of assumptions that must be met before the results of such studies can be correctly interpreted. We found that rib collagen delta(15)N values decrease to adult levels after age 2 years, indicating that weaning occurred at or before this age. Rib collagen delta(13)C values are also more enriched than adult delta(13)C values before age 2 years, and we argue that this is due to the so-called "carnivore" effect in delta(13)C. We measured teeth and rib delta(15)N values from the same individuals and found that for individuals up to age 11 years, tooth dentine delta(15)N is higher than adult rib delta(15)N values, indicating that the dentine was formed during breast-feeding and that there was almost no turnover of dentine since. We observed some decrease in delta(13)C and delta(15)N rib values, compared to adult rib and teeth values, for the few years after weaning that may relate to a theoretically predicted physiological nitrogen imbalance during this period of rapid growth, but this is more likely due to a childhood diet (up to age 9) which was isotopically different from later diet, possibly consisting of a greater proportion of plant foods., (Copyright 2002 Wiley-Liss, Inc.)

A new method for the determination of ultra-trace level of lanthanides such as La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm Yb and Lu in human rib is explored. Closed-vessel microwave digestion with concentrated HNO3 is used for the sample preparation and the digested analyte is directly analyzed by ICP-MS. The method is sensitive and accurate and, is efficient for the determination of the fourteen lanthanides listed above. Detection limit for various elements ranges from 0.0007 microgram/L to 0.005 microgram/L. Calibration plots exhibit good linearity with high linear correlation coefficient r > or = 0.997. Recoveries of spiked samples ranges from 93.7% to 115.4%. Precision is < 3% RSD. Rhodium is used as the internal standard to compensate the signal suppression caused by the sample matrix. Isobaric overlap of polyatomic ions caused by matrix oxide/hydroxide of barium is corrected by interference correction factor. Two standard reference materials, namely "Human Hair GBW07601" and "Wheat Flour GBW08503" are used for quality control of the analyzed samples.

Osteocalcin is one of the most abundant noncollagenous proteins in bone. It is strongly associated with the mineral phase of bone, and has long been associated as a marker of bone turnover. However, its relationship to bone composition, strength, and structure is unclear. Carp rib bone is an excellent model for the study, because osteocalcin represents almost 60% of the total extractable noncollagenous proteins found in it. Because of the abundance of osteocalcin relative to other extractable proteins, any changes in the properties of carp rib bone would be more likely influenced by the osteocalcin concentration. To test the hypotheses that the concentration of osteocalcin is reflected in other properties of bone, the correlations between the osteocalcin concentration and the mineral content, microstructural properties, and physical characteristics of the bone mineral crystals were determined utilizing radioimmunoassay (RIA), spectrophotometry, nanoindentation, and small-angle X-ray scattering (SAXS) techniques, respectively. Osteocalcin concentration was found to be correlated to the molar Ca/P ratio and inversely correlated to the elastic modulus and hardness in the longitudinal plane. This study provides evidence for a putative relationship between the concentration of osteocalcin and the microstructural mechanical properties of bone. Correlations were also found between the mechanical properties in the longitudinal plane and both the phosphate content and the molar Ca/P ratio. However, no relationships could be identified between osteocalcin concentration and several parameters of bone crystals, as determined by SAXS.

The instrumental neutron activation analysis method was used for the determination of trace elements in rib bone samples taken from autopsies of accident victims. The elements Br, Ca, Cl, Cr, Fe, Mg, Mn, Na, P, Sr, Rb, and Zn were determined in cortical tissues by using short and long irradiations with thermal neutron flux of the IEA-R1m nuclear reactor. The reference materials NIST SRM 1400 Bone Ash and NIST SRM 1486 Bone Meal were also analyzed in order to evaluate the precision and the accuracy of the results. It was verified that lyophilization is the most convenient process for drying bone samples because it does not cause any element losses. Comparisons were made between the results obtained for rib samples and the literature values as well as between the results obtained for different ribs from a single individual and for bones from different individuals.

The use of the polymerase chain reaction (PCR) to define deoxyribonucleic acid (DNA) types at several loci was investigated. PCR was used to amplify nine short tandem repeat (STR) loci along with the amelogenin locus on the X and Y chromosomes using the AmpF/STR Profiler Plus PCR amplification kit (Perkin Elmer). Rib bones were collected from 12 individuals. Five cm portions were buried at a depth of approximately 30 cm and 5 cm portions were left on the surface of the ground. Samples were exposed to the environment for periods of time ranging from two weeks to 17 months. Dried blood standards were prepared for use as reference standards for each rib sample. Bone, muscle, and bone marrow were collected from each sample. DNA from each tissue type was extracted. Complete profile results were obtained from the surface bone samples out to an exposure time of 17 months. None of the muscle or bone marrow samples produced complete profile results beyond eight weeks. All DNA typing results from complete or incomplete profiles were consistent with DNA typing results of the corresponding blood standard. Results suggest that using the AmpF/STR Profiler Plus PCR Amplification Kit is a valid way to establish the DNA profile of tissue types from human remains.

We detected nerve growth factor (NGF) by immunohistochemical localization in both fractured and unfractured rat rib. In unfractured bone, periosteal mesenchymal osteoprogenitor cells appeared to be the only skeletal cells which stained for NGF. Adjacent skeletal muscle fibers exhibited NGF staining both in fractured and unfractured bone. Fracture callus periosteal osteoprogenitor cells, marrow stromal cells, osteoblasts, young osteocytes and endothelial cells of new capillaries had moderate to heavy staining for NGF at 1 and 3 weeks after fracture. Deeply positioned osteocytes and osteoclasts showed no NGF staining. Most chondrocytes of fracture calluses stained for NGF, however, some chondrocytes did not stain which may indicate that NGF is produced at particular stages of chondrocytic differentiation. In calluses, periosteal matrix stained heavily for NGF when juxtaposed to cartilage and less obviously when associated with new bone at both 1 and 3 weeks post-fracture. However, other fibrous, cartilaginous and osseous matrices did not stain for NGF at any time. At 6 weeks post-fracture, NGF staining was largely confined to periosteal osteoprogenitor cells. The detection of NGF in periosteal osteoprogenitor cells of unfractured rib points to these cells having a role in nerve maintenance in intact bone. Furthermore, the localization of NGF in osteoprogenitor cells, marrow stromal cells, osteoblasts, certain chondrocytes, endothelial cells, periosteal matrix of the fracture callus and skeletal muscle may mean that these entities participate in fracture innervation. The presence of NGF in the callus may also indicate a direct, as yet undefined action of this neurotrophin on skeletal cell metabolism.

Ceramiclike xenogeneic bone (CXB) was obtained from the fresh bone of pig ribs being treated by physical and chemical methods to deprive of its organic substance. The CXB possessed the same natural porous network system as that of the human. The CXB was cultured with the bone marrow stromal cells of rabit. When the marrow cells had integrated with the CXB, thus a new material was obtained. (CXB-BM), and was implanted sacro-spinal muscle of rabbit. The specimens were observed under phase microscope, light microscope and electronic scanning microscope. The results showed that: at the 2nd week after the implantation of CBX-BM composite material there began the new bone formation, and the rate of bone formation was increased with time. There was evident new bone formation after 24 weeks. The process of the new bone formation were quite similar to the composite graft of HAP red autogenous and marrow, but the former degraded faster and formed typical cancellous structure earlier. There was no new bone formation when CXB was implanted alone in the control. Both the mechanism of osteogenetic potential and its clinical application were discussed.

A rare variant of dedifferentiated chondrosarcoma with malignant mesenchymomatous component in a 57-year-old male is reported. The patient presented with a posterior mediastinal mass arising from the left eighth and ninth ribs showing well differentiated, low-grade chondrosarcoma. Five years later, local recurrence occurred and an excised specimen also showed the same histological features as the primary tumor. Another 6 years later, the tumor recurred and metastasized to the multiple organs, the patient dying 4 months later. Autopsy revealed that the recurrent and metastatic tumors showed malignant mesenchymomatous 'dedifferentiation' of chondrosarcoma composed of rhabdomyosarcoma, angiosarcoma, chondrosarcoma, osteosarcoma, and leiomyosarcoma, in addition to fibrosarcomatous areas. Although the less differentiated component of dedifferentiated chondrosarcoma usually shows the histological features of malignant fibrous histiocytoma and fibrosarcoma, multilineage differentiation can occur in that component. The phenomenon of 'dedifferentiation' in chondrosarcoma and the relationship to and distinction from malignant mesenchymoma of soft tissue and bone are discussed.

Element concentrations in ribs obtained from elderly Japanese people (17 males and 28 females; mean age, 81.5 years) were determined by atomic absorption spectrometry (AAS), inductively coupled plasma atomic emission spectrometry (ICP-AES), and ICP mass spectrometry (ICP-MS). Nine elements--Na, Mg, P, K, Ca, Fe, Zn, Sr, and Pb--were determinable in most of the subjects by a combination of AAS and ICP-AES. The levels of these elements were generally comparable with those obtained in our previous study on ribs from younger Japanese. By the use of ICP-MS, Sn (median, 0.79 micrograms/g dry bone) and Ba (1.3 micrograms/g) were determinable in all of the subjects analysed (n = 35) and 18 other elements at lower concentration levels were also detected in some of the subjects. An exploratory statistical analysis was carried out to find element(s) of which level(s) in rib vary in the presence of degenerative chronic diseases, using information obtained from pathological autopsy reports and medical histories of the present subjects. It indicated that (i) Pb and Zn, (ii) Ba, and (iii) Sr levels in the ribs varied in the presence of cancer, cerebrovascular damage, and bone problems, respectively. The present results were discussed in relation to the results of the previous epidemiologic and experimental studies.