High DNA yield from metatarsal and metacarpal bones from Slovenian Second World War skeletal remains.

DNA yield varies by anatomical region, and the selection of bone types that yield maximum recovery of DNA is important to maximize the success of human identification of skeletal remains. The goal of our study was to explore inter- and intra-individual variation in DNA content by measuring nuclear DNA quantity and quality and autosomal STR typing success to determine the most promising skeletal elements for bone sampling. To exclude the influence of taphonomic issues as much as possible, three complete male skeletons from a single Second World War mass grave were examined and all representative skeletal element types of the human body were analyzed. Forty-eight different types of bones from the head, torso, arm, leg, hand, and foot were sampled from each skeleton, 144 bones altogether. The samples were cleaned, and half a gram of bone powder was decalcified using a full demineralization extraction method. The DNA was purified in a Biorobot EZ1 (Qiagen). DNA content and rates of DNA degradation were determined with the PowerQuant (Promega), and the Investigator ESSplex SE QS (Qiagen) was used for STR typing. The highest-yielding bones mostly produced the most complete STR profiles. Among the skeletal elements containing on average the most DNA and producing the most complete profiles in all three skeletons examined were metacarpals, metatarsals, and the petrous portion of the temporal bone. Metatarsals and metacarpals can easily be sampled without using a saw, thus reducing potential DNA contamination. Skeletons from the Second World War can be used as a model for poorly preserved skeletal remains, and the results of the investigation can be applied for genetic identification of highly degraded skeletal remains in routine forensic casework. Although the research was limited to only three skeletons found in a unique mass grave, the data obtained could contribute to sampling strategies for identifying old skeletal remains. More Second World War skeletons will be analyzed in the future to investigate inter-bone variation in the preservation of DNA.

Immediate, short and medium-term effects of orthopedic insoles with a metatarsal retro-capital bar on biomechanical variables, plantar pressures and muscle activity in running.

BACKGROUND: The aim of this study was to evaluate the effect of 12 weeks of use of orthopedic insoles equipped with a metatarsal retro-capital bar (MRCB) on plantar pressure under the feet and lower limb kinematic variables during running. METHODS: Two groups of 10 runners used for 12 weeks while running orthopedic insoles without correction or equipped with a MRCB. All participants performed successively a standing posture (CoP displacement) test and a running test at 11 km.h-1 (lower limb kinematic variables) using with flat insoles and orthopedic neutral or MRCB insoles at the beginning (T0), after 4 (T4) and 12 weeks (T12) of use. RESULTS: For the MRCB group, CoP moved backwards while forefoot plantar pressure was decreased during standing position at T4 and T12 compared to T0. During running, the plantar pressure under the 2nd, 3rd and 4th metatarsal heads was reduced with MRCB at T0, T4 and T12. The one under the 1st metatarsal head was decreased at T4 and T12, when MRCB or flat insoles were used. The maximal extension and the total amplitude of ankle were slightly increased at T4 and T12 with or without wearing MRCB insoles. Similar changes in knee joint kinematics were observed but only at T12. Any significant changes were found in runners that used orthopedic insoles without correction. CONCLUSIONS: Orthopedic insoles equipped with MRCB involve lower plantar pressure under the metatarsal heads, which may be of interest to treat forefoot injuries in runners.

Citrate Content of Bone as a Measure of Postmortem Interval: An External Validation Study.

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.

Medieval cattle from Bern (Switzerland): an archaeozoological, genetic and historical approach.

This study deals with genetic analyses of an assemblage of mediaeval (13th century) cattle metapodials from Bern that had previously been osteometrically examined regarding sex, shape and wither height. The results from the genetic sexing of these small (height 100 to 120 cm) cattle correlate well with the osteometric interpretations. Some few exceptions we interpreted as cows used as draft animals with stouter bones and thus osteometrically determined as males. Two morphologically different groups of cow metatarsals however, we took as proof of the historical fact that Bern relied on livestock from different geographical origins: the town's vicinity and the alpine pastures with their favourable grazing conditions. It was not possible to distinguish them genetically. An analysis of one single nucleotide polymorphism (SNP) in the melanocortin receptor 1 (MC1R) showed that predominant coat colour most likely was red-brown. Furthermore, an analysis of the SNP in the Y-chromosomal intron UTY19 that divide modern taurine cattle in two major haplogroups (Y1 and Y2) showed that the mediaeval cattle belonged to the haplogroup Y2 with one single exception of a Y1.

Histological characterization of the early stages of bone morphogenetic protein-induced osteogenesis.

On the basis of currently available knowledge, we hypothesize that the initial bone formation, as induced by bone morphogenetic protein (BMP), is influenced by the chemical composition and three-dimensional spatial configuration of the used carrier material. Therefore, in the current study, the osteoinductive properties of porous titanium (Ti) fiber mesh with a calcium phosphate (Ca-P) coating (Ti-CaP), insoluble bone matrix (IBM), fibrous glass membrane (FGM), and porous particles of hydroxy apatite (PPHAP) loaded with rhBMP-2 were compared in a rat ectopic assay model at short implantation periods. Twelve Ti-CaP, 12 IBM, 12 FGM, and 12 PPHAP implants, loaded with rhBMP-2, were subcutaneously placed in 16 Wistar King rats. The rats were sacrificed at 3, 5, 7, and 9 days post-operative, and the implants were retrieved. Histological analysis demonstrated that IBM and Ti-CaP had induced ectopic cartilage and bone formation by 5 and 7 days, respectively. However, in PPHAP, bone formation and cartilage formation were seen together at 7 days. At 9 days, in Ti-CaP, IBM, and PPHAP, cartilage was seen together with trabecular bone. At 9 days, in FGM, only cartilage was observed. Quantitative rating of the tissue response, using a scoring system, demonstrated that the observed differences were statistically significant (Wilcoxon rank sum test, p < 0.05). We conclude that IBM, CaP-coated Ti mesh, FGM, and PPHAP provided with rhBMP-2 can indeed induce ectopic bone formation with a cartilaginous phase in a rat model at short implantation periods. Considering the different chemical composition and three-dimensional spatial configuration of the carrier materials used, these findings even suggest that endochondral ossification is present in rhBMP-2-induced osteogenesis, even though the amount of cartilage may differ.

Mechanical properties and chemical composition of avian long bones.

We have studied the mechanical behaviour of avian long bones as whole structures, by calculating mechanical parameters such as maximum load, stiffness, bending strength and flexural Young's modulus; bones were always tested in three-point bending. Furthermore composition in several chemical elements and amino acids related to collagen content was also analysed. Correlations were established between body mass, mechanical parameters and chemical contents. Both bending strength and Young's modulus were negatively correlated to body mass. Significant correlations were found between nitrogen content and both strength and Young's modulus, with negative slopes in both cases. Magnesium and phosphorus appear to be the most important inorganic elements to the understanding of the mechanical behaviour of avian long bones.

Osteocyte-bone lining cell system at the origin of steady ionic current in damaged amphibian bone.

A wound-generated steady electric current was measured by a two-dimensional vibrating probe system in the metatarsal bones of 22 adult frogs (Xenopus laevis) placed in amphibian Ringer. Inward currents were recorded entering a micrometric hole drilled through the cortex at middiaphyseal level. These steady state currents (mean +/- SD 8.50 +/- 2.77 microA/cm2) last approximately 2 hours, were dependent on the presence of sodium in the incubation medium, were no more detectable after fixation, and were reduced to background level when the cell membranes were solubilized. These results agree with previous recordings of metatarsal bones of weanling mice, under identical conditions. Both results suggest that the measured ionic currents have a cellular origin. Metatarsal bones of adult amphibian were purposely selected for this study because, unlike mammalian bones, their shafts are avascular and only contain an osteocyte-bone lining cell system, as documented by scanning and transmission electron observations. Thus, unlike the data from previous investigations on mammals, the results succeeded in giving the first convincing evidence that the osteocyte-bone lining cell system is the origin of damage-generated ionic currents. As damage exposes bone ionic compartment to plasma, damage-generated ionic currents are representative of ion fluxes at bone plasma interface, and cells at the origin of the current generate the driving force of such fluxes. By demonstrating that osteocytes and bone lining cells are at the origin of the current, this study suggests that the osteocyte-bone lining cell system, though operating as a cellular membrane partition, regulates ionic flow between bone and plasma. Since strain-related adaptive remodeling could also depend on ionic characteristics and flow of the bone fluid through the osteocyte lacuno-canalicular network, the results reported here support the view that osteocyte and bone lining cells may constitute a functional syncytium involved in mineral homeostasis as well as in bone adaptation to mechanical loading.

Fluoride exposure in cervids inhabiting areas adjacent to aluminum smelters in Norway. II. Fluorosis.

Mandibles from 1104 red deer (Cervus elaphus), 147 moose (Alces alces), and 453 roe deer (Capreolus capreolus), collected between 1990 and 1993 in the vicinity of seven Norwegian aluminum smelters, were examined for dental fluorotic and osteofluorotic lesions. The metacarpal or metatarsal bones from 214 of these cervids also were evaluated. Dental fluorotic lesions occurred in all three cervid species. Prevalence of dental fluorosis was generally low at the various locations, with the exception of Ardal, where 15% of the cervids examined were affected. Only sporadic cases of severe dental fluorotic lesions were diagnosed. All red deer yearlings (1.5 yr) with mandibular fluorine (F) levels exceeding 2,000 ppm F, had dental fluorosis. However, the lowest skeletal fluorine level found in a fluorotic animal of this age was 1,355 ppm F. Gross osteofluorosis occurred in only three cervids, all with mandibular fluorine residues > 8,000 ppm F. Hence, generalized fluorosis was not a prominent feature in the material studied.

The proto-oncogene c-myc is involved in cell differentiation as well as cell proliferation: studies on growth plate chondrocytes in situ.

A combination of immunocytochemistry and microdensitometry has been used to localize and quantify the expression of the proto-oncogene c-myc within chondrocytes of the proximal growth plates of rat and chick long bones. Although the c-myc protein was localized in all chondrocytes of the growth plate of both species the most intense staining was restricted to the proliferating and differentiating chondrocytes. These were identified by their ability to synthesize DNA (bromodeoxyuridine positive) and the presence of alkaline phosphatase activity, respectively. Species differences did exist with the c-myc concentration of the chick proliferating and differentiating chondrocytes being higher (128% and 240%, respectively) than the respective chondrocytes of the rat. The higher c-myc concentration in the chick proliferating chondrocytes paralleled the differences in the bromodeoxyuridine labelling index between the two species. In the rat, the concentration of c-myc protein present in the differentiating chondrocytes was 74% higher than in the respective proliferating chondrocytes, while in the chick it was 146% higher. The data not only provides further evidence for a role of the c-myc protein in cell proliferation but also suggests involvement of this protein in chondrocyte differentiation and/or hypertrophy.

Evaluation of a bone biopsy technique for determining the calcium and phosphorus status of swine from weaning to market weight.

Tibia biopsies were taken from 75 live pigs at 4-wk intervals and from 251 slaughtered pigs to evaluate bone biopsy as a procedure for determining Ca/P status in pigs fed 70, 85, 100, 115 and 130% of the NRC (1979) estimated dietary Ca and P percentage requirements from weaning to market. Least squares means and SE of live and slaughter biopsy wet weight, ash weight and dry, fat-free ash percentage (DFF%) were compared at each time in each trial and found not to differ. Diet and time effects on ash weight, ash percentage of wet weight and DFF% of the biopsy core also did not differ greatly between slaughter and live biopsies and generally responded linearly and quadratically (P less than .01) to increasing Ca/P level and time. Biopsy measures were correlated (P less than .05) with third and fourth metacarpal and metatarsal length, bending and shear stress and DFF%. Means for slaughter biopsy DFF% did not differ greatly from the average of third and fourth metacarpal and metatarsal DFF% from slaughter pigs. Means for live and slaughter biopsy DFF% were lower than those for whole bones for the 70 and 85% NRC estimated Ca/P levels, but not for the 100, 115 and 130% NRC levels. Bone biopsy offers potential as a reliable noninvasive procedure for monitoring Ca/P status of swine from weaning to market, but it needs further study for use in Ca/P research in swine.

Deposition of amyloid of unknown origin in articular cartilage.

Articular cartilage, obtained from the large toe during hallux valgus operations in 37 patients, was investigated for the presence of amyloid by using the Congo red staining method. Amyloid deposits were demonstrated, particularly in the superficial layer of the cartilage, in 30 cases. This amyloid did not react immunohistochemically with any of the antibodies against the known five major amyloid types (AA, A lambda, A kappa, AF, AB). From these data it is concluded that hyaline cartilage in older individuals is prone to infiltration by an amyloid of a hitherto unidentified class. From the morphological observations there seems to be no correlation between amyloid deposits and the development of osteoarthrosis.