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Bone Samples (bone + sample)
Selected AbstractsCancellous Bone Remodeling Occurs in Specialized Compartments Lined by Cells Expressing Osteoblastic MarkersJOURNAL OF BONE AND MINERAL RESEARCH, Issue 9 2001Ellen M. Hauge Abstract We describe a sinus, referred to as a bone remodeling compartment (BRC), which is intimately associated with cancellous bone remodeling. The compartment is lined on its marrow side by flattened cells and on its osseous side by the remodeling bone surface, resembling a roof of flattened cells covering the bone surface. The flat marrow lining cells are in continuity with the bone lining cells at the margins of the BRC. We examined a large number of diagnostic bone biopsy specimens received during recent years in the department. Furthermore, 10 patients (8 women and 2 men, median age 56 [40,69] years) with the high turnover disease of primary hyperparathyroidism who were treated with parathyroidectomy and followed for 3 years were included in the histomorphometric study. Bone samples for the immuno-enzyme staining were obtained from an amputated extremity of child. The total cancellous bone surface covered by BRC decreases by 50% (p < 0.05) following normalization of turnover and is paralleled by a similar 50% decrease in remodeling surface (p < 0.05). The entire eroded surface and two-thirds of the osteoid surface are covered by a BRC. BRC-covered uncompleted walls are 30% (p < 0.05) thinner than those without a BRC. This indicates that the BRC is invariably associated with the early phases of bone remodeling, that is, bone resorption, whereas it closes during the late part of bone formation. Immuno-enzyme staining shows that the flat marrow lining cells are positive for alkaline phosphatase, osteocalcin, and osteonectin, suggesting that they are bone cells. The first step in cancellous bone remodeling is thought to be the lining cells digesting the unmineralized matrix membrane followed by their disappearance and the arrival of the bone multicellular unit (BMU). We suggest that the lining cell barrier persists during bone remodeling; that the old lining cells become the marrow lining cells, allowing bone resorption and bone formation to proceed under a common roof of lining cells; that, at the end of bone formation, new bone lining cells derived from the flattened osteoblasts replace the marrow lining cells thereby closing the BRC; and that the two layers of lining cells eventually becomes a single layer. The integrity of the osteocyte-lining cell system is reestablished by the new generation of lining cells. The BRC most likely serves multiple purposes, including efficient exchange of matrix constituents and minerals, routing, monitoring, or modulating bone cell recruitment, and possibly the anatomical basis for the coupling of bone remodeling. [source] Methodological assessment of acid-etching for visualizing the osteocyte lacunar-canalicular networks using scanning electron microscopyMICROSCOPY RESEARCH AND TECHNIQUE, Issue 3 2010Daniel J. Kubek Abstract Osteocytes are the most abundant of the bone cells. Each osteocyte is contained within its own lacuna and connected to adjacent osteocytes via fillipodial processes, which form an intricate network of canaliculi within the matrix. Studying this intricate network of cells and their processes is difficult, because it exists embedded within a densely mineralized matrix. Scanning electron microscopy (SEM) has been shown to be a useful tool for visualizing this cellular network, yet the techniques involved for preparing specimens has not been systematically explored. The goal of this study was to investigate how variations in acid-etching, both etching media and etching duration, affect SEM-based visualization of the osteocyte lacunar,canalicular network. Bone samples were embedded in plastic and then acid etched in either 9% (10, 20, 40, and 60 s durations) or 37% (5, 10, and 15 s) phosphoric acid. Specimens were imaged using SEM, and qualitative evaluation of the lacunar,canalicular network was undertaken. Our findings show acid etchingwith a 9% phosphoric acid solution for 20 s provided the most favorable visualization of the osteocyte lacunar,canalicular network. Microsc. Res. Tech., 2010. © 2009 Wiley-Liss, Inc. [source] Histologic findings at augmented bone areas supplied with two different bone substitute materials combined with sinus floor liftingCLINICAL ORAL IMPLANTS RESEARCH, Issue 1 2004Report of one case Abstract: This case report is focused on the histologic findings of bone tissue supplied with two different hydroxyapatites (HAs) used for maxillary sinus floor grafting in the same patient after various healing intervals. An insufficient unilateral sinus floor grafting with Bio-Oss® biomaterial was followed by an additional grafting procedure with Algipore® biomaterial performed 4 years later. Bone samples obtained during second-stage dental implantation contained the interesting combination of Bio-Oss®, a bovine anorganic bone substitute, and Algipore®, a porous algae-derived HA, in close vicinity, yet after different healing periods. Light microscopy exhibited satisfactory osseointegration of both grafting materials. However, Bio-Oss® biomaterial showed no evidence of substantial remodeling after a healing period of 4.5 years. On the other hand, Algipore® particles demonstrated signs of remodeling by being locally resorbed and partially replaced with newly formed bone already within 6 months. Résumé Ce rapport d'un cas se concentre sur les découvertes histologiques du tissu osseux apporté par deux hydroxyapatites différents utilisés pour l'épaississement du plancher sinusal chez le même patient après différents intervalles de guérison. Un épaississement sinusal unilatéral insuffisant effectué avec le Bio-Oss® a été suivi d'un nouveau processus d'épaississement avec le Algipore® (un hydroxyapatite biologique dérivé d'algues marines calcifiées) quatre ans plus tard. Des échantillons osseux obtenus durant le placement des implants contenaient la combinaison intéressante du Bio-Oss®, un substitut osseux inorganique bovin, et du Algipore®, un hydroxyapatite d'algues, l'un à côté de l'autre, même si il y avait des périodes de guérison différentes. La microscopie optique a montré une ostéoïntégration satisfaisante des deux matériaux greffés. Cependant, le Bio-Oss® ne montrait aucune évidence d'un remodelage substantiel après une période de 4,5 années. Par contre les particules de Algipore® possèdaient des signes de remodelage tout en étant résorbées localement et partiellement remplacées par de l'os néoformé déjà après six mois. Zusammenfassung Diese Fallvorstellung präsentiert die Histologie eines Knochens, der bei einer in zwei Schritten und mit verschiedenen Hydroxylapatitprodukten durchgeführten Sinusbodenelevation entstanden ist. Die Einheilzeit der zwei Transplantate war verschieden lang. Eine einseitig durchgeführte und ungenügende Sinusbodenelevation mit dem Biomaterial Bio-Oss® wurde vier Jahre später mit einem zusätzlichen Augmentationseingriff unter Einsatz des Biomaterials Algipore® korrigiert. Bei der Implantation konnte man Knochenproben von interessanter Zusammensetzung entnehmen. Es handelte sich um einen Knochen, der während verschieden langer Zeit in engem Kontakt mit Bio-Oss®, einem anorganischen Rinderknochentransplantat, und Algipore®, einem porösen aus Algen gewonnenem Hydroxylapatit stand. Die Lichtmikroskopie zeigte eine zufriedenstellende Osseointegration beider Transplantatmaterialien. Bio-Oss® zeigte jedoch nach einer Einheilzeit von 4.5 Jahren noch keine Anzeichen einer substanziellen Remodellation. Bei den Algipore® Partikeln war dies bereits der Fall. Sie waren schon nach sechs Monaten an einigen Stellen anresorbiert, teilweise sogar mit neu gebildetem Knochen ersetzt. Resumen Este informe de un caso esta enfocado sobre los hallazgos histológicos de tejido óseo suministrado con dos diferentes hidroxiapatitas usadas para injerto del suelo del seno maxilar en el mismo paciente tras varios intervalos de cicatrización. Un insuficiente injerto del suelo del seno con biomaterial Bio-Oss® fue seguido por un procedimiento adicional de injerto con biomaterial Algipore® llevado a cabo 4 años mas tarde. Las muestras de hueso obtenidas durante la segunda fase de la implantación dental contenían una interesante combinación de Bio-Oss®, un sustituto óseo inorgánico bovino y Algipore®, una hidroxiapatita porosa derivada de algas, en próxima vecindad, aunque tras diferentes periodos de cicatrización. La microscopía óptica exhibió una osteointegración satisfactoria de ambos materiales de injerto. De todos modos, el biomaterial Bio-Oss® no mostró evidencia de un remodelado sustancial tras un periodo de cicatrización de 4.5 años. Por otro lado las partículas de Algipore® demostraron signos de remodelado al ser localmente reabsorbidas y reemplazadas parcialmente con hueso neoformado ya a los 6 meses. [source] Brief communication: Minimally invasive bone sampling method for DNA analysisAMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 4 2009Victoria E. Gibbon Abstract Obtaining a bone sample for DNA analysis has traditionally been a destructive practice, which has resulted in reluctance on behalf of curators for skeletal collections to allow invasive testing. A novel minimally invasive bone sampling method for DNA analysis is presented here. This method uses a conventional hand drill wherein the bone sample is extracted from the intercondylar fossa of the femur; it does not interfere with any known anthropometric landmarks and only leaves a small hole on the surface of the bone. The temperature of the drill is documented and it was established due to the minor increase in temperature, that this should not affect the molecular integrity of the sample. This method is easily replicated and is suitable for both human and other animal skeletal material and can be applied to rare specimens with little risk. Am J Phys Anthropol, 2009. © 2009 Wiley-Liss, Inc. [source] Genetic diversity of endangered brown bear (Ursus arctos) populations at the crossroads of Europe, Asia and AfricaDIVERSITY AND DISTRIBUTIONS, Issue 5 2009Sébastien Calvignac Abstract Aim, Middle East brown bears (Ursus arctos syriacus Hemprich and Ehrenberg, 1828) are presently on the edge of extinction. However, little is known of their genetic diversity. This study investigates that question as well as that of Middle East brown bear relationships to surrounding populations of the species. Location, Middle East region of south-western Asia. Methods, We performed DNA analyses on 27 brown bear individuals. Twenty ancient bone samples (Late Pleistocene to 20th century) from natural populations and seven present-day samples obtained from captive individuals were analysed. Results, Phylogenetic analyses of the mitochondrial sequences obtained from seven ancient specimens identify three distinct maternal clades, all unrelated to one recently described from North Africa. Brown bears from Iran exhibit striking diversity (three individuals, three haplotypes) and form a unique clade that cannot be linked to any extant one. Individuals from Syria belong to the Holarctic clade now observed in Eastern Europe, Turkey, Japan and North America. Specimens from Lebanon surprisingly appear as tightly linked to the clade of brown bears now in Western Europe. Moreover, we show that U. a. syriacus in captivity still harbour haplotypes closely linked to those found in ancient individuals. Main conclusion, This study brings important new information on the genetic diversity of brown bear populations at the crossroads of Europe, Asia and Africa. It reveals a high level of diversity in Middle East brown bears and extends the historical distribution of the Western European clade to the East. Our analyses also suggest the value of a specific breeding programme for captive populations. [source] Late-glacial remains of woolly mammoth (Mammuthus primigenius) from Shropshire, UK: stratigraphy, sedimentology and geochronology of the Condover siteGEOLOGICAL JOURNAL, Issue 4 2009J. D. Scourse Abstract In 1986 remains of an adult woolly mammoth, Mammuthus primigenius (Blumenbach), were discovered at Norton Farm Pit, Condover, south of Shrewsbury, UK. Preliminary stratigraphical investigations indicated that this individual dated to the Devensian Late-glacial Interstadial, then the first evidence for survival of mammoth in Britain following the Last Glacial Maximum. Initial radiocarbon analysis confirmed this interpretation. Subsequent excavations in 1987/1988 recovered the remains of a further three juvenile mammoth individuals. All of these remains were found in the spoil heaps of overburden (ex situ) and their true stratigraphical context had to be reconstructed from the remnants surviving in the Pit. The 1987/1988 excavations enabled stratigraphical investigation of the site and submission of samples for radiocarbon (14C) dating, including the use of ultrafiltration pretreatment for bone samples, with the aims of reconstructing the geological and palaeoenvironmental evolution of the site and the sedimentary context of the unstratified mammoth remains. These results are presented here. This investigation indicates that the woolly mammoth remains at Condover derive from a dead-ice landscape dominated by eskers, kames and kettle-hole basins, and that the sedimentary sequence in which the mammoth remains were found forms the infilling of a kettle-hole basin. The sedimentary infilling and formation of the kettle-hole basin through ice block melt-induced subsidence were syngenetic. 14C determinations indicate that basin infill was initiated prior to Greenland Interstadial 1, and probably in Greenland Stadial 2 i.e. before 14.7,ka BP and that it continued until the early Holocene, around 8,ka BP. The sedimentological and 14C data indicate that the unstratified mammoth remains can be attributed to a dark grey clayey sandy silt (Unit C1), which accumulated during the earlier part of Greenland Interstadial 1 (14 to 14.5,ka BP) within an actively subsiding slow-flowing, beaded, fluvial network characterized by channels and pools/lakes, and with relatively shallow marginal slopes. The sedimentary architecture indicates survival of the buried ice block into Greenland Interstadial 1 and final melting only towards the end of the Interstadial at ca. 12.65,ka BP. Copyright © 2009 John Wiley & Sons, Ltd. [source] Chronic acid ingestion promotes renal stone formation in rats treated with vitamin D3INTERNATIONAL JOURNAL OF UROLOGY, Issue 1 2007Naohiko Okamoto Objective: Although hypercalciuria, a well-established adverse effect of vitamin D3, can be a risk factor of renal stone formation, the risk of nephrolithiasis has not been well defined. The consumption of a diet high in acid precursors is often cited as a risk factor for the development of calcium-based kidney stones. In the present study, we investigated the effect of chronic acid ingestion on kidney stone formation in rats treated with calcitriol (1,25[OH]2 D3). Methods: Control rats (C-C), calcitriol-treated rats (C-V; three treatments of 0.5 µg of calcitriol per week) and acid-ingested (water containing 0.21 mol/L NH4Cl), calcitriol-treated (three treatments of 0.5 µg of calcitriol per week) rats (A-V) were fed in metabolic cages. After 1 month, urine, blood, kidney and bone samples were analyzed. Results: The A-V rats exhibited elevated serum calcium concentrations, urinary calcium and phosphate excretion, urinary type I collagen cross-linked N-peptide (NTx)/creatinine values, mRNA expression of osteopontin in the kidney, and renal calcium contents as well as decreased bone mineral densities, compared with the C-C and C-V rats. Urinary citrate excretion was lower and NaDC-1 mRNA expression in the kidney was higher in the A-V rats than in the C-C and C-V rats. Calcium phosphate kidney stones were found in the A-V rats. Conclusions: The ingestion of NH4Cl, an acid precursor, promotes calcium phosphate kidney stone formation in calcitriol-treated rats. The chronic intake of a diet rich in acid precursors may be a risk factor for the development of kidney stones in subjects who are being treated with calcitriol. [source] Site-Specific Deterioration of Trabecular Bone Architecture in Men and Women With Advancing AgeJOURNAL OF BONE AND MINERAL RESEARCH, Issue 12 2008Eva-Maria Lochmüller Abstract We tested the hypothesis that the age dependence of trabecular bone microstructure differs between men and women and is specific to skeletal site. Furthermore, we aimed to investigate the microstructural pattern of bone loss in aging. Microstructural properties of trabecular bone were measured in vitro in 75 men and 75 age-matched women (age, 52,99 yr) using ,CT. Trabecular bone samples were scanned at a 26-,m isotropic resolution at seven anatomical sites (i.e., distal radius, T10 and L2 vertebrae, iliac crest, femoral neck and trochanter, and calcaneus). DXA measurements were obtained at the distal radius and proximal femur and QCT was used at T12. No significant decrease in bone density or structure with age was found in men using ,CT, DXA, or QCT at any of the anatomical sites. In women, a significant age-dependent decrease in BV/TV was observed at most sites, which was strongest at the iliac crest and weakest at the distal radius. At most sites, the reduction in BV/TV was associated with an increase in structure model index, decrease in Tb.N, and an increase in Tb.Sp. Only in the calcaneus was it associated with a significant decrease in Tb.Th. In conclusion, a significant, site-specific correlation of trabecular bone microstructure with age was found in women but not in men of advanced age. The microstructural basis by which a loss of BV/TV occurs with age can vary between anatomical sites. [source] Irreversible Perforations in Vertebral Trabeculae?,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 7 2003X Banse In human cancellous bone, osteoclastic perforations resulting from normal remodeling were generally considered irreversible. In human vertebral samples, examined by backscatter electron microscopy, there was clear evidence of bridging of perforation defects by new bone formation. Hence trabecular perforations may not be irreversible. Introduction: Preservation of the trabecular bone microarchitecture is essential to maintain its load-bearing capacity and prevent fractures. However, during bone remodeling, the osteoclasts may perforate the platelike trabeculae and disconnect the structure. Large perforations (>100 ,m) are generally considered irreversible because there is no surface on which new bone can be laid down. In this work, we investigated the outcome of these perforations on human vertebral cancellous bone. Materials and Methods: Using backscatter electron microscopy, we analyzed 264 vertebral bone samples from the thoracic and lumbar spine of nine subjects (44,88 years old). Nine fields (2 × 1.5 mm) were observed on each block. Several bone structural units (BSUs) were visible on a single trabecula, illustrating a dynamic, historical aspect of bone remodeling. A bridge was defined as a single and recent BSU connecting two segments of trabeculae previously separated by osteoclastic resorption. They were counted and measured (length and breadth, ,m). Results and Conclusion: We observed 396 bridges over 2376 images. By comparison, we found only 15 microcalluses on the same material. The median length of the bridge was 165 ,m (range, 29,869 ,m); 86% being longer than 100 ,m and 35% longer than 200 ,m. Their breadth was 56 ,m (range, 6,255 ,m), but the thinnest were still in construction. Bridges were found in all nine subjects included in the study, suggesting that it is a common feature of normal vertebral bone remodeling. These observations support the hypothesis that perforation could be repaired by new bone formation. and hence, might not be systematically irreversible. [source] Cytokines, Osteoprotegerin, and RANKL In Vitro and Histomorphometric Indices of Bone Turnover in Patients With Different Bone Diseases,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 3 2003Heide Siggelkow Abstract Cytokines are supposed to play an essential role in the regulation of the bone metabolic unit. However, information on cytokine production of primary human osteoblasts from patients with metabolic bone disease is scarce, and few attempts have been made to correlate such data to histomorphometric parameters of individual patients. We investigated 11 patients with metabolic bone disease referred to our outpatient department for bone biopsy and analyzed interleukin (IL)-1, IL-6, and TNF-, protein release and gene expression in primary osteoblast cultures. Compared with four controls, five patients showed normal cytokine protein release, whereas six patients showed much higher levels of interleukin-6 (26-fold) and TNF-, (84-fold). All three cytokines were strongly correlated concerning gene expression and/or protein levels (r = 0.72,0.96). Histomorphometric analysis of the bone samples showed that eroded surface (ES/BS) as a parameter of bone resorption was significantly associated with TNF-,. In addition, RANKL gene expression was positively associated with ES/BS and osteoclast surface (Oc.S/BS). Finally, the formation parameters osteoid volume and osteoid surface were negatively associated with TNF-,. In conclusion, in an in vitro-ex vivo model of bone cells obtained from a group of 11 patients with different forms of metabolic bone disease, cytokine release in conditioned medium was significantly associated with bone resorption and bone formation, as quantified by histomorphometry. TNF-, seemed to be the more important cytokine; its effect on bone resorption could be mediated by RANKL. [source] High-Turnover Periprosthetic Bone Remodeling and Immature Bone Formation Around Loose Cemented Total Hip JointsJOURNAL OF BONE AND MINERAL RESEARCH, Issue 1 2001Michiaki Takagi Abstract Aseptic loosening and periprosthetic osteolysis are the major problems awaiting solution in total hip surgery. The clinical investigation focused on the analysis of periprosthetic bone remodeling to clarify one important key event in the cascade of periprosthetic connective tissue weakening and osteolysis around loose artificial hip joints. Twelve acetabular bone samples adjacent to granulomatous synovial-like membrane of loose hip prosthesis were retrieved at revision surgery and processed for Villanueva bone staining for morphological observation and bone histomorphometric analysis. Eight well-fixed bony samples were used as control. Although osteoclastic surface and eroded surface by osteoclasts were evident in the periprosthetic bone from loose hip joints (p = 0.003 and p = 0.027), increased osteoid/low-mineralized bone matrix (p < 0.001) and osteoid width (p < 0.001) also were significant findings in structural analysis. In addition, not only elevated mineral apposition rate (MAR; p = 0.044) but also increased mineralizing surface (p = 0.044) and bone formation rate (BFR; p = 0.002) in loose periprosthetic bones were shown in dynamic data analysis. These results were confirmed by precise morphological observation by confocal laser scanning microscopy. Active coupling of bone formation and resorption and increased osteocytes with abundant bone canalicular projections were found in combined with the presence of immature bone matrices (osteoid and low-mineralized bone areas) in periprosthetic bones from loose hip joints. These results indicated that active osteoclastic bone resorption and/or defective bone formation are coupled with monocyte/macrophage-mediated foreign body-type granuloma in the synovial-like interface membrane of loose hip joints. Thus, this unique high-turnover periprosthetic bone remodeling with bad bone quality probably is caused by the result of cellular host response combined with inappropriate cyclic mechanical loading. The fragile periprosthetic bone may contribute to hip prosthesis loosening. [source] A new method for three-dimensional skeleton graph analysis of porous media: application to trabecular bone microarchitectureJOURNAL OF MICROSCOPY, Issue 2 2000L. Pothuaud This paper introduces a new three-dimensional analysis of complex disordered porous media. Skeleton graph analysis is described and applied to trabecular bone images obtained by high resolution magnetic resonance imaging. This technique was developed bearing in mind topological considerations. The correspondence between vertices and branches of the skeleton graph and trabeculae is used in order to get local information on trabecular bone microarchitecture. In addition to real topological parameters, local structural information about trabeculae, such as length and volume distributions, are obtained. This method is applied to two sets of samples: six osteoporosis and six osteoarthritis bone samples. We demonstrate that skeleton graph analysis is a powerful technique to describe trabecular bone microarchitecture. [source] Effects of femtosecond laser irradiation on osseous tissuesLASERS IN SURGERY AND MEDICINE, Issue 3 2007B. Girard DMD Abstract Background and Objective Few studies have investigated femtosecond (fs) lasers for cutting bone tissue. Study Design/Materials and Methods A 775 nm, 1 kHz, 200 femtosecond, up to 400 µJ laser system was used to irradiate in vitro calcified cortical bone samples and bone tissue culture samples. Results The ablation threshold in cortical bone was 0.69±0.08 J/cm2 at 775 nm and 0.19±0.05 J/cm2 at 387 nm. Plasma shielding experiments determined that the ablation plume and the plasma significantly affect material removal at high repetition rates and appear to generate thermal transients in calcified tissue. Confocal analysis revealed intact enzymatic activity on the surface of cells immediately adjacent to cells removed by fs laser irradiation. Conclusions These experiments demonstrate that fs lasers used for bone tissue cutting do not appear to generate significant temperature transients to inactivate proteins and that cellular membrane integrity is disrupted for only a few cell layers. Lasers Surg. Med. 39:273,285, 2007. © 2007 Wiley-Liss, Inc. [source] Searching for DNA in museum specimens: a comparison of sources in a mammal speciesMOLECULAR ECOLOGY RESOURCES, Issue 3 2010M. CASAS-MARCE Abstract The number of genetic studies that use preserved specimens as sources of DNA has been steadily increasing during the last few years. Therefore, selecting the sources that are more likely to provide a suitable amount of DNA of enough quality to be amplified and at the minimum cost to the original specimen is an important step for future research. We have compared different types of tissue (hides vs. bones) from museum specimens of Iberian lynx and multiple alternative sources within each type (skin, footpad, footpad powder, claw, diaphysis, maxilloturbinal bone, mastoid process and canine) for DNA yield and probability of amplification of both mitochondrial and nuclear targets. Our results show that bone samples yield more and better DNA than hides, particularly from sources from skull, such as mastoid process and canines. However, claws offer an amplification success as high as bone sources, which makes them the preferred DNA source when no skeletal pieces have been preserved. Most importantly, these recommended sources can be sampled incurring minimal damage to the specimens while amplifying at a high success rate for both mitochondrial and microsatellite markers. [source] Diet and mobility in Early Medieval Bavaria: A study of carbon and nitrogen stable isotopesAMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 2 2010Susanne Hakenbeck Abstract This study investigates patterns of mobility in Early Medieval Bavaria through a combined study of diet and associated burial practice. Carbon and nitrogen isotope ratios were analyzed in human bone samples from the Late Roman cemetery of Klettham and from the Early Medieval cemeteries of Altenerding and Straubing-Bajuwarenstrasse. For dietary comparison, samples of faunal bone from one Late Roman and three Early Medieval settlement sites were also analyzed. The results indicate that the average diet was in keeping with a landlocked environment and fairly limited availability of freshwater or marine resources. The diet appears not to have changed significantly from the Late Roman to the Early Medieval period. However, in the population of Altenerding, there were significant differences in the diet of men and women, supporting a hypothesis of greater mobility among women. Furthermore, the isotopic evidence from dietary outliers is supported by "foreign" grave goods and practices, such as artificial skull modification. These results reveal the potential of carbon and nitrogen isotope analysis for questions regarding migration and mobility. Am J Phys Anthropol 143:235,249, 2010. © 2010 Wiley-Liss, Inc. [source] Brief communication: Mass spectroscopic characterization of tetracycline in the skeletal remains of an ancient population from Sudanese Nubia 350,550 CEAMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 1 2010Mark L. Nelson Abstract Histological evidence of tetracycline use has been reported in an ancient X-Group population (350,550 CE) from Sudanese Nubia (Bassett et al., 1980). When bone samples were examined by fluorescent microscopy under UV light at 490 Å yellow,green fluorophore deposition bands, similar to those produced by tetracycline, were observed, suggesting significant exposure of the population to the antibiotic. These reports were met skeptically with claims that the fluorescence was the result of postmortem taphonomic infiltration of bacteria and fungi. Herein, we report the acid extraction and mass spectroscopic characterization of the antibiotic tetracycline from these samples. The bone samples were demineralized in anhydrous hydrogen fluoride which dissolved the bone-complexed tetracycline, followed by isolation by solid phase extraction on reverse-phase media. Chemical characterization by high pressure liquid chromatography mass-spectroscopic procedures showed that the retention times and mass spectra of the bone extract were identical to tetracycline when treated similarly. These results indicate that a natural product tetracycline was detectable within the sampled bone and was converted to the acid-stable form, anhydrotetracycline, with a mass + H of 427.1 amu. Our findings show that the bone sampled is labeled by the antibiotic tetracycline, and that the NAX population ingested and were exposed to tetracycline-containing materials in their dietary regime. Am J Phys Anthropol 143:151,154, 2010. © 2010 Wiley-Liss, Inc. [source] Technical note: PCR analysis of minimum target amount of ancient DNAAMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 2 2010Daniela Woide Abstract The study of ancient DNA plays an important role in archaeological and palaeontological research as well as in pathology and forensics. Here, we present a new tool for ancient DNA analysis, which overcomes contamination problems, DNA degradation, and the negative effects of PCR inhibitors while reducing the amount of starting target material in the picogram range. Ancient bone samples from four Egyptian mummies were examined by combining laser microdissection, conventional DNA extraction, and low-volume PCR. Initially, several bone particles (osteons) in the micrometer range were extracted by laser microdissection. Subsequently, ancient DNA amplification was performed to verify our extraction method. Amelogenin and ,-actin gene specific fragments were amplified via low-volume PCR in a total reaction volume of 1 ,l. Results of microdissected mummy DNA samples were compared to mummy DNA, which was extracted using a standard DNA extraction method based on pulverization of bone material. Our results highlight the combination of laser microdissection and low-volume PCR as a promising new technique in ancient DNA analysis. Am J Phys Anthropol, 2010. © 2010 Wiley-Liss, Inc. [source] Technical note: Efficiency of total demineralization and ion-exchange column for DNA extraction from boneAMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 1 2010Seung Bum Seo Abstract We investigated whether a combination of recently introduced methods, total demineralization and ion-exchange columns, would increase DNA recovery from old bone. Ten bone samples taken after a burial period of ,60 years were used in this study. Bone powder was digested using total or incomplete demineralization. DNA was extracted by the standard organic method. The DNA extract was purified with ion-exchange columns or QIAquick® spin columns. The efficiency of different DNA extraction methods was compared in terms of DNA concentration, inhibitors generated by real-time PCR, and conventional STR typing results. The mean DNA concentration using the total demineralization method is ,3 times higher than that using the incomplete demineralization method. For DNA purification, the method using QIAquick® spin columns appeared to yield approximately double the DNA than the method using ion-exchange columns. Furthermore, 2 out of 10 samples showed higher levels of inhibition with CT values of IPC ,30 cycles when using only ion-exchange columns. In STR results, total demineralization yielded more locus profiles by 4.2 loci than incomplete demineralization, and QIAquick® spin columns also yielded more locus profiles by 3.5 loci than ion-exchange columns. Total demineralization of bone powder significantly increased DNA yield and improved STR typing results. However, the use of ion-exchange columns was not efficient when compared with the method using QIAquick® spin columns. It is suggested that the combination of total demineralization and QIAquick® spin columns lead to greatly improved STR typing results. Am J Phys Anthropol 2010. © 2009 Wiley-Liss, Inc. [source] Isotopic evidence for age-related variation in diet from Isola Sacra, ItalyAMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 1 2005Tracy L. Prowse Abstract This study examines collagen (N = 105) and apatite (N = 65) data from an Imperial Roman skeletal sample from the necropolis of Isola Sacra (Rome, Italy). This paper explores correlations between the isotopic composition of bone samples and the inferred age and sex of these individuals (aged 5,45+ years). The collagen of males, and older individuals in general, was significantly enriched in 15N but not 13C. Bone carbonate was somewhat depleted in 13C in some older individuals, suggesting increased consumption of olive oil and possibly wine. Subadults (>5 years) in the sample appear to have consumed an almost exclusively terrestrial diet. This study demonstrates a clear trend in dietary patterns between adult age groups, as well as between adults and children within a population. Am J Phys Anthropol 128:2-13, 2005. © 2005 Wiley-Liss, Inc. [source] MtDNA from extinct Tainos and the peopling of the CaribbeanANNALS OF HUMAN GENETICS, Issue 2 2001C. LALUEZA-FOX Tainos and Caribs were the inhabitants of the Caribbean when Columbus reached the Americas; both human groups became extinct soon after contact, decimated by the Spaniards and the diseases they brought. Samples belonging to pre-Columbian Taino Indians from the La Caleta site (Dominican Republic) have been analyzed, in order to ascertain the genetic affinities of these groups in relation to present-day Amerinds, and to reconstruct the genetic and demographic events that took place during the peopling of the Caribbean. Twenty-seven bone samples were extracted and analyzed for mtDNA variation. The four major Amerindian mtDNA lineages were screened through amplification of the specific marker regions and restriction enzymatic digestion, when needed. The HVRI of the control region was amplified with four sets of overlapping primers and sequenced in 19 of the samples. Both restriction enzyme and sequencing results suggest that only two (C and D) of the major mtDNA lineages were present in the sample: 18 individuals (75%) belonged to the C haplogroup, and 6 (25%) to the D haplogroup. Sequences display specific substitutions that are known to correlate with each haplogroup, a fact that helped to reject the possibility of European DNA contamination. A low rate of Taq misincorporations due to template damage was estimated from the cloning and sequencing of different PCR products of one of the samples. High frequencies of C and D haplogroups are more common in South American populations, a fact that points to that sub-continent as the homeland of the Taino ancestors, as previously suggested by linguistic and archaeological evidence. Sequence and haplogroup data show that the Tainos had a substantially reduced mtDNA diversity, which is indicative of an important founder effect during the colonization of the Caribbean Islands, assumed to have been a linear migratory movement from mainland South America following the chain configuration of the Antilles. [source] ERK-1/2 and p38 in the regulation of hypertrophic changes of normal articular cartilage chondrocytes induced by osteoarthritic subchondral osteoblastsARTHRITIS & RHEUMATISM, Issue 5 2010Indira Prasadam Objective Previous studies have shown the influence of subchondral bone osteoblasts (SBOs) on phenotypical changes of articular cartilage chondrocytes (ACCs) during the development of osteoarthritis (OA). The molecular mechanisms involved during this process remain elusive, in particular, the signal transduction pathways. The aim of this study was to investigate the in vitro effects of OA SBOs on the phenotypical changes in normal ACCs and to unveil the potential involvement of MAPK signaling pathways during this process. Methods Normal and arthritic cartilage and bone samples were collected for isolation of ACCs and SBOs. Direct and indirect coculture models were applied to study chondrocyte hypertrophy under the influence of OA SBOs. MAPKs in the regulation of the cell,cell interactions were monitored by phosphorylated antibodies and relevant inhibitors. Results OA SBOs led to increased hypertrophic gene expression and matrix calcification in ACCs by means of both direct and indirect cell,cell interactions. In this study, we demonstrated for the first time that OA SBOs suppressed p38 phosphorylation and induced ERK-1/2 signal phosphorylation in cocultured ACCs. The ERK-1/2 pathway inhibitor PD98059 significantly attenuated the hypertrophic changes induced by conditioned medium from OA SBOs, and the p38 inhibitor SB203580 resulted in the up-regulation of hypertrophic genes in ACCs. Conclusion The findings of this study suggest that the pathologic interaction of OA SBOs and ACCs is mediated via the activation of ERK-1/2 phosphorylation and deactivation of p38 phosphorylation, resulting in hypertrophic differentiation of ACCs. [source] | |||||||||||||