Palaeopathology of human remains from the Roman Imperial Age

The increasing attention of archaeological and anthropological research towards palaeopathological studies has allowed to focus the examination of many skeletal samples on this aspect and to evaluate the presence of many diseases afflicting ancient populations. This paper describes the most interesting diseases observed in skeletal samples from five necropolises found in urban and suburban areas of Rome during archaeological excavations in the last decades, and dating back to the Imperial Age. The diseases observed were grouped into the following categories: articular diseases, traumas, infections, metabolic or nutritional diseases, congenital diseases and tumors, and some examples are reported for each group. Although extensive epidemiological investigation in ancient skeletal records is impossible, palaeopathology allowed highlighting the spread of numerous illnesses, many of which can be related to the life and health conditions of the Roman population.     

Weaning initiates a rapid and powerful anabolic phase in the rat maternal skeleton

Maternal skeletal mineral lost during lactation is rapidly restored after weaning. The purposes of this study were to determine when increases of bone formation occur after weaning, whether the expanding osteoblast population is derived from proliferating progenitors, and to relate these skeletal changes to known endocrine events at weaning. Female rats were allowed to complete one reproductive cycle. Half of these rats were mated a second time and allowed to lactate for 20 days. The other half served as an age-matched, normal estrus cycling comparison group. One day after weaning, the dams and their comparison group were given four injections of bromodeoxyuridine (BrdU) at 8-h intervals. Indices of bone formation and the kinetics of BrdU-labeled cells were measured in lumbar vertebral cancellous bone. At 2 days after weaning, cancellous bone formation rates were substantially greater than those in the nonmated rats. Indices of bone formation more than doubled from the second to seventh day after weaning. At 25 h after the first BrdU injection in the postweaned rats, considerable numbers of labeled cells were observed on or near the bone surface, with about 17% of the osteoblast population labeled. Labeled osteoblasts peaked at 20%-24% compared with 4% in the normal estrus cycling group. Immediately following weaning, there is a profound increase in the osteoblast population in maternal cancellous bone. Many, if not most of these newly formed osteoblasts were derived from proliferating progenitors. It is possible that the endocrine milieu of lactation expands or primes the osteoprogenitor pool for this rapid anabolic phase.     

Pathological lesions attributable to vitamin deficiency in skeletal remains from Puy St. Pierre (Briançon, France)

The aim of the present study was to evaluate the presence of pathological alterations attributable to vitamin deficiency in osteological remains from the church of St. Pierre in Puy de St. Pierre, Briançon, France. We recorded alterations referable to scurvy, an acquired osteodystrophy due to vitamin C deficiency, which causes both skeletal and vascular lesions in humans. Scurvy presents a different clinical picture in children and adults: in the latter, in leaves skeletal signs on the palate and teeth. The recording of osteodystrophies attributable to vitamin deficiency allowed us to make general inferences about the nutritional profile of the group under study. The study of this pathology, as well as that of nutritional stresses, can be related to the geographical-environmental conditions of the population to which the sample belongs. This is important from the anthropological point of view, especially in order to examine alimentary habits within the context of the man-environments relationship.     

Horse heart acylphosphatase: purification and characterization

The use of an affinity chromatography step performed with an immunoadsorbent consisting of anti-horse muscle acylphosphatase antibodies covalently linked to Sepharose 4B allowed us to purify horse heart acylphosphatase in a very rapid and efficient fashion. As in skeletal muscle, also in heart the enzyme is present as both a mixed disulfide with glutathione and a S-S dimer. The abundance of these forms in heart is quite lower than in skeletal muscle. The comparison of the molecular forms so purified with those obtained from horse skeletal muscle showed the same aminoacid composition, tryptic fingerprint, together with strictly similar apparent molecular weight and main kinetic parameters, supporting the conclusion that the acylphosphatase present in heart is the same enzyme as that purified from skeletal muscle.     

A Seated Burial and Associated Boatstone from Northwestern Kansas

Abstract

The skeletal remains of an adult male individual from the 14TO301 site was analyzed to the extent allowed by the condition of the bony skeleton. In addition to the skeleton, one cultural object, a boatstone, is described and utilized in order to provide a rough date for the skeletal material. The cultural material is identified as Woodland and is compared w1th other Woodland material in the region. The results of the skeletal analysis compare favorably with other single male variables in the same or near region of roughly the same time period. The material also however, corresponds favorably with earlier archaic material from Colorado. The analysis of the skeletal and cultural material expands our knowledge of the distribution of the Woodland period in the high plains. Non-metric analysis is included in the analysis of the skeletal material.     

Evaluation of methods for the determination of mitochondrial respiratory chain enzyme activities in human skeletal muscle samples

The quantification of mitochondrial enzyme activities in skeletal muscle samples of patients suspected of having mitochondrial myopathies is problematic. Therefore, we have evaluated different methods for the determination of activities cytochrome c oxidase and NADH:CoQ oxidoreductase in human skeletal muscle samples. The measurement of cytochrome c oxidase activity in the presence of 200 microM ferrocytochrome c and the detection of NADH:CoQ oxidoreductase as rotenone-sensitive NADH:CoQ(1) reductase resulted in comparable citrate synthase-normalized respiratory chain enzyme activities of both isolated mitochondria and homogenates from control human skeletal muscle samples. These methods allowed the precise detection of deficiencies of respiratory chain enzymes in skeletal muscle of two patients harboring only 20 and 27% of deleted mitochondrial DNA, respectively. Therefore, citrate synthase-normalized respiratory chain activities can serve as stable reference values for the determination of a putative mitochondrial defect in human skeletal muscle.     

Muscle-derived stem cells isolated as non-adherent population give rise to cardiac, skeletal muscle and neural lineages

Stem cells with the ability to differentiate in specialized cell types can be extracted from a wide array of adult tissues including skeletal muscle. Here we have analyzed a population of cells isolated from skeletal muscle on the basis of their poor adherence on uncoated or collagen-coated dishes that show multi-lineage differentiation in vitro. When analysed under proliferative conditions, these cells express stem cell surface markers Sca-1 (65%) and Bcrp-1 (80%) but also MyoD (15%), Neuronal beta III-tubulin (25%), GFAP (30%) or Nkx2.5 (1%). Although capable of growing as non-attached spheres for months, when given an appropriate matrix, these cells adhere giving rise to skeletal muscle, neuronal and cardiac muscle cell lineages. A similar cell population could not be isolated from either bone marrow or cardiac tissue suggesting their specificity to skeletal muscle. When injected into damaged muscle, these non-adherent muscle-derived cells are retrieved expressing Pax7, in a sublaminar position characterizing satellite cells and participate in forming new myofibers. These data show that a non-adherent stem cell population can be specifically isolated and expanded from skeletal muscle and upon attachment to a matrix spontaneously differentiate into muscle, cardiac and neuronal lineages in vitro. Although competing with resident satellite cells, these cells are shown to significantly contribute to repair of injured muscle in vivo supporting that a similar muscle-derived non-adherent cell population from human muscle may be useful in treatment of neuromuscular disorders.     

A palaeobiological examination of the geological evidence for recurring outbreaks of the crown-of-thorns starfish,Acanthaster planci (L.)

Geological investigations of the crown-of-thorns starfish (COTS) have concluded that outbreaks are not only recent but also have occurred in the past. The evidence lies in the abundance of COTS skeletal elements found both on the sea floor and within the underlying sedimentary record. These studies are flawed in three respects. First, the processes of fossil preservation from the living population to the fossil assemblage have been ignored. Second, it has not been demonstrated that the fossil skeletal elements representing alleged outbreak populations of starfish are of the same age. Third, the existence of a relationship between the number of COTS skeletal elements sampled from the sedimentary record and the relative abundance of COTS in the once living population has not been substantiated. The limitations introduced when studying the fossil record need to be established through taphonomic analyses of the COTS. Techniques which will allow greater temporal resolution of skeletal element age include amino-acid geochronology, analysis of sedimentation mode and rate, and correlation among sub-surface cores. In order to establish a relationship between the number of fossil COTS elements and the original population size, methods must be developed which will relate the number of fossil skeletal elements to the relative abundance of starfish in both the fossil and death assemblages and then to relate the latter to the relative size of the original population. When these approaches are used together it may be possible to make some estimate of relative COTS abundances based on data contained in the fossil record.     

Comparative Proteomic Profiling of Dystroglycan-Associated Proteins in Wild Type, mdx, and Galgt2 Transgenic Mouse Skeletal Muscle

Dystroglycan is a major cell surface glycoprotein receptor for the extracellular matrix in skeletal muscle. Defects in dystroglycan glycosylation cause muscular dystrophy and alterations in dystroglycan glycosylation can impact extracellular matrix binding. Here we describe an immunoprecipitation technique that allows isolation of beta dystroglycan with members of the dystrophin-associated protein complex (DAPC) from detergent-solubilized skeletal muscle. Immunoprecipitation, coupled with shotgun proteomics, has allowed us to identify new dystroglycan-associated proteins and define changed associations that occur within the DAPC in dystrophic skeletal muscles. In addition, we describe changes that result from overexpression of Galgt2, a normally synaptic muscle glycosyltransferase that can modify alpha dystroglycan and inhibit the development of muscular dystrophy when it is overexpressed. These studies identify new dystroglycan-associated proteins that may participate in dystroglycan's roles, both positive and negative, in muscular dystrophy.     

Hollow structures in radiolarian skeletons

Spines are the major element of radiolarian skeletons; they are usually solid or, rarely, hollow. Subcylindrical hollow spines and other hollow structures of radiolarian skeletons appeared in the Middle Cambrian. Later, hollow skeletal structures repeatedly developed in the evolution of all radiolarian classes of the subphylum Polycystina throughout the Phanerozoic to the Recent. The analysis of appearance and distribuion in the Phanerozoic of morphotypes with hollow spines has shown that the formation of hollow skeletal structures allowed better adaptation for the planktonic mode of life and could become a key adaptation in struggle for life under stressful conditions.     

New anthropological data from Cussac Cave (Gravettian,Dordogne, France): In situ and virtual analyses of Locus 3

Cussac Cave presents a unique combination of parietal art and several hundred parts of scattered human remains, dated to the Middle Gravettian (29–28,000 cal BP). The cave is protected as a National Heritage site. As a result, only noninvasive bioanthropological analyses are allowed, consisting of in situ observations and the study of 3D models obtained by photogrammetry. Here we present the first results of these analyses of the human remains from Locus 3. Only 65 of the 106 human skeletal fragments and bones could be firmly identified. Virtual analyses were carried out on 3D models of 16 skeletal elements so that osteometric data could be provided. Despite the limitations inherent in studying commingled remains and those specific to Cussac Cave, the search for virtual pair-matching, articular congruence, and osteometric sorting allowed the allocation of twelve bones to three individuals, one late adolescent and two adults.     

Immunocharacterization of actin and its immunofluorescent localization during the developmental gametophytic stages of Allomyces arbuscula

Highly specific polyclonal antibodies against actin from Allomyces arbuscula were produced in rabbits, immunopurified by immunoblotting and specified with actin isolated from Neurospora crassa and mouse skeletal muscle. Used as immunofluorescence probes, they allowed localization of actin in the sequential gametophytic stages of the mould.     

Generation of Myospheres From hESCs by Epigenetic Reprogramming

Generation of a homogeneous and abundant population of skeletal muscle cells from human embryonic stem cells (hESCs) is a requirement for cell-based therapies and for a "disease in a dish" model of human neuromuscular diseases. Major hurdles, such as low abundance and heterogeneity of the population of interest, as well as a lack of protocols for the formation of three-dimensional contractile structures, have limited the applications of stem cells for neuromuscular disorders. We have designed a protocol that overcomes these limits by ectopic introduction of defined factors in hESCs - the muscle determination factor MyoD and SWI/SNF chromatin remodeling complex component BAF60C - that are able to reprogram hESCs into skeletal muscle cells. Here we describe the protocol established to generate hESC-derived myoblasts and promote their clustering into tridimensional miniaturized structures (myospheres) that functionally mimic miniaturized skeletal muscles⁷.     

Effects of fluoride emissions from a modern primary aluminum smelter on a local population of white-tailed deer (Odocoileus virginianus)

The influence of fluoride emissions from a modern aluminum smelter on concentrations of skeletal fluoride and dental fluorosis in a resident population of white-tailed deer was studied. The smelter was located on Mount Holly Plantation in South Carolina, and concentrations of skeletal fluoride in the deer collected at Mount Holly increased approximately five-fold 3 yr after the operation began. Increases in skeletal fluoride of less than two-fold were observed in deer obtained from Medway Plantation which has its nearest boundary 1.6 km from the smelter site. No dental fluorosis was observed in deer collected at Medway Plantation, but mild dental fluorosis was observed in a significant number of deer collected at Mount Holly Plantation. The dental fluorosis that was observed was not associated with incisor wear or with fluoride-induced molar wear. Osteofluorosis of mandibles or metacarpals was not observed in any of the deer obtained from either plantation. The data obtained from this study indicated that the presence of a modern aluminum smelter caused a detectable increase in concentration of skeletal fluoride in the resident population of white-tailed deer, but that no adverse health effects were seen.     

Isolation of a slowly adhering cell fraction containing stem cells from murine skeletal muscle by the preplate technique

This protocol details a procedure, known as the modified preplate technique, which is currently used in our laboratory to isolate muscle cells on the basis of selective adhesion to collagen-coated tissue culture plates. By employing this technique to murine skeletal muscle, we have been able to isolate a rapidly adhering cell (RAC) fraction within the earlier stages of the process, whereas a slowly adhering cell (SAC) fraction containing muscle-derived stem cells is obtained from the later stages of the process. This protocol outlines the methods and materials needed to isolate RAC and SAC populations from murine skeletal muscle. The procedure involves mechanical and enzymatic digestion of skeletal muscle tissue with collagenase XI, dispase and trypsin followed by plating the resultant muscle slurry on collagen type I-coated flasks where the cells adhere at different rates. The entire preplate technique requires 5 d to obtain the final preplate SAC population. Two to three additional days are usually required before this population is properly established. We also detail additional methodologies designed to further enrich the resultant cell population by continuing the modified preplating process on the SAC population. This process is known as replating and requires further time.     

Modeling the prehistoric Maori population

Skeletal and comparative evidence of mortality is combined with fertility estimates for the precontact Maori population of New Zealand to determine the implied rate of precontact population growth. This rate is found to be too low to populate New Zealand within the time constraints of its prehistoric sequence, the probable founding population size, and the probable population size at contact. Rates of growth necessary to populate New Zealand within the accepted time span are calculated. The differences between this minimum necessary rate and the skeletally derived rate are too large to result solely from inadequacies in the primary data. Four alternative explanations of this conundrum are proposed: 1) skeletal evidence of precontact mortality is highly inaccurate; 2) skeletal evidence of fertility is severely underestimating actual levels; 3) there was very rapid population growth in the earliest part of the sequence up to 1150 A.D., from which no skeletal evidence currently is available; or 4) the prehistoric sequence of New Zealand may have been longer than the generally accepted 1,000-1,200 years. These alternatives are examined, and a combination of the last two is found to be the most probable. The implications of this model for New Zealand prehistory and Oceanic paleodemography are discussed.