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Here, we describe a partial cranium of a large canid dated at 406.5 ± 2.4 ka from the Middle Pleistocene of Ponte Galeria (Rome, Italy). The sample represents one of the few Middle Pleistocene remains of a wolf-like canid falling within the timeframe when the Canis mosbachensis–Canis lupus transition occurred, a key moment to understand the spread of the extant wolf (Canis lupus) in Europe. CT-based methods allow studying the outer and inner cranial anatomy (brain and frontal sinuses) of a selected sample of fossil and extant canids. Morphological and biometric results allowed to: (I) ascribe the cranium from Ponte Galeria to an adult Canis lupus, representing the first reliable occurrence of this taxon in Europe; (II) provide the content for a biochronological revision of the Middle Pleistocene record of European wolves.
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Motivation: DnaSP is a software package for a comprehensive analysis of DNA polymorphism data. Version 5 implements a number of new features and analytical methods allowing extensive DNA polymorphism analyses on large datasets. Among other features, the newly implemented methods allow for: (i) analyses on multiple data files; (ii) haplotype phasing; (iii) analyses on insertion/deletion polymorphism data; (iv) visualizing sliding window results integrated with available genome annotations in the UCSC browser. Availability: Freely available to academic users from: http://www.ub.edu/dnasp Contact: jrozas{at}ub.edu
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A new approach to rapid sequence comparison, basic local alignment search tool (BLAST), directly approximates alignments that optimize a measure of local similarity, the maximal segment pair (MSP) score. Recent mathematical results on the stochastic properties of MSP scores allow an analysis of the performance of this method as well as the statistical significance of alignments it generates. The basic algorithm is simple and robust; it can be implemented in a number of ways and applied in a variety of contexts including straightforward DNA and protein sequence database searches, motif searches, gene identification searches, and in the analysis of multiple regions of similarity in long DNA sequences. In addition to its flexibility and tractability to mathematical analysis, BLAST is an order of magnitude faster than existing sequence comparison tools of comparable sensitivity.
Preprint
The Sicilian wolf represented the only population of wolves living on a Mediterranean island until the first half of the twentieth century (1930s-1960s) 1–7 . Previous studies hypothesised that they remained isolated from mainland wolves from the end of the Last Glacial Maximum (LGM) 8,9 , until human persecutions led them to extinction 1–7 . There are only seven known Sicilian wolf specimens from the 19th and 20th century preserved in museums in Italy and recent morphometric analyses assigned them to the new subspecies Canis lupus cristaldii ¹⁰ . To better understand the origins of the Sicilian wolf, and its relationship to other wolf populations, we sequenced four whole genomes (3.8×-11.6×) and five mitogenomes. We investigated the relationship between Sicilian wolves and other modern breeds to identify potential admixture. Furthermore, considering that the last land-bridge between Sicily and Italy disappeared after the LGM ¹¹ , around 17 kya, we explored the possibility that the Sicilian wolf retained ancestry from ancient wolf and dog lineages. Additionally, we explored whether the long-term isolation might have affected the genomic diversity, inbreeding levels and genetic load of the Sicilian wolf. Our findings show that the Sicilian wolves shared most ancestry with the modern Italian wolf population but are better modelled as admixed with European dog breeds, and shared traces of Eneolithic and Bronze age European dogs. We also find signatures of severe inbreeding and low genomic diversity at population and individual levels due to long-term isolation and drift, suggesting also low effective population size.
Article
Evidence of maternal care and childbirth events in the past are rare in the archaeological record and are difficult to recognize. To combat this, we analyzed thirteen double burials potentially related to childbirth death events, thereby containing an adult and a perinate. The specimens were excavated from the archaeological area identified as “Forlì Campus” (Forlì, Italy), that dated to 17th-18th centuries AD and was adjacent to a hospital in use at that time. This period witnessed the development of medical techniques and novel approaches in obstetrics in Europe, with the introduction of lying-in hospitals and maternity wards. We here tested if the double burials were ascribable to childbirth death events and thus represent the first reported cases of the hospitalization of childbirth in the history of medicine. A multidisciplinary analysis was undertaken to achieve this aim, combining anthropology, archaeology, paleopathology and archaeogenetics. In five burials the adult individual was recognized as a female in fertile age and the non-adult individual was assigned as perinate. Mitochondrial DNA analysis highlighted different haplotypes among the individuals of these burials, and these results, combined with the archaeological and anthropological data do not support a possible maternal relationship between them. This study is novel in testing the hypotheses of childbirth deaths, through a reliable approach in the interpretation of these archaeological contexts. The analysis of ancient DNA in this particular application proves a useful strategy to support and complete the interpretation of archaeological and anthropological data, showing that a general assumption of mother/child relations within such burials can be misleading.
Article
The phyletic relationship between Canis lupus and the Early-Middle Pleistocene Canis mosbachensis is widely accepted among scholars, although the taxonomy of several European fossil specimens is still debated. In the last decades, many studies focused on the evolution of Pleistocene wolves have been proposed considering new materials as well as specimens belonging to historical collections. The canid remains recovered during the last century and housed in museums, undoubtedly represent a valuable source of biometric and morphological data which require, in some cases, an updated review of the fossil material. Indeed, cases of stratigraphic and taxonomic misinterpretations represent a potential loss of information for understanding the palaeobiology of these iconic predators. Here we provide the first updated description of the specimen belonging to the collection of the Italian geologist and palaeontologist Alessandro Portis dating back to the early last century. Previously reported as Canis lupus, the cranium from the historical excavation of Ostiense palaeontological site (Rome, Italy) is ascribed to the Early-Middle Pleistocene wolf Canis mosbachensis. Moreover, the reassessment of the stratigraphic and geological data of the Ostiense site led to consider the studied specimen as the last occurrence of the Mosbach wolf in the Italian fossil record.
Article
The recently-developed statistical method known as the "bootstrap" can be used to place confidence intervals on phylogenies. It involves resampling points from one's own data, with replacement, to create a series of bootstrap samples of the same size as the original data. Each of these is analyzed, and the variation among the resulting estimates taken to indicate the size of the error involved in making estimates from the original data. In the case of phylogenies, it is argued that the proper method of resampling is to keep all of the original species while sampling characters with replacement, under the assumption that the characters have been independently drawn by the systematist and have evolved independently. Majority-rule consensus trees can be used to construct a phylogeny showing all of the inferred monophyletic groups that occurred in a majority of the bootstrap samples. If a group shows up 95% of the time or more, the evidence for it is taken to be statistically significant. Existing computer programs can be used to analyze different bootstrap samples by using weights on the characters, the weight of a character being how many times it was drawn in bootstrap sampling. When all characters are perfectly compatible, as envisioned by Hennig, bootstrap sampling becomes unnecessary; the bootstrap method would show significant evidence for a group if it is defined by three or more characters.
Article
Herein we describe for the rst time a canid partial cranium from the Contrada Monticelli site. Morphological and biometrical studies allow the fossil remains to be referred to the Middle Pleistocene wolf Canis mosbachensis. Associated taxa include Paleoloxodon antiquus, Stephanorhinus hundsheimensis, cervids, equids and bovids, whose biochronological occurrence allows the site to be referred to the Galerian Mammal Age. Diagnostic characters normally used to distinguish Canis mosbachensis from Canis lupus are herein discussed. These carnivorans show a wide range in body-size and morphological variability, related to an extensive geographical distribution. The analyzed fossil can be considered as the smallest European specimen referable to the Mosbach wolf and represents the southernmost occurrence of this taxon in Italy.
Article
Aim Grey wolves (Canis lupus) are widespread across the Holarctic. Here, we test the previously proposed hypothesis that extant North American wolves originate from multiple waves of colonization from Asia. We also test the hypothesis that land connections have been important in the evolutionary history of other isolated wolf populations in Japan. Location Holarctic. Methods We analyse 105 previously published and newly obtained complete mitochondrial genomes from a geographically diverse sample of grey wolves and date critical branches in the phylogenetic tree. Phylogeographical hypotheses are tested in an approximate Bayesian computation approach. Results We find that the mitogenomes of all living wolves in North America, including Mexican wolves, most likely derive from a single colonization event from Eurasia that expanded the grey wolf range into North America. This colonization occurred while a land bridge connected Eurasia and North America before the Cordillerian and Laurentide ice sheets fused in the Last Glacial Maximum, c. 23 ka, much more recent than predicted based on the fossil record. Pleistocene land bridges also facilitated the separate colonization of Hokkaido and the southern Japanese islands. Main conclusions Extant wolf lineages in North America derive from wolves that migrated into North America coincident with the formation of the most recent land bridge with Eurasia. The maternal lineages from earlier Pleistocene American wolves are not represented in living American wolves, indicating that they left no descendants. The timing of colonization of North America, Hokkaido and the southern Japanese islands corresponds to the changes in land connectivity as a consequence of changing sea level.
Book
Abundantly illustrated overview of the latest findings in tissue engineering and their clinical application Offers state-of-the-art presented by leading experts in the field Adopts a visual approach including self-explanatory graphics, diagrams, and photos Tissue engineering is a multidisciplinary field incorporating the principles of biology, chemistry, engineering, and medicine to create biological substitutes of native tissues for scientific research or clinical use. Applications of this technology include studies of tissue development and function, investigations of drug response, and tissue repair and replacement. Tissue engineering is rapidly becoming one of the most promising treatment options for patients suffering from tissue failure. This abundantly illustrated and clearly structured guide, written by leading experts, is intended to serve as a reference for all clinicians and researchers who deal with tissue engineering issues in their daily practice and for students studying tissue engineering at both the undergraduate and the graduate level. It will also be invaluable for professionals in related research areas, particularly those where cell and tissue culture is a new or emerging tool. The book is divided into three sections that present the latest findings in tissue engineering and their application in clinical practice. The first section discusses the basics and principles of tissue engineering, including scaffolds, cells, and technologies. The second and third sections then address in detail the tissue engineering of specific organs and tissue types. A visual approach is emphasized throughout, with numerous self-explanatory graphics, diagrams, and photos.
Article
A new method called the neighbor-joining method is proposed for reconstructing phylogenetic trees from evolutionary distance data. The principle of this method is to find pairs of operational taxonomic units (OTUs [= neighbors]) that minimize the total branch length at each stage of clustering of OTUs starting with a starlike tree. The branch lengths as well as the topology of a parsimonious tree can quickly be obtained by using this method. Using computer simulation, we studied the efficiency of this method in obtaining the correct unrooted tree in comparison with that of five other tree-making methods: the unweighted pair group method of analysis, Farris's method, Sattath and Tversky's method, Li's method, and Tateno et al.'s modified Farris method. The new, neighbor-joining method and Sattath and Tversky's method are shown to be generally better than the other methods.
Article
We evaluated the accuracy and precision of tooth wear for aging gray wolves (Canis lupus) from Alaska, Minnesota, and Ontario based on 47 known-age or known-minimum-age skulls. Estimates of age using tooth wear and a commercial cementum annuli-aging service were useful for wolves up to 14 years old. The precision of estimates from cementum annuli was greater than estimates from tooth wear, but tooth wear estimates are more applicable in the field. We tended to overestimate age by 1-2 years and occasionally by 3 or 4 years. The commercial service aged young wolves with cementum annuli to within ±1 year of actual age, but under estimated ages of wolves ≥9 years old by 1-3 years. No differences were detected in tooth wear patterns for wild wolves from Alaska, Minnesota, and Ontario, nor between captive and wild wolves. Tooth wear was not appropriate for aging wolves with an underbite that prevented normal wear or severely broken and missing teeth.
Article
Skeletal pathologies and oral disease are still largely unexplored in fossil carnivores, thus types and course of pathologies in these fossil animals are poorly known. Dental abnormalities, fractures, trauma, supernumerary teeth, tumours, periodontitis, and other bacterial infections are some of the diseases that leave traces on fossilized skulls, but their identification is not always possible by external observation on the specimen. Moreover a large number of pathologies are "hidden", partially or completely invisible on the external surface of the bones because their development took place within the bones. The degree and the type of fossilization, the state of preservation and the fossil size are just a few factors that influence the analysis of these structures. Digital viewing is a very useful tool to solve such difficulties. X-ray study can provide valuable information on bone and teeth diseases, by allowing the "visualization" of the internal structure of the bones, without the alteration and/or destruction of the specimen. Many aspects of the life of carnivores are regulated by the health condition, and in particular by the teeth and jaws condition, therefore individuals with evident disability due to the pathology and injuries are not able to perform properly some basic activities, such as foraging and defence. This paper presents new methods of non- invasive analysis in order to identify and understand these pathologies, to perform much more detailed palaeoecological reconstructions of extinct carnivores.
Article
In recent years, the application of three dimensional tomographic images in vertebrate palaeontology has contributed to improve and greatly expand the range of information derived from the study of fossilized bones. By using computed tomography (CT) scan and medical software it is possible to procure precise measurements, analyses of the internal structures and the material density gradient, carry out virtual restoration, with bones and teeth separated from lithological matrix, produce virtual casts of cavities (frontal sinuses, brains, inner ear) and hence provide greater definition in the search for diagnostic elements. It is therefore possible to obtain important information from the virtual models, that otherwise could not be acquired using conventional investigation techniques. Due to the versatility of these technologies, application of this kind of analysis is becoming crucial in many sectors of vertebrate palaeontology, especially in palaeoneurology, palaeopathology and in 3D reconstructions.
Article
The brown bear Ursus arctos, wolf Canis lupus, and Eurasian lynx Lynx lynx vanished during the 18th and 19th centuries from all regions of high human activity in Europe because of direct persecution and environmental changes. Bear, wolf, and lynx were vulnerable in different ways to deforestation and the destruction of wild ungulate populations. Analysing the ecological factors responsible for the fall of the large carnivores can help to prepare their recovery. The return of large predators into semi-natural areas such as the Alps is possible, as the forests have expanded, and the wild ungulate populations increased. Lynx reintroduction in the Alps started in the 1970s. Wolves returned to the south-western Alps from the central Italian population in the early 1990s. The brown bear is recolonising the Austrian Alps from Slovenia. However, the modern protective legislation is not backed by a cooperative attitude among the affected people. In rural areas, large carnivores are still regarded as unrestrained killers of wildlife and livestock. Ecological conditions and husbandry in the Alps have been altered substantially since the large carnivores were eradicated, and the potential for conflicts has diminished. But stockmen have lost any remaining tradition of coexistence with large predators, and sheep are again very abundant in the Swiss Alps. The return of the large predators will not be possible without changing the system of sheep-husbandry. The rural people are not yet willing to do so. They generally object to any change in their lifestyle induced from outside, and the large predators become a negative symbol for restrictive conservation measures considered to hinder economic development. Nature conservation, including the reintegration of large predators, must be integrated into rural development; local people must be much more involved in this process.
Article
In the sub-fossil assemblages of Europe the red fox is clearly the most frequent carnivorous mammalian species with a total of 1553 records. In depositions from the Weichselian Glacial the red fox Vulpes vulpes is, a typical representative of the Holocene fauna, already recorded in 100 assemblages. The Iberian peninsula, Italian peninsula and Balkans were theorised as glacial refugia. Well-founded facts give reason to believe that V vulpes was also distributed in the Carpathian refuge. Later on, the Crimean peninsula would also appear to be a possible glacial refuge of the red fox. In the last warmer complex of interstadials during the Pleni-Glacial (Hengelo-Denekamp, 38,000-25,000 BC) the red fox was distributed in central Europe. Its distribution during this epoch extended at least in part to southern England. The earliest well-dated records of V vulpes in central Europe after the Maximum Glaciation lie between 14,000 and 13,500 BC. Already during the early Late-Glacial (13,500 BC) the red fox appeared in typical glacial faunal communities. A separation to glacial refugia was only possible for 10,000 years. During the last warmer Pleni-Glacial complex of interstadials (38,000-25,000 BC) in central Europe a sympatric distribution of the arctic fox (Alopex tagopus) and the red fox probably existed. During the Last Glacial Maximum (22,000-18,000 BC) the arctic fox was exclusively distributed in central Europe, outside of the refuges. The combined distribution of A. lagopus and V vulpes during the Late-Glacial (15,000-9500 BC) in central Europe, with the probable exception of the Allerod, is precisely documented by sub-fossil assemblages. In the Pleni-Glacial the wolf Canis lupus was distributed in geographic regions that served as glacial refugia of more warm-climate adapted species. Concerning the wolf no drastic decrease of the distribution is assumed. The Holocene presence of C. lupus is probably not caused by recolonisation.
Article
The wolf is thought to have been abundant in many parts of medieval Europe, but its remains are rarely identified in archaeological contexts. One of the potential reasons for this is the problem of distinguishing between the skeletal elements of wolves and dogs, accentuated by poor preservation and fragmentation. This paper reviews the extent of this problem, exploring the morphological relationships between wolves and dogs, as well as the issue of hybridisation, and goes on to suggest how the scarcity of wolf remains may in fact reflect infrequent hunting. This is illustrated with a comparative regional case study of wolf hunting and commercial exploitation in medieval England and southern Scandinavia, synthesising archaeological and written sources. The paper concludes with an optimistic appraisal of the value of wolf remains in medieval archaeological contexts for a broader understanding of relations between humans and wolves in the medieval period. Copyright © 2006 John Wiley & Sons, Ltd.
Article
The role of vascular endothelial cells (EC) in periodontitis was investigated in a series of histological studies. Expansion of the vasculature was found to occur with development of gingivitis and periodontitis. This was thought to contribute to the characteristic tissue degradation in the developing disease. Vascular expansion could also play a role in the formation of a previously unreported perivascular hyaline material (PHyM). Polymorphonuclear leukocytes (PMN) are known to be protective in periodontitis, and the location, incidence and extent of PHyM suggested a role for PHyM in periodontitis by inhibiting PMN emigration. PMN emigration was found to occur from specialized high EC (HEC) lined post capillary venules. This was unexpected, as such vessels have previously been found to exchange lymphocytes almost exclusively. Detailed histochemical, ultrastructural and biosynthetic studies of these specialized blood vessels led to the suggestion that HEC may be specially adapted for the synthesis of cytokines in periodontitis. A negative association between expression of the membrane bound ectoenzyme, alkaline phosphatase, and HEC suggested a role for this enzyme in leukocyte emigration. These observations compel re-evaluation of the role of EC in chronic inflammation, and in periodontitis in particular. The direction of current and future work is discussed.
Article
Caninae is one of the most studied mammalian groups, nevertheless there are relatively few comparative studies on their neuroanatomy. This work contributes to a better knowledge of this subfamily, since it describes the external cerebrum anatomy of 29 out of the 35 living Caninae species, 11 of which are described for the first time. Information about their frontal region appears to be a welcome supplement to the study of the phylogeny. Two distinctive features are recognized, that can be traced back in the fossil record: the sulcal pattern medial to the coronal sulci, and the shape and relative size of the proreal gyrus. Four types are described for the first feature: (1) orthogonal: Canis, Lycaon, Cuon, Atelocynus, Speothos, (2) pentagonal: Vulpes, Alopex, Otocyon,†Eucyon, (3) parenthesis-like: †Dusicyon, Pseudalopex, Chrysocyon, (4) heart-shaped: Urocyon, Cerdocyon, Pseudalopex culpaeus, Nyctereutes. Three types are described for the second feature: (1) elongated and bilaterally compressed: Canis, Cuon, Lycaon, Atelocynus, Speothos, Cerdocyon,†Dusicyon, Chrysocyon, Pseudalopex, †Nyctereutes sinensis, †N. tingi, (2) small: Vulpes, Otocyon, Urocyon, Alopex, (3) wide and low: Nyctereutes procyonoides. On the basis of these features some phylogenetic interpretations are presented: the fossil Asian Nyctereutes is close to Cerdocyon, Speothos is close to Atelocynus, Chrysocyon is not related to Canis, Urocyon differs from Vulpes and Pseudalopex culpaeus differs from the rest of the Pseudalopex species. © 2003 The Linnean Society of London. Zoological Journal of the Linnean Society, 2003, 138, 505–522.
Article
Conflicting interpretations of the influence of coyote hybridization on wolf recovery in the western Great Lakes (WGL) states have stemmed from disagreement over the systematics of North American wolves. Questions regarding their recovery status have resulted. We addressed these issues with phylogenetic and admixture analysis of DNA profiles of western wolves, WGL states wolves and Wisconsin coyotes developed from autosome and Y-chromosome microsatellites and mitochondrial DNA control region sequence. Hybridization was assessed by comparing the haplotypes exhibited by sympatric wolves and coyotes. Genetic variability and connectivity were also examined. These analyses support the recognition of Canis lycaon as a unique species of North American wolf present in the WGL states and found evidence of hybridization between C. lupus and C. lycaon but no evidence of recent hybridization with sympatric coyotes. The recolonized WGL states wolves are genetically similar to historical wolves from the region and should be considered restored. KeywordsEndangered species recovery-mtDNA-Y-chromosome-Autosomal microsatellites-Genetic diversity-Admixture analysis-Hybridization- Canis latrans - Canis lupus - Canis lycaon
Article
The purpose of this study was to examine the regeneration of periodontal tissue after the application of recombinant human bone morphogenetic protein-2 (rhBMP-2) to horizontal circumferential defects created by experimental periodontitis. Twelve mandibular second premolars in 6 adult beagle dogs were subjected to experimental periodontal breakdown by placing silk ligatures around the teeth until the bone loss exceeded half of the root length. Flap surgery was then performed and the exposed cementum removed. The distance between the bone crest and cemento-enamel junction (CEJ) was about 5 mm. RhBMP-2 (40 micrograms/100 microliters) with a sponge-type carrier material made of gelatin and polylactic acid polyglycolic acid copolymer was placed in the furcation area (5 mm x 5 mm x 5 mm) and around the roots (10 mm x 5 mm x 2.5 mm x 2 pieces). In the control group, the same carrier material without rhBMP-2 was placed in the same manner. The flaps were replaced and sutured to cover these materials completely. Twelve weeks after surgery, the animals were sacrificed and serial sections were prepared in a bucco-lingual plane. Considerable new bone formation was observed in the rhBMP-2-treated sites. New cementum with Sharpey's fibers was observed on the instrumented root surface. On histometric analysis, the amount of new bone, new cementum, and connective tissue attachment was significantly greater in the rhBMP-2-treated group (paired t test; P < 0.01). These results indicate that suitable application of rhBMP-2 can produce considerable periodontal tissue regeneration, even in cases of horizontal circumferential defects.